Macrophage signatures for the diagnosis and treatment of lymphoma

Abstract

The present invention provides diagnostic methods, treatment methods, and compositions for the treatment of lymphoma (e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). The present invention is at least in part based on the discovery that macrophage biomarkers are useful in methods for identifying, diagnosing, or predicting the therapeutic efficacy of treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

Description

Cross-reference of related applications

[0001] This application claims the interests of U.S. Provisional Patent Application No. 63 / 507,091, filed on 8 June 2023, the contents of which are incorporated herein by reference in their entirety.

[0002] Reference to electronic sequence listings The contents of the electronic sequence listing (146392066940seqlist.xml, size: 41,725 ​​bytes, created June 4, 2024) are incorporated herein by reference in their entirety. [Technical Field]

[0003] This disclosure relates to a method for treating B-cell proliferative disorders, such as diffuse large B-cell lymphoma (DLBCL), by administering an immunoconjugate comprising an anti-CD79b immunoconjugate in combination with an anti-CD20 antibody, one or more chemotherapeutic agents, and a corticosteroid. Related assays and kits are also provided. [Background technology]

[0004] Non-Hodgkin lymphoma (NHL) is the most common hematological malignancy in the world and the 13th most common cancer overall (Bray et al., (2018) CA Cancer J Clin, 68:394-424). Diffuse large B-cell lymphoma (DLBCL) is a malignant subtype of NHL, accounting for approximately 32.5% of all NHL cases. Patients with DLBCL present with rapidly expanding masses and are often accompanied by local and systemic symptoms such as fever, recurrent night sweats, and / or weight loss. Approximately 45%–60% of patients develop advanced disease (Ann Arbor stage III or IV). The incidence of DLBCL increases with age, with a median age of onset of 64 years (Armitage and Weisenburger, J Clin Oncol (1998) 6:2780-95). If left untreated, DLBCL patients have a median survival of approximately 6 months.

[0005] Rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) was established as the standard of care (SoC) for DLBCL more than 20 years ago. While first-line treatment for DLBCL is potentially curative, many patients either do not respond or eventually relapse. Approaches to improve current SoC therapies for DLBCL have been largely unsuccessful. This includes experimental measures such as attempts to maximize the dose density of R-CHOP (Cunningham et al., Lancet (2013) 381:1817-26, Delarue et al., Lancet Oncol (2013) 14:525-33), as well as those tested in large-scale studies in DLBCL, including BO21005 / GOYA (Vitolo et al., Blood (2016) 128:470), DA-EPOCH-R (Wilson et al., Blood (2016) 128:469), and REMARC (Thieblemont et al., Blood (2016) 128:471). Overall, as R-CHOP has been established as the SoC therapy for DLBLC, 11 randomized phase III trials have shown no benefit compared to R-CHOP in first-line treatment of DLBCL.

[0006] One recently developed alternative to R-CHOP is polatuzumab vedotin + rituximab, cyclophosphamide, doxorubicin, and prednisone (Pola-R-CHP). Pola-R-CHP was approved by the FDA in April 2023 based on the results of the POLARIX clinical trial (clinical trial ID number NCT03274492). POLARIX was a randomized, double-blind, placebo-controlled trial comparing treatment with Pola-R-CHP or R-CHOP in previously untreated DLBCL patients. In this trial, patients treated with Pola-R-CHP showed significantly longer progression-free survival (PFS) and significantly improved modified event-free survival. However, the Pola-R-CHP treatment group did not show a significant difference in complete response rate or overall survival (FDA. "FDA approves polatuzumab vedotin-piiq for previously untreated diffuse large B-cell lymphoma (not otherwise specified) and high-grade B-cell lymphoma" (2023)).

[0007] When using Pola-R-CHP as a newly available alternative to SoC therapy, there is a need in the art to better adjust the treatment of DLBCL patients, such as those with previously untreated DLBCL.

[0008] All references cited herein, including patent applications and publications, are incorporated herein by reference in their entirety. [Overview of the project]

[0009] The present invention relates to the use of macrophage biomarkers in methods for identifying, diagnosing, or predicting the efficacy of treating lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) with diagnostic methods, treatment methods, and compositions for treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

[0010] In some embodiments, the present invention features a method for identifying, diagnosing, and / or predicting whether a patient with diffuse large B-cell lymphoma (DLBCL) can benefit from treatment comprising an immunoconjugate and an anti-CD20 antibody. The method may include measuring macrophage biomarkers in a sample from the patient, and by a macrophage biomarker amount or level in the sample being below a reference macrophage biomarker amount or level, the patient is identified, diagnosed, and / or predicted to benefit from treatment comprising an immunoconjugate and an anti-CD20 antibody, wherein the immunoconjugate is given by the following formula: [ka] (In the formula, Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

[0011] In some embodiments, the present invention features a method for selecting a therapy for a patient having DLBCL. The method may include measuring macrophage biomarkers in a sample from the patient, identifying the patient as one who can benefit from a treatment comprising an immunoconjugate and an anti-CD20 antibody if the amount or level of the macrophage biomarker in the sample is below a reference amount or level of macrophage biomarker, and the immunoconjugate is given by the following formula: [ka] (In the formula, Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

[0012] In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the amount or level of a macrophage biomarker from a patient is below the amount or level of a reference macrophage biomarker, and the method further includes administering an effective amount of immunoconjugate and an effective amount of anti-CD20 antibody to the patient.

[0013] In some embodiments, the present invention is characterized by a method for treating a patient having DLBCL. The method may include (a) measuring a macrophage biomarker in a sample from the patient such that the amount or level of the macrophage biomarker in the sample is below a reference amount or level of macrophage biomarker, and (b) administering to the patient an effective amount of immunoconjugate and an effective amount of anti-CD20 antibody based on the macrophage biomarker measured in step (a), wherein the immunoconjugate has the following formula: [ka] (In the formula, Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

[0014] In some embodiments, the present invention features a method for treating a patient having DLBCL. The method may include administering an effective amount of immunoconjugate and an effective amount of anti-CD20 antibody to the patient, wherein prior to treatment, it has been determined that the amount or level of macrophage biomarkers in a sample from the patient is below a reference amount or level of macrophage biomarkers, and the immunoconjugate is of the following formula: [ka] (In the formula, Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

[0015] In some embodiments, the present invention relates to a method for treating a patient having DLBCL and in which the amount or level of macrophage biomarkers in a patient sample is below a reference amount or level of macrophage biomarkers, comprising administering to the patient an effective amount of an immunoconjugate and an effective amount of anti-CD20 antibody, wherein the immunoconjugate has the following formula: [ka] The method is characterized by comprising (wherein Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 12, and p is 1 to 8).

[0016] In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the patient is a human patient.

[0017] In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the reference macrophage biomarker amount or level is a pre-assigned macrophage biomarker amount or level. In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the reference macrophage biomarker amount or level is the amount or level of macrophage biomarkers in a reference population. In some embodiments of the method, the amount or level of macrophage biomarkers in a reference population is the median amount or level of macrophage biomarkers in the reference population.

[0018] In some embodiments of the method, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 25th percentile of the reference population. In some embodiments of the method, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 50th percentile of the reference population. In some embodiments of the method, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 75th percentile of the reference population.

[0019] In some embodiments of the method, the reference population is a population of patients with DLBCL. In some embodiments of the method, the population of patients with DLBCL has been previously treated with an immunoconjugate and an anti-CD20 antibody. In some embodiments of the method, the population of patients with DLBCL has been previously treated with an anti-CD20 antibody. In some embodiments of the method, the reference macrophage biomarker amount or level is the amount or level of macrophage biomarkers in the reference population before treatment with the immunoconjugate and anti-CD20 antibody is initiated.

[0020] In some embodiments of the method, the reference macrophage biomarker amount or level is the amount of macrophages measured by gene expression. In some embodiments of the method, the amount of macrophages is between approximately 0% and approximately 56.5%.

[0021] In some embodiments of the method, the benefit is an extension of progression-free survival (PFS). In some embodiments of the method, the benefit is an increase in overall survival (OS). In some embodiments of the method, the method may further include achieving an improvement in PFS or OS.

[0022] In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the macrophage biomarker is the average of the M1 macrophage gene signature set scores of one or more M1 macrophage gene signature sets. In some embodiments of the method, each M1 macrophage gene signature set score is the average of the expression levels of one or more genes in the M1 macrophage gene signature set. In some embodiments of the method, each M1 macrophage gene signature set score is the average of the normalized expression levels of one or more genes in the M1 macrophage gene signature set. In some embodiments of the method, one or more M1 macrophage gene signature sets include: (a) ACP2, ABCD1, C1QA, FDX1, CCL22, CD163, SCAMP2, ADAMDEC1, ARL8B, and HAMP; (b) ACP2, ABCD1, FDX1, CCL8, CCL22, CD163, ADAMDEC1, TREM2, and HAMP; (c) ACP2, ADRA2B , ALCAM, ABCD1, ATOX1, ATP6V0C, ATP6V1E1, BLVRA, C1QA, CD48, CD63, CLCN7, TPP1, CLTC, CCR1, CMKLR1, SLC31 A1, COX5B, FCER1G, FDX1, FOLR2, FPR3, FTL, HEXB, HK3, IL10, IL12B, ITGAE, LAIR1, CXCL9, MMP19, NARS, NDUFS 2, P2RX7, PDCL, MAPK13, PTGIR, PTPRA, RELA, CCL7, CCL8, CCL19, CCL22, SRC, STX4, TCEB1, TFRC, AGPS, MARCO , SNX3, CD84, USP14, ITGB1BP1, ATP6V1F, TRIP4, CD163, CIAO1, WTAP, ARHGEF11, ABI1, SCAMP2, ACTR2, BCAP31 , ZMPSTE24, BCKDK, EXOC5, STIP1, UQCR11, SDS, LILRB4, OGFR, TFEC, FKBP15, DNAJC13, TDRD7, STX12, IL17RA, ABTB2, FAM32A, SIGLEC7, SIGLEC9, ADAMDEC1, CECR5, SLC25A24, NRBP1, MS4A4A, TREM2, OTUD4, PQLC2, HAUS2,ARL8B, NECAP2, WDR11, ZC3H15, CCDC47, UTP3, MRS2, HAMP, MRPL40, VPS33A, CORO7, LIMD2, TMX1, DOT1L, ADO, and ADCK2, (d)ACP2, ADRA2B, ALCAM, TSPO, C3AR1, DAGLA, CALR, CHIT1, CYBB, CYC1, CYP19A1, DLAT, FCER1G, GP1BA, GPD1, IFNAR1, IL10, KCNJ5, KIFC3, MT2A, MYBPH, MYH11, MYO7A, P2RX7, PRDX1, RAB3IL1, RNH1, MRPL12, CCL1, CCL7, CCL8, CCL24, SRC, VIM, RRP1, MARCO, S1PR2, AP1M2, ACTR3, LILRB1, AFG3L2, SDS, LILRB4, EMILIN1, VSIG4, HSPB7, COQ2, ADAMDEC1, CECR5, WSB2, SLAMF8, DNASE2B, CLPB, MFSD7, and ADCK2, (e)ACP2, ADCY3, ADRA2B, ALCAM, TSPO, C1QA, C1QB, C3AR1, DAGLA, CD63, CHIT1, CMKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP19A1, FANCE, FCER1G, FDX1, FPR3, FTL, GP1BA, GPD1, HEXB, IL10, KCNJ1, KCNJ5, KIFC3, LAMP1, MMP19, MSR1, MT2A, MYBPH, MYO7A, P2RX7, PRDX1, RAB3IL1, MRPL12, CCL1, CCL7, CCL8, CCL18, CCL19, CCL24, SLC6A12, SPR, SRC, RRP1, MARCO, PKD2L1, S1PR2, CD163, LONP1, AP1M2, IGSF6, LILRB1, SDS, LILRB4, EMILIN1, VSIG4, TFEC, PHLDB1, CYFIP1, FKBP15, NCAPH, MYOF, HSPB7, ADAMDEC1, GLRX2, NDUFAF1, SPG21, MS4A4A, ATP6V1D, ATP6V1H, TREM2, PQLC2, TMEM70, PLEKHB2, TMEM33, SLAMF8, HAMP, DNASE2B, MYOZ1, LONRF3, CLPB, MFSD7, and ADCK2, and / or (f)ACP2, ADCY3, ADRA2B,ALCAM, ABCD1, ANXA2, ATP6V1A, C1QA, C1QB, C3AR1, DAGLA, CD80, CD63, CHIT1, CMKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP19A1, FANCE, FDX1, FPR 2, FPR3, GPD1, HEXB, KCNJ1, KCNJ5, KIFC3, MMP19, MSR1, MT2A, MYBPH, P2RX7, MAPK13, S100A11, CCL1, CCL7, CCL8, CCL18, CCL19, CCL22, CCL24, SLC1A2 These include SLC6A12, SLC11A1, SIGLEC1, SRC, TIE1, MARCO, HYAL2, CD163, LONP1, IGSF6, LILRB1, CD300C, SDS, LILRB4, EMILIN1, VSIG4, PHLDB1, NCAPH, CLEC4E, MYOF, HSPB7, ADAMDEC1, GLRX2, MS4A4A, ATP6V1H, TREM2, TMEM70, TMEM33, KCNK13, SLAMF8, HAMP, DNASE2B, MYOZ1, MFSD7, ADO, ADCK2, and TBC1D16.

[0023] In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the macrophage biomarker is the average TAM gene signature set score of one or more TAM gene signature sets. In some embodiments of the method, each TAM gene signature set score is the average expression level of one or more genes in the TAM gene signature set. In some embodiments of the method, each TAM gene signature set score is the average normalized expression level of one or more genes in the TAM gene signature set. In some embodiments of the method, one or more TAM gene signature sets are (a) MARCO, ACP5, VSIG4, MRC1, MSR1, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, C1QC, APOE, FOLR2, CTSD, and SPP1.

[0024] In some embodiments of the method, the macrophage biomarker is the gene expression level. In some embodiments of the method, the gene expression level is the median of the gene expression levels. In some embodiments of the method, the gene expression level is measured using a gene signature matrix. In some embodiments of the method, the gene signature matrix is ​​the following genes: (a) CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, A UTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FA M46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2 RX5, CEACAM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, HOPX, CD1D, GNG7, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD 11, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRC32, ARHGAP24, STAT4, SLC7A8 , CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PTGS1, TMEM170B, TREM2 , ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2,CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1; or (b) CD200, KLHL14, TCL1A, NRG1, CYP4F3, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, CD248, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, NINJ2, ABCB4, CD5, HAL, HPGD, BLNK, PLCL1, CEP19, HPSE, SLFN13, HOPX, CD1D, GNG7, TMEM154, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, TECPR2, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, NRGN, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRC32, ARHGAP24, PPP1R3B, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, CD36, ALDH1A2, ENPP2, COLEC12, PTGS1, TMEM170B, DOCK5, TREM2, C5AR2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1BIncludes MATK, LMO2, CFB, CCRL2, CLEC4A, TLR4, LILRA2, ACE, TLR1, LRRK2, LY96, NUPR1, CISH, CSTA, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1.

[0025] In some embodiments of the method, the gene signature matrix includes the following genes: CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VP REB3, DPEP3, MME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK 2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, H OPX, CD1D, GNG7, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, M GAM, TTC38, LRRC32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLE It consists of C12, PTGS1, TMEM170B, TREM2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1.In some embodiments of the method, a gene signature matrix is ​​used to determine the number of M1 macrophages or tumor-associated macrophages.

[0026] In some embodiments of the methods, which may be combined with any of the aforementioned aspects or embodiments, the macrophage biomarker is a certain amount of M1 macrophages or a certain amount of tumor-associated macrophages. In some embodiments of the methods, the amount of M1 macrophages or tumor-associated macrophages is measured directly or indirectly. In some embodiments of the methods, the amount of M1 macrophages or tumor-associated macrophages is measured directly using flow cytometry, spatial transcriptomics, spatial proteomics, or a combination thereof. In some embodiments of the methods, the amount of M1 macrophages or tumor-associated macrophages is measured indirectly using nucleic acids or proteins. In some embodiments of the methods, nucleic acids are measured using RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof. In some embodiments of the methods, the amount of M1 macrophages or tumor-associated macrophages is measured using a marker gene approach or a deconvolution approach. In some embodiments of the methods, the marker gene approach uses xCell. In some embodiments of the method, the deconvolution approach uses quanTIseq.

[0027] In some embodiments of the method, macrophage biomarkers in patient samples are measured using nucleic acids or proteins. In some embodiments of the method, macrophage biomarkers in patient samples are determined using nucleic acid expression levels. In some embodiments of the method, nucleic acid expression levels are determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof. In some embodiments of the method, nucleic acid expression levels are mRNA expression levels. In some embodiments of the method, mRNA expression levels are determined by RNA-seq.

[0028] In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof. In some embodiments of the method, the sample is a tissue sample. In some embodiments of the method, the tissue sample is a tumor tissue sample. In some embodiments of the method, the tumor tissue sample includes tumor cells, tumor-infiltrating immune cells, stromal cells, normal adjacent tissue (NAT) cells, or a combination thereof. In some embodiments of the method, the tumor tissue sample is a biopsy sample. In some embodiments of the method, the sample is an archived sample, a fresh sample, or a frozen sample.

[0029] In some embodiments of the methods that can be combined with any of the aforementioned embodiments or embodiments, DLBCL is a subgroup of DLBCL originating from germinal center B-cell-like (GCB) or activated B-cell-like (ABC) cells. In some embodiments of the methods that can be combined with any of the aforementioned embodiments or embodiments, DLBCL is CD79b-positive and / or CD20-positive DLBCL. In some embodiments of the methods that can be combined with any of the aforementioned embodiments or embodiments, the patient has not been previously treated for DLBCL. In some embodiments of the methods that can be combined with any of the aforementioned embodiments or embodiments, the patient has not been previously administered immunoconjugates and anti-CD20 antibodies.

[0030] In some embodiments of the method that can be combined with any of the aforementioned embodiments or embodiments, the anti-CD79b antibody comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) containing the amino acid sequence of SEQ ID NO: 4. In some embodiments of the method that can be combined with any of the aforementioned embodiments or embodiments, the anti-CD79b antibody comprises (a) a heavy chain containing the amino acid sequence of SEQ ID NO: 13 and a light chain containing the amino acid sequence of SEQ ID NO: 11, (b) a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 14, or (c) a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11. In some embodiments of the method that can be combined with any of the aforementioned embodiments or embodiments, p is 2-7, 2-6, 2-5, 3-5, or 3-4. In some embodiments of the method, p is 3.4. In some embodiments of the method, p is 3.5. In some embodiments of the method that can be combined with any of the aforementioned embodiments or embodiments, the immunoconjugate is polatuzumab vedotin.

[0031] In some embodiments of the method, which can be combined with any of the aforementioned embodiments or models, the anti-CD20 antibody is a type I anti-CD20 antibody or a type II anti-CD20 antibody. In some embodiments of the method, the anti-CD20 antibody is a type I anti-CD20 antibody. In some embodiments of the method, the type I anti-CD20 antibody includes the following CDRs: (a) CDR-H1 having the amino acid sequence of SEQ ID NO: 26, (b) CDR-H2 having the amino acid sequence of SEQ ID NO: 27, (c) CDR-H3 having the amino acid sequence of SEQ ID NO: 28, (d) CDR-L1 having the amino acid sequence of SEQ ID NO: 29, (e) CDR-L2 having the amino acid sequence of SEQ ID NO: 30, and (f) CDR-L3 having the amino acid sequence of SEQ ID NO: 31. In some embodiments of the method, the type I anti-CD20 antibody includes a VH domain containing the amino acid sequence of SEQ ID NO: 40 and a VL domain containing the amino acid sequence of SEQ ID NO: 41. In some embodiments of the method, the type I anti-CD20 antibody is rituximab.

[0032] In some embodiments of the method, polatuzumab vedotin is administered at a dose of approximately 1.0 mg / kg to approximately 1.8 mg / kg. In some embodiments of the method, polatuzumab vedotin is administered at a dose of approximately 1.8 mg / kg. In some embodiments of the method, rituximab is administered at approximately 375 mg / m². 2 It is administered in the following dose. In some embodiments, polatuzumab vedotin and / or rituximab are administered intravenously.

[0033] In some embodiments of the method, the method may further include administering an effective amount of an additional therapeutic agent to the patient. In some embodiments of the method, the additional therapeutic agent is one or more of the following: chemotherapeutic agents, corticosteroids, antineoplastic agents, growth inhibitors, anti-angiogenic agents, radiotherapy, cytotoxic agents, or combinations thereof. In some embodiments of the method, the additional therapeutic agent is a chemotherapeutic agent and a corticosteroid. In some embodiments of the method, the chemotherapeutic agent is cyclophosphamide and / or doxorubicin. In some embodiments of the method, the corticosteroid is prednisone, prednisolone, or methylprednisolone. In some embodiments of the method, the cyclophosphamide is approximately 375 mg / m² 2 ~about 750mg / m 2 It is administered in the following dose. In some embodiments, doxorubicin is administered at approximately 25 mg / m². 2 ~about 50mg / m 2 The following doses are administered: In some embodiments, (a) prednisone is administered at a dose of about 100 mg, or (b) prednisolone is administered at a dose of about 100 mg, or (c) methylprednisolone is administered at a dose of about 80 mg. In some embodiments, cyclophosphamide and / or doxorubicin are administered intravenously.

[0034] In some embodiments of the method, prednisone, prednisolone, or methylprednisolone is administered orally. In some embodiments of the method, polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone, prednisolone, or methylprednisolone is administered in at least one 21-day cycle. In some embodiments, (a) polatuzumab vedotin, rituximab, cyclophosphamide, and / or doxorubicin is administered on day 1 of each 21-day cycle, and / or (b) prednisone, prednisolone, or methylprednisolone is administered on days 1 to 5 of each 21-day cycle. In some embodiments of the method, polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone, prednisolone, or methylprednisolone are administered in 1, 2, 3, 4, 5, or 6 21-day cycles.

[0035] In some embodiments, the present invention relates to the use of an immunoconjugate and an anti-CD20 antibody in the manufacture of a pharmaceutical for the treatment of DLBCL, for treating patients in whom the amount or level of a macrophage biomarker in a sample from a patient is below the amount or level of a reference macrophage biomarker, wherein the immunoconjugate has the following formula: [ka] (wherein Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8) is characterized by use.

[0036] In some embodiments of use, the patient is a human patient.

[0037] In some embodiments of use, which may be combined with any of the embodiments or models described above, the reference macrophage biomarker quantity or level is a pre-assigned macrophage biomarker quantity or level. In some embodiments of use, which may be combined with any of the embodiments or models described above, the reference macrophage biomarker quantity or level is the quantity or level of macrophage biomarkers in a reference population. In some embodiments of use, the quantity or level of macrophage biomarkers in a reference population is the median quantity or level of macrophage biomarkers in the reference population.

[0038] In some embodiments of use, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 25th percentile of the reference population. In some embodiments of use, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 50th percentile of the reference population. In some embodiments of use, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 75th percentile of the reference population.

[0039] In some embodiments of use, the reference population is a population of patients with DLBCL. In some embodiments of use, the population of patients with DLBCL has been previously treated with an immunoconjugate and an anti-CD20 antibody. In some embodiments of use, the population of patients with DLBCL has been previously treated with an anti-CD20 antibody. In some embodiments of use, the reference macrophage biomarker amount or level is the amount or level of macrophage biomarkers in the reference population before initiating treatment with the immunoconjugate and anti-CD20 antibody.

[0040] In some embodiments of use, which may be combined with any of the aforementioned aspects or embodiments, the reference macrophage biomarker amount or level is the amount of macrophages measured by gene expression. In some embodiments of use, the amount of macrophages is between approximately 0% and approximately 56.5%.

[0041] In some embodiments of use that can be combined with any of the aforementioned aspects or embodiments, the treatment achieves an improvement in PFS or OS.

[0042] In some embodiments of use, which may be combined with any of the aforementioned aspects or embodiments, the macrophage biomarker is the mean of the M1 macrophage gene signature set scores of one or more M1 macrophage gene signature sets. In some embodiments of use, each M1 macrophage gene signature set score is the mean of the expression levels of one or more genes in the M1 macrophage gene signature set. In some embodiments of use, each M1 macrophage gene signature set score is the mean of the normalized expression levels of one or more genes in the M1 macrophage gene signature set. In some embodiments of use, one or more M1 macrophage gene signature sets include: (a) ACP2, ABCD1, C1QA, FDX1, CCL22, CD163, SCAMP2, ADAMDEC1, ARL8B, and HAMP; (b) ACP2, ABCD1, FDX1, CCL8, CCL22, CD163, ADAMDEC1, TREM2, and HAMP; (c) ACP2, ADRA2B , ALCAM, ABCD1, ATOX1, ATP6V0C, ATP6V1E1, BLVRA, C1QA, CD48, CD63, CLCN7, TPP1, CLTC, CCR1, CMKLR1, SLC31 A1, COX5B, FCER1G, FDX1, FOLR2, FPR3, FTL, HEXB, HK3, IL10, IL12B, ITGAE, LAIR1, CXCL9, MMP19, NARS, NDUFS 2, P2RX7, PDCL, MAPK13, PTGIR, PTPRA, RELA, CCL7, CCL8, CCL19, CCL22, SRC, STX4, TCEB1, TFRC, AGPS, MARCO , SNX3, CD84, USP14, ITGB1BP1, ATP6V1F, TRIP4, CD163, CIAO1, WTAP, ARHGEF11, ABI1, SCAMP2, ACTR2, BCAP31 , ZMPSTE24, BCKDK, EXOC5, STIP1, UQCR11, SDS, LILRB4, OGFR, TFEC, FKBP15, DNAJC13, TDRD7, STX12, IL17RA, ABTB2, FAM32A, SIGLEC7, SIGLEC9, ADAMDEC1, CECR5, SLC25A24, NRBP1, MS4A4A, TREM2, OTUD4, PQLC2, HAUS2,ARL8B, NECAP2, WDR11, ZC3H15, CCDC47, UTP3, MRS2, HAMP, MRPL40, VPS33A, CORO7, LIMD2, TMX1, DOT1L, ADO, and ADCK2, (d)ACP2, ADRA2B, ALCAM, TSPO, C3AR1, DAGLA, CALR, CHIT1, CYBB, CYC1, CYP19A1, DLAT, FCER1G, GP1BA, GPD1, IFNAR1, IL10, KCNJ5, KIFC3, MT2A, MYBPH, MYH11, MYO7A, P2RX7, PRDX1, RAB3IL1, RNH1, MRPL12, CCL1, CCL7, CCL8, CCL24, SRC, VIM, RRP1, MARCO, S1PR2, AP1M2, ACTR3, LILRB1, AFG3L2, SDS, LILRB4, EMILIN1, VSIG4, HSPB7, COQ2, ADAMDEC1, CECR5, WSB2, SLAMF8, DNASE2B, CLPB, MFSD7, and ADCK2, (e)ACP2, ADCY3, ADRA2B, ALCAM, TSPO, C1QA, C1QB, C3AR1, DAGLA, CD63, CHIT1, CMKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP19A1, FANCE, FCER1G, FDX1, FPR3, FTL, GP1BA, GPD1, HEXB, IL10, KCNJ1, KCNJ5, KIFC3, LAMP1, MMP19, MSR1, MT2A, MYBPH, MYO7A, P2RX7, PRDX1, RAB3IL1, MRPL12, CCL1, CCL7, CCL8, CCL18, CCL19, CCL24, SLC6A12, SPR, SRC, RRP1, MARCO, PKD2L1, S1PR2, CD163, LONP1, AP1M2, IGSF6, LILRB1, SDS, LILRB4, EMILIN1, VSIG4, TFEC, PHLDB1, CYFIP1, FKBP15, NCAPH, MYOF, HSPB7, ADAMDEC1, GLRX2, NDUFAF1, SPG21, MS4A4A, ATP6V1D, ATP6V1H, TREM2, PQLC2, TMEM70, PLEKHB2, TMEM33, SLAMF8, HAMP, DNASE2B, MYOZ1, LONRF3, CLPB, MFSD7, and ADCK2, and / or (f)ACP2, ADCY3, ADRA2B,ALCAM, ABCD1, ANXA2, ATP6V1A, C1QA, C1QB, C3AR1, DAGLA, CD80, CD63, CHIT1, CMKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP19A1, FANCE, FDX1, FPR 2, FPR3, GPD1, HEXB, KCNJ1, KCNJ5, KIFC3, MMP19, MSR1, MT2A, MYBPH, P2RX7, MAPK13, S100A11, CCL1, CCL7, CCL8, CCL18, CCL19, CCL22, CCL24, SLC1A2 These include SLC6A12, SLC11A1, SIGLEC1, SRC, TIE1, MARCO, HYAL2, CD163, LONP1, IGSF6, LILRB1, CD300C, SDS, LILRB4, EMILIN1, VSIG4, PHLDB1, NCAPH, CLEC4E, MYOF, HSPB7, ADAMDEC1, GLRX2, MS4A4A, ATP6V1H, TREM2, TMEM70, TMEM33, KCNK13, SLAMF8, HAMP, DNASE2B, MYOZ1, MFSD7, ADO, ADCK2, and TBC1D16.

[0043] In some embodiments of use, which may be combined with any of the embodiments described above, the macrophage biomarker is the mean of the TAM gene signature set score of one or more tumor-associated macrophage (TAM) gene signature sets. In some embodiments of use, each TAM gene signature set score is the mean of the expression levels of one or more genes in the TAM gene signature set. In some embodiments of use, each TAM gene signature set score is the mean of the normalized expression levels of one or more genes in the TAM gene signature set. In some embodiments of use, one or more TAM gene signature sets are MARCO, ACP5, VSIG4, MRC1, MSR1, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, C1QC, APOE, FOLR2, CTSD, and SPP1.

[0044] In some embodiments of use, which may be combined with any of the aforementioned aspects or embodiments, the macrophage biomarker is a gene expression value. In some embodiments of use, the gene expression value is the median of the gene expression values. In some embodiments of use, the gene expression value is measured using a gene signature matrix. In some embodiments of use, the gene signature matrix includes the following genes: (a) CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WN T5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, T XK, CD8B, P2RX5, CEACAM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, HOPX, CD1D, GNG7, TCF4, BANK1, FHIT, FCMR, GNG 2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRC32, ARHGAP 24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PT GS1, TMEM170B, TREM2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B,MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1, or (b) CD200, KLHL14, TCL1A, NRG1, CYP4F3, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, CD248, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, NINJ2, ABCB4, CD5, HAL, HPGD, BLNK, PLCL1, CEP19, HPSE, SLFN13, HOPX, CD1D, GNG7, TMEM154, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, TECPR2, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, NRGN, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRC32, ARHGAP24, PPP1R3B, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, CD36, ALDH1A2, ENPP2, COLEC12, PTGS1, TMEM170B, DOCK5, TREM2, C5AR2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2,Includes ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, TLR4, LILRA2, ACE, TLR1, LRRK2, LY96, NUPR1, CISH, CSTA, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1.

[0045] In some embodiments of use, the gene signature matrix includes the following genes: CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VP REB3, DPEP3, MME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK 2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, H OPX, CD1D, GNG7, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, M GAM, TTC38, LRRC32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLE It consists of C12, PTGS1, TMEM170B, TREM2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1.In some embodiments of use, the gene signature matrix is ​​used to determine the number of M1 macrophages or tumor-associated macrophages.

[0046] In some embodiments of use, which may be combined with any of the embodiments described above, the macrophage biomarker is a certain amount of M1 macrophages or a certain amount of tumor-associated macrophages. In some embodiments of use, the amount of M1 macrophages or tumor-associated macrophages is measured directly or indirectly. In some embodiments of use, the amount of M1 macrophages or tumor-associated macrophages is measured directly using flow cytometry, spatial transcriptomics, spatial proteomics, or a combination thereof. In some embodiments of use, the amount of M1 macrophages or tumor-associated macrophages is measured indirectly using nucleic acids or proteins. In some embodiments of use, nucleic acids are measured using RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof. In some embodiments of use, the amount of M1 macrophages or tumor-associated macrophages is measured using a marker gene approach or a deconvolution approach. In some embodiments of use, the marker gene approach uses xCell. In some embodiments of use, the deconvolution approach uses quanTIseq.

[0047] In some embodiments of use, macrophage biomarkers in patient samples are measured using nucleic acids or proteins. In some embodiments of use, macrophage biomarkers in patient samples are determined using nucleic acid expression levels. In some embodiments of use, nucleic acid expression levels are determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof. In some embodiments of use, nucleic acid expression levels are mRNA expression levels. In some embodiments of use, mRNA expression levels are determined by RNA-seq.

[0048] In some embodiments of use, which may be combined with any of the aforementioned aspects or embodiments, the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof. In some embodiments of use, the sample is a tissue sample. In some embodiments of use, the tissue sample is a tumor tissue sample. In some embodiments of use, the tumor tissue sample includes tumor cells, tumor-infiltrating immune cells, stromal cells, normal adjacent tissue (NAT) cells, or a combination thereof. In some embodiments of use, the tumor tissue sample is a biopsy sample. In some embodiments of use, the sample is an archived sample, a fresh sample, or a frozen sample.

[0049] In some embodiments of use that can be combined with any of the aforementioned embodiments or embodiments, DLBCL is a subgroup of DLBCL originating from germinal center B-cell-like (GCB) or activated B-cell-like (ABC) cells. In some embodiments of use that can be combined with any of the aforementioned embodiments or embodiments, DLBCL is CD79a-positive and / or CD20-positive DLBCL. In some embodiments of use that can be combined with any of the aforementioned embodiments or embodiments, the patient has not been previously treated for DLBCL. In some embodiments of use that can be combined with any of the aforementioned embodiments or embodiments, the patient has not been previously administered immunoconjugates and anti-CD20 antibodies.

[0050] In some embodiments of use that can be combined with any of the aforementioned aspects or embodiments, the anti-CD79b antibody comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) containing the amino acid sequence of SEQ ID NO: 4. In some embodiments of use, the anti-CD79b antibody comprises (a) a heavy chain containing the amino acid sequence of SEQ ID NO: 13 and a light chain containing the amino acid sequence of SEQ ID NO: 11, (b) a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 14, or (c) a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11. In some embodiments of use that can be combined with any of the aforementioned aspects or embodiments, p is 2-7, 2-6, 2-5, 3-5, or 3-4. In some embodiments of use, p is 3.4. In some embodiments of use, p is 3.5. In some embodiments of use that can be combined with any of the aforementioned aspects or embodiments, the immunoconjugate is polatuzumab vedotin.

[0051] In some embodiments of use, which may be combined with any of the aforementioned embodiments or models, the anti-CD20 antibody is a type I anti-CD20 antibody or a type II anti-CD20 antibody. In some embodiments of use, the anti-CD20 antibody is a type I anti-CD20 antibody. In some embodiments of use, the type I anti-CD20 antibody includes the following CDRs: (a) CDR-H1 having the amino acid sequence of SEQ ID NO: 26, (b) CDR-H2 having the amino acid sequence of SEQ ID NO: 27, (c) CDR-H3 having the amino acid sequence of SEQ ID NO: 28, (d) CDR-L1 having the amino acid sequence of SEQ ID NO: 29, (e) CDR-L2 having the amino acid sequence of SEQ ID NO: 30, and (f) CDR-L3 having the amino acid sequence of SEQ ID NO: 31. In some embodiments of use, the type I anti-CD20 antibody includes a VH domain containing the amino acid sequence of SEQ ID NO: 40 and a VL domain containing the amino acid sequence of SEQ ID NO: 41. In some embodiments of use, the type I anti-CD20 antibody is rituximab.

[0052] In some embodiments of use, polatuzumab vedotin is administered at a dose of about 1.0 mg / kg to about 1.8 mg / kg. In some embodiments of use, polatuzumab vedotin is administered at a dose of about 1.8 mg / kg. In some embodiments of use, rituximab is administered at a dose of about 375 mg / m 2 . In some embodiments of use, polatuzumab vedotin and / or rituximab are administered intravenously.

[0053] In some embodiments of use that can be combined with any of the foregoing aspects or embodiments, the medicament is administered to a patient in combination with an effective amount of an additional therapeutic agent. In some embodiments of use, the additional therapeutic agent is one or more of a chemotherapeutic agent, a corticosteroid, an anti-neoplastic agent, a growth inhibitor, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some embodiments of use, the additional therapeutic agent is a chemotherapeutic agent and a corticosteroid. In some embodiments of use, the chemotherapeutic agent is cyclophosphamide and / or doxorubicin. In some embodiments of use, the corticosteroid is prednisone, prednisolone or methylprednisolone. In some embodiments of use, cyclophosphamide is administered at a dose of about 375 mg / m 2 to about 750 mg / m 2 . In some embodiments of use, doxorubicin is administered at a dose of about 25 mg / m 2 to about 50 mg / m 2 . In some embodiments of use, (a) prednisone is administered at a dose of about 100 mg, or (b) prednisolone is administered at a dose of about 100 mg, or (c) methylprednisolone is administered at a dose of about 80 mg. In some embodiments of use, cyclophosphamide and / or doxorubicin are administered intravenously. In some embodiments of use, prednisone, prednisolone or methylprednisolone are administered orally.

[0054] In some embodiments of use, polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone, prednisolone, or methylprednisolone are administered in at least one 21-day cycle. In some embodiments of use, (a) polatuzumab vedotin, rituximab, cyclophosphamide, and / or doxorubicin are administered on day 1 of each 21-day cycle, and / or (b) prednisone, prednisolone, or methylprednisolone are administered on days 1 to 5 of each 21-day cycle. In some embodiments of use, polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone, prednisolone, or methylprednisolone are administered in one, two, three, four, five, or six 21-day cycles.

[0055] In some embodiments, an immunoconjugate and an anti-CD20 antibody for use in the treatment of patients having DLBCL and in which the amount or level of macrophage biomarkers in a patient sample is below the reference amount or level of macrophage biomarkers, wherein the immunoconjugate has the following formula: [ka] The present invention is characterized by an immunoconjugate containing (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8) and an anti-CD20 antibody.

[0056] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the patient is a human patient.

[0057] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the reference macrophage biomarker amount or level is a pre-assigned macrophage biomarker amount or level. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the reference macrophage biomarker amount or level is the amount or level of macrophage biomarkers in a reference population. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the amount or level of macrophage biomarkers in a reference population is the median amount or level of macrophage biomarkers in a reference population.

[0058] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 25th percentile of the reference population. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 50th percentile of the reference population. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 75th percentile of the reference population.

[0059] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the reference population is a population of patients with DLBCL. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the population of patients with DLBCL has been previously treated with the immunoconjugates and anti-CD20 antibodies. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the population of patients with DLBCL has been previously treated with the anti-CD20 antibodies. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the reference macrophage biomarker amount or level is the amount or level of macrophage biomarkers in the reference population before initiating treatment with the immunoconjugates and anti-CD20 antibodies.

[0060] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned aspects or embodiments, the reference macrophage biomarker amount or level is the amount of macrophages measured by gene expression. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the amount of macrophages is between approximately 0% and approximately 56.5%.

[0061] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned aspects or embodiments, the treatment achieves improvement in PFS or OS.

[0062] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the macrophage biomarker is the average of the M1 macrophage gene signature set scores of one or more M1 macrophage gene signature sets. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, each M1 macrophage gene signature set score is the average of the expression levels of one or more genes in the M1 macrophage gene signature set. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, each M1 macrophage gene signature set score is the average of the normalized expression levels of one or more genes in the M1 macrophage gene signature set. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, one or more M1 macrophage gene signature sets include: (a) ACP2, ABCD1, C1QA, FDX1, CCL22, CD163, SCAMP2, ADAMDEC1, ARL8B, and HAMP; (b) ACP2, ABCD1, FDX1, CCL8, CCL22, CD163, ADAMDEC1, TREM2, and HAMP; (c) ACP2, ADRA2B, ALCAM, ABCD1, ATOX1, ATP6V0C, ATP6V1E1, BLVRA, C1QA, CD48, CD63, CLCN7, TPP1, CLTC, CCR1, CMKLR1, SLC31A1, COX5B, FCER1G, FDX1, FOLR2, FPR3, FT L, HEXB, HK3, IL10, IL12B, ITGAE, LAIR1, CXCL9, MMP19, NARS, NDUFS2, P2RX7, PDCL, MAPK13, PTGIR , PTPRA, RELA, CCL7, CCL8, CCL19, CCL22, SRC, STX4, TCEB1, TFRC, AGPS, MARCO, SNX3, CD84, USP14, ITGB1BP1, ATP6V1F, TRIP4, CD163, CIAO1, WTAP, ARHGEF11, ABI1, SCAMP2, ACTR2, BCAP31, ZMPSTE2 4, BCKDK, EXOC5, STIP1, UQCR11, SDS, LILRB4, OGFR, TFEC, FKBP15, DNAJC13, TDRD7, STX12, IL17RA,ABTB2、FAM32A、SIGLEC7、SIGLEC9、ADAMDEC1、CECR5、SLC25A24、NRBP1、MS4 A4A、TREM2、OTUD4、PQLC2、HAUS2、ARL8B、NECAP2、WDR11、ZC3H15、CCDC47、U TP3, MRS2, HAMP, MRPL40, VPS33A, CORO7, LIMD2, TMX1, DOT1L, ADO, ADCK2, (d)ACP2, ADRA2B, ALCAM, TSPO, C3AR1, DAGLA, CALR, CHIT1, CYBB, CYC1, CYP 19A1、DLAT、FCER1G、GP1BA、GPD1、IFNAR1、IL10、KCNJ5、KIFC3、MT2A、MYBPH 、MYH11、MYO7A、P2RX7、PRDX1、RAB3IL1、RNH1、MRPL12、CCL1、CCL7、CCL8、CC L24、SRC、VIM、RRP1、MARCO、S1PR2、AP1M2、ACTR3、LILRB1、AFG3L2、SDS、LIL RB4、EMILIN1、VSIG4、HSPB7、COQ2、ADAMDEC1、CECR5、WSB2、SLAMF8、DNASE2B CLPB, MFSD7, ADCK2, (e)ACP2, ADCY3, ADRA2B, ALCAM, TSPO, C1QA, C1QB, C3AR1, DAGLA, CD63, CHIT1, CMLKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP 19A1、FANCE、FCER1G、FDX1、FPR3、FTL、GP1BA、GPD1、HEXB、IL10、KCNJ1、KCN J5、KIFC3、LAMP1、MMP19、MSR1、MT2A、MYBPH、MYO7A、P2RX7、PRDX1、RAB3IL1、 MRPL12、CCL1、CCL7、CCL8、CCL18、CCL19、CCL24、SLC6A12、SPR、SRC、RRP1、M ARCO、PKD2L1、S1PR2、CD163、LONP1、AP1M2、IGSF6、LILRB1、SDS、LILRB4、EMI LIN1、VSIG4、TFEC、PHLDB1、CYFIP1、FKBP15、NCAPH、MYOF、HSPB7、ADAMDEC1 、GLRX2、NDUFAF1、SPG21、MS4A4A、ATP6V1D、ATP6V1H、TREM2、PQLC2、TMEM70、PLEKHB2, TMEM33, SLAMF8, HAMP, DNASE2B, MYOZ1, LONRF3, CLPB, MFSD7, and ADCK2, and / or (f)ACP2, ADCY3, ADRA2B, ALCAM, ABCD1, ANXA2, ATP6V1A, C1QA, C1QB, C3AR1, DAGLA, CD80, CD63, CHIT1, CMKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP19A1, FANCE, FDX1, FPR2, FPR3, GPD1, HEXB, KCNJ1, KCNJ5, KIFC3, MMP19, MSR1, MT2A, MYBPH, P2RX7, MAPK13, S100A1 These are CCL1, CCL7, CCL8, CCL18, CCL19, CCL22, CCL24, SLC1A2, SLC6A12, SLC11A1, SIGLEC1, SRC, TIE1, MARCO, HYAL2, CD163, LONP1, IGSF6, LILRB1, CD300C, SDS, LILRB4, EMILIN1, VSIG4, PHLDB1, NCAPH, CLEC4E, MYOF, HSPB7, ADAMDEC1, GLRX2, MS4A4A, ATP6V1H, TREM2, TMEM70, TMEM33, KCNK13, SLAMF8, HAMP, DNASE2B, MYOZ1, MFSD7, ADO, ADCK2, and TBC1D16.

[0063] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the macrophage biomarker is the average of the tumor-associated macrophage gene signature set scores of one or more tumor-associated macrophage gene signature sets. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, each tumor-associated macrophage gene signature set score is the average of the expression levels of one or more genes in the tumor-associated macrophage gene signature set. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, each tumor-associated macrophage gene signature set score is the average of the normalized expression levels of one or more genes in the tumor-associated macrophage gene signature set. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, one or more tumor-associated macrophage gene signature sets are (a) MARCO, ACP5, VSIG4, MRC1, MSR1, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, C1QC, APOE, FOLR2, CTSD, and SPP1.

[0064] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the macrophage biomarker is the gene expression value. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the gene expression value is the median of the gene expression values. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the gene expression value is measured using a gene signature matrix. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the gene signature matrix is ​​the following genes: (a) CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A 3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SE MA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLCL1 , HPSE, SLFN13, HOPX, CD1D, GNG7, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAO A, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRC32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SM ARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PTGS1, TMEM170B, TREM2, ECM1, SLC1A3, ABHD5,MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1, or (b) CD200, KLHL14, TCL1A, NRG1, CYP4F3, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, CD248, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, NINJ2, ABCB4, CD5, HAL, HPGD, BLNK, PLCL1, CEP19, HPSE, SLFN13, HOPX, CD1D, GNG7, TMEM154, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, TECPR2, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, NRGN, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRC32, ARHGAP24, PPP1R3B, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, CD36, ALDH1A2, ENPP2, COLEC12, PTGS1, TMEM170B, DOCK5, TREM2, C5AR2,It includes ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, TLR4, LILRA2, ACE, TLR1, LRRK2, LY96, NUPR1, CISH, CSTA, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1.,

[0065] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the gene signature matrix includes the following genes: CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A , ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PL CL1, HPSE, SLFN13, HOPX, CD1D, GNG7, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, DGA T2, FXYD6, TMCC3, MGAM, TTC38, LRRC32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALD H1A2, ENPP2, COLEC12, PTGS1, TMEM170B, TREM2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KD M1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA,It consists of and ICAM1. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, a gene signature matrix is ​​used to determine the number of M1 macrophages or tumor-associated macrophages.

[0066] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned aspects or embodiments, the macrophage biomarker is the amount of M1 macrophages or tumor-associated macrophages. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the amount of M1 macrophages or tumor-associated macrophages is measured directly or indirectly. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the amount of M1 macrophages or tumor-associated macrophages is measured directly using flow cytometry, spatial transcriptomics, spatial proteomics, or a combination thereof. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the amount of M1 macrophages or tumor-associated macrophages is measured indirectly using nucleic acids or proteins. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, nucleic acids are measured using RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the amount of M1 macrophages or tumor-associated macrophages is measured using a marker gene approach or a deconvolution approach. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the marker gene approach uses xCell. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the deconvolution approach uses quanTIseq.

[0067] In some embodiments of immunoconjugates and anti-CD20 antibodies for use, macrophage biomarkers in patient samples are measured using nucleic acids or proteins. In some embodiments of immunoconjugates and anti-CD20 antibodies for use, macrophage biomarkers in patient samples are determined using nucleic acid expression levels. In some embodiments of immunoconjugates and anti-CD20 antibodies for use, nucleic acid expression levels are determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof. In some embodiments of immunoconjugates and anti-CD20 antibodies for use, nucleic acid expression levels are mRNA expression levels. In some embodiments of immunoconjugates and anti-CD20 antibodies for use, mRNA expression levels are determined by RNA-seq.

[0068] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the sample is a tissue sample. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the tissue sample is a tumor tissue sample. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the tumor tissue sample contains tumor cells, tumor-infiltrating immune cells, stromal cells, normal adjacent tissue (NAT) cells, or a combination thereof. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the tumor tissue sample is a biopsy sample. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the sample is an archived sample, a fresh sample, or a frozen sample.

[0069] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the sample is a tissue sample, and DLBCL is a subgroup of DLBCL originating from germinal center B-cell-like (GCB) or activated B-cell-like (ABC) cells. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, DLBCL is CD79a-positive and / or CD20-positive DLBCL. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the patient has not been previously treated for DLBCL. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the patient has not been previously administered the immunoconjugate and anti-CD20 antibody.

[0070] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which can be combined with any of the aforementioned embodiments or models, the sample is a tissue sample, and the anti-CD79b antibody comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) containing the amino acid sequence of SEQ ID NO: 4. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the anti-CD79b antibody comprises (a) a heavy chain containing the amino acid sequence of SEQ ID NO: 13 and a light chain containing the amino acid sequence of SEQ ID NO: 11, (b) a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 14, or (c) a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which can be combined with any of the aforementioned embodiments or models, p is 2-7, 2-6, 2-5, 3-5, or 3-4. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, p is 3.4. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, p is 3.5. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the above-described embodiments or models, the immunoconjugate is polatuzumab vedotin.

[0071] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use that can be combined with any of the aforementioned embodiments or models, the sample is a tissue sample and the anti-CD20 antibody is a type I anti-CD20 antibody or a type II anti-CD20 antibody. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the anti-CD20 antibody is a type I anti-CD20 antibody. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the type I anti-CD20 antibody includes the following CDRs: (a) CDR-H1 having the amino acid sequence of SEQ ID NO: 26, (b) CDR-H2 having the amino acid sequence of SEQ ID NO: 27, (c) CDR-H3 having the amino acid sequence of SEQ ID NO: 28, (d) CDR-L1 having the amino acid sequence of SEQ ID NO: 29, (e) CDR-L2 having the amino acid sequence of SEQ ID NO: 30, and (f) CDR-L3 having the amino acid sequence of SEQ ID NO: 31. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the type I anti-CD20 antibody comprises a VH domain containing the amino acid sequence of SEQ ID NO: 40 and a VL domain containing the amino acid sequence of SEQ ID NO: 41. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the type I anti-CD20 antibody is rituximab.

[0072] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, polatuzumab vedotin is intended for use at a dose of approximately 1.0 mg / kg to approximately 1.8 mg / kg. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, polatuzumab vedotin is intended for use at a dose of approximately 1.8 mg / kg. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, rituximab is intended for use at approximately 375 mg / m². 2 It is intended for use in the specified dose. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, polatuzumab vedotin and / or rituximab are intended for intravenous use.

[0073] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, which may be combined with any of the aforementioned embodiments or models, the treatment further includes the use of an effective amount of an additional therapeutic agent. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the additional therapeutic agent is one or more of the following: chemotherapeutic agents, corticosteroids, antineoplastic agents, growth inhibitors, anti-angiogenic agents, radiotherapy, cytotoxic agents, or a combination thereof. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the additional therapeutic agent is a chemotherapeutic agent and a corticosteroid. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the chemotherapeutic agent is cyclophosphamide and / or doxorubicin. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the corticosteroid is prednisone, prednisolone, or methylprednisolone. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, the amount of cyclophosphamide is about 375 mg / m². 2 ~about 750mg / m 2 This is intended for use in doses of approximately 25 mg / m². In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, doxorubicin is used in doses of approximately 25 mg / m². 2 ~about 50mg / m 2 These are intended for use in doses of approximately 100 mg. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, (a) prednisone is intended for use in doses of approximately 100 mg, or (b) prednisolone is intended for use in doses of approximately 100 mg, or (c) methylprednisolone is intended for use in doses of approximately 80 mg. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, cyclophosphamide and / or doxorubicin are intended for intravenous use. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, prednisone, prednisolone, or methylprednisolone are intended for oral use.

[0074] In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin and / or prednisone, prednisolone or methylprednisolone are intended for use in at least one 21-day cycle. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, (a) polatuzumab vedotin, rituximab, cyclophosphamide and / or doxorubicin are intended for use on day 1 of each 21-day cycle, and / or (b) prednisone, prednisolone or methylprednisolone are intended for use on days 1 to 5 of each 21-day cycle. In some embodiments relating to immunoconjugates and anti-CD20 antibodies for use, polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone, prednisolone, or methylprednisolone are intended for use in one, two, three, four, five, or six 21-day cycles.

[0075] In some embodiments, the present invention provides a method for identifying, diagnosing and / or predicting whether a patient with diffuse large B-cell lymphoma (DLBCL) can benefit from a treatment comprising an immunoconjugate, an anti-CD20 antibody, a chemotherapeutic agent and a corticosteroid, comprising measuring a macrophage biomarker in a sample from the patient, thereby identifying, diagnosing and / or predicting the patient to benefit from a treatment comprising an immunoconjugate, an anti-CD20 antibody, a chemotherapeutic agent and a corticosteroid, if the amount or level of the macrophage biomarker in the sample is below a reference macrophage biomarker amount or level, and the immunoconjugate is of the following formula: [ka] The method is characterized by comprising (wherein Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5), the anti-CD20 antibody is rituximab, the chemotherapeutic agent comprises cyclophosphamide and doxorubicin, and the corticosteroid comprises prednisone.

[0076] In some embodiments, the present invention relates to a method for selecting a therapy for a patient having DLBCL, comprising measuring a macrophage biomarker in a sample from the patient, identifying the patient as one who can benefit from a treatment comprising an immunoconjugate, an anti-CD20 antibody, a chemotherapeutic agent and a corticosteroid, where the amount or level of the macrophage biomarker in the sample is below the amount or level of a reference macrophage biomarker, and the immunoconjugate is of the following formula: [ka] The method is characterized by comprising (wherein Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5), the anti-CD20 antibody is rituximab, the chemotherapeutic agent comprises cyclophosphamide and doxorubicin, and the corticosteroid comprises prednisone.

[0077] In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the amount or level of macrophage biomarkers from the patient is below a reference amount or level of macrophage biomarkers, and the method further includes administering to the patient an effective amount of immunoconjugate, an effective amount of anti-CD20 antibody, an effective amount of chemotherapeutic agent, and an effective amount of corticosteroid.

[0078] In some embodiments, the present invention relates to a method for treating a patient having DLBCL, comprising: (a) measuring a macrophage biomarker in a sample from the patient such that the amount or level of the macrophage biomarker in the sample is below a reference amount or level of macrophage biomarker; and (b) administering to the patient an effective amount of an immunoconjugate, an effective amount of an anti-CD20 antibody, an effective amount of a chemotherapeutic agent, and an effective amount of a corticosteroid, based on the macrophage biomarker measured in step (a), wherein the immunoconjugate has the following formula: [ka] The method is characterized by comprising (wherein Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5), the anti-CD20 antibody is rituximab, the chemotherapeutic agent comprises cyclophosphamide and doxorubicin, and the corticosteroid comprises prednisone.

[0079] In some embodiments, the present invention is a method for treating a patient having DLBCL, comprising administering to the patient an effective amount of an immunoconjugate, an effective amount of an anti-CD20 antibody, an effective amount of a chemotherapeutic agent and an effective amount of a corticosteroid, wherein prior to treatment, it has been determined that the amount or level of a macrophage biomarker in a sample from the patient is below a reference amount or level of a macrophage biomarker, and the immunoconjugate is of the following formula: [ka] The method is characterized by comprising (wherein Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5), the anti-CD20 antibody is rituximab, the chemotherapeutic agent comprises cyclophosphamide and doxorubicin, and the corticosteroid comprises prednisone.

[0080] In some embodiments, the present invention provides a method for treating a patient having DLBCL and in which the amount or level of a macrophage biomarker in a patient sample is below a reference macrophage biomarker amount or level, comprising administering to the patient an effective amount of an immunoconjugate, an effective amount of an anti-CD20 antibody, an effective amount of a chemotherapeutic agent and an effective amount of a corticosteroid, wherein the immunoconjugate has the following formula: [ka] The method is characterized by comprising (wherein Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5), the anti-CD20 antibody is rituximab, the chemotherapeutic agent comprises cyclophosphamide and doxorubicin, and the corticosteroid comprises prednisone.

[0081] In some embodiments of the method that can be combined with any of the aforementioned aspects or embodiments, the immunoconjugate is polatuzumab vedotin.

[0082] In some embodiments of the method, which may be combined with any of the aforementioned aspects or embodiments, the immunoconjugate is administered at a dose of approximately 1.8 mg / kg and rituximab at approximately 375 mg / m². 2 It is administered at a dose of approximately 750 mg / m² of cyclophosphamide. 2 It is administered at a dose of approximately 50 mg / m² of doxorubicin. 2 It is administered in the following doses, with prednisone administered at a dose of approximately 100 mg. [Brief explanation of the drawing]

[0083] [Figure 1]Figure 1 shows a schematic diagram of the POLARIX trial design (clinical trial ID number NCT03274492), a randomized, double-blind trial in previously untreated patients with diffuse large B-cell lymphoma (DLBCL). Pola-R-CHP consists of polatuzumab vedotin (1.8 mg / kg) + rituximab (375 mg / m2), cyclophosphamide (750 mg / m2), doxorubicin (50 mg / m2), and prednisone (100 mg once daily on days 1-5). * = IV on day 1, † = R-CHOP: IV rituximab 375 mg / m2, cyclophosphamide 750 mg / m2, doxorubicin 50 mg / m2 and vincristine 1.4 mg / m2 (maximum 2 mg) on ​​day 1, and oral prednisone 100 mg once daily from days 1 to 5, IPI = International Prognostic Index, ECOG PS = Eastern Cooperative Oncology Group Performance Status, R = Randomization.

[0084] [Figure 2] Figure 2 shows the characteristics of RNA-seq-evaluable patients in the Pola-R-CHP and R-CHOP treatment groups. IPI = International Prognostic Index, n = Number of patients, COO = Cell origin, ABC = Activated B-cell-like, GCB = Germline center B-cell-like.

[0085] [Figure 3A-3B]Figures 3A and 3B show that low M1 macrophage levels are associated with worse progression-free survival (PFS) after R-CHOP treatment, but not after Pola-R-CHP treatment. The PFS probabilities as a function of time (months) for high / low M1 macrophage signatures using QuanTIseq are shown for the R-CHOP (Figure 3A) and Pola-R-CHP (Figure 3B) treatment groups. Insets in Figures 3A and 3B show the PFS hazard ratio (HR) for high M1 macrophage levels versus low M1 macrophage levels in each treatment group. Insets in Figures 3A and 3B also show the estimated 3-year PFS for high and low M1 macrophage levels. The reference group is below the median. HRs were adjusted for IPI score (2 vs. 3-5), age (under 60 vs. over 60), and COO (ABC, GCB, unclassified, unknown). IPI = International Prognostic Index, COO = Cell Origin, ABC = Activated B-cell-like, GCB = Germinal Center B-cell-like

[0086] [Figure 4] Figure 4 shows that lower M1 macrophage levels are associated with lower PFS at 24 months in the R-CHOP treatment group, but not in the Pola-R-CHP treatment group. The proportion of patients within subgroups with progressive disease (PD) by 24 months is shown. The patient subgroups shown are those with low M1 macrophage levels treated with R-CHOP (low R-CHOP), those with high M1 macrophage levels treated with R-CHOP (high R-CHOP), those with low M1 macrophage levels treated with Pola-R-CHP (low Pola-R-CHP), and those with high M1 macrophage levels treated with Pola-R-CHP (high R-CHP). QuanTIseq macrophage M1 levels are expressed as median (p-value by Fisher's exact test). Clopper-Pearson binomial 95% confidence intervals are shown. The M1 macrophage signature distribution was similar between treatment groups.

[0087] [Figure 5A-5B]Figures 5A and 5B show that lower M1 macrophage levels are associated with worse overall survival (OS) after R-CHOP treatment but not after Pola-R-CHP treatment. The OS probability as a function of time (months) for high / low M1 macrophage signatures utilizing QuanTIseq in the R-CHOP (Figure 5A) and Pola-R-CHP (Figure 5B) treatment groups is shown. Insets in Figures 5A and 5B show the OS HR for high M1 macrophage levels versus low M1 macrophage levels in each treatment group. The insets in Figures 5A and 5B also show the 3-year OS estimates for high and low M1 macrophage levels. QuanTIseq macrophage M1 levels are expressed as median values.

[0088] [Figure 6] Figure 6 shows the PFS HR for M1 macrophage levels derived from QuanTIseq and xCell for the Pola-R-CHP and R-CHOP treatment groups. The corresponding forest plots are shown. CI = confidence interval

[0089] [Figure 7] Figure 7 shows baseline patient characteristics from a subset of patients (n=665) from whom transcriptome profiles were generated and analyzed via RNA-seq.

[0090] [Figure 8] Figure 8 shows the progression-free survival (PFS) of patients with high TAM signature versus low TAM signature, ranked by median in the GOYA, MAIN, or POLARIX clinical trials.

[0091] [Figure 9] Figure 9 shows the correlation between TAM signatures and spatially induced macrophage signatures from reactive lymph nodes and DLBCL tumors, using scRNA macrophage data from a publicly available whole lymphoid tissue atlas, as well as the projection of TAM and MacroSig4 signatures onto the corresponding UMAPs.

[0092] [Figure 10] Figure 10 shows the progression-free survival (PFS) of patients with a high TAM signature versus patients with a low TAM signature in the POLARIX trial treated with Pola-R-CHP. [Modes for carrying out the invention]

[0093] I. General Techniques The techniques and procedures described or referenced herein are generally well understood and conventional methodologies by those skilled in the art, such as Sambrook et al., Molecular Cloning: A Laboratory Manual 3d edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Current Protocols in Molecular Biology (FMAusubel et al., eds., (2003)), Methods in Enzymology series (Academic Press, Inc.); PCR 2: A Practical Approach (MJ MacPherson, BD Hames and GR Taylor, eds., (1995)), Harlow and Lane, eds. (1988); Antibodies, A Laboratory Manual, and Animal Cell Culture (RI Freshney, ed., (1987)); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (JE Cellis, ed., 1998) Academic Press. Press, Animal Cell Culture (RIFreshney, ed., 1987), Introduction to Cell and Tissue Culture (JP Mather and PERoberts, 1998) Plenum Press, Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J. B. Griffiths, and D. G. Newell, eds. (1993-8)) J. Wiley and Sons, Handbook of Experimental Immunology (DM Weir and C. C. Blackwell, eds.), Gene Transfer Vectors for Mammalian Cells (J. M. Miller and MPCalos, ed., 1987), PCR: The Polymerase Chain Reaction (Mullis et al., ed., 1994), Current Protocols in Immunology (JEColigan et al., ed., 1991), Short Protocols in Molecular Biology (Wiley and Sons, 1999), Immunobiology (CA Janeway and Sons, 1999), P. Travers, 1997), Antibodies (P. Finch, 1997), Antibodies: A Practical Approach (D. Catty., ed., IRL Press, 1988-1989), Monoclonal Antibodies: A Practical Approach (edited by P. Shepherd and C. Dean, Oxford University Press, 2000), Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999), The Antibodies (M. Zanetti and JD Capra eds., Harwood Academic The widely used methodology described in Publishers, 1995, and Cancer: (V.T. DeVita et al., JBLippincott Company, 1993) is commonly employed.

[0094] II. Definition The aspects and embodiments of the invention described herein are understood to include "a," "consisting of," and "essentially consisting of." Where used herein, the singular forms "a," "an," and "the" include plural references unless otherwise indicated.

[0095] As used herein, the term “about” refers to the normal range of error for each value, as will be readily understood by those skilled in the art. References to “about” values ​​or parameters herein include (and are described) embodiments relating to the value or parameter itself. For example, a description referring to “about X” includes a description of “X.” In some embodiments, “about” may refer to ±15%, ±10%, ±5%, or ±1%, as understood by those skilled in the art.

[0096] As used interchangeably herein, “quantity,” “level,” or “expression level” of a biomarker refers to a level detectable in a biological sample (e.g., a blood sample or a biopsy sample). “Expression” generally refers to the process by which information (e.g., genetic coding information and / or epigenetic information) is converted into a structure that exists and functions within a cell. Therefore, as used herein, “expression” may refer to transcription to polynucleotides, translation to polypeptides, or further to polynucleotide and / or polypeptide modifications (e.g., post-translational modifications of polypeptides). Fragments of transcribed polynucleotides, translated polypeptides, or polynucleotide and / or polypeptide modifications (e.g., post-translational modifications of polypeptides) should also be considered expressed, regardless of whether they originate from transcripts produced by alternative splicing or degraded transcripts, or from post-translational processing of polypeptides, for example, by proteolysis. “Expressed genes” include those transcribed as mRNA into polynucleotides and subsequently translated into polypeptides, and those transcribed into RNA but not translated into polypeptides (e.g., transfer and ribosomal RNAs). Expression levels can be measured by methods known to those skilled in the art and disclosed herein. The expression levels or amounts of biomarkers can be used to identify / characterize subjects with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) that may respond to or benefit from a particular therapy (e.g., a therapy comprising one or more dosing cycles of an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab)).

[0097] The presence and / or expression levels / amounts of the various biomarkers described herein in a sample (e.g., blood sample or biopsy sample) can be analyzed by several methodologies, many of which are known in the art and understood by those skilled in the art, including immunohistochemistry ("IHC"), Western blot analysis, immunoprecipitation, molecular binding assays, ELISA, ELIFA, flow cytometry, fluorescence-activated cell sorting ("FACS"), spatial transcriptomics, spatial proteomics, MassARRAY, proteomics, quantitative blood-based assays (e.g., serum ELISA), biochemical enzyme activity assays, in situ hybridization (ISH), and fluorescence in This includes, but is not limited to, any one of the wide variety of assays that can be performed by situ hybridization (FISH), Southern blotting, Northern blotting, whole-genome sequencing, large-scale parallel DNA sequencing (e.g., next-generation sequencing), NANOSTRING®, polymerase chain reaction (PCR) including quantitative real-time PCR (qRT-PCR) and other amplification-type detection methods for branched DNA, SISBA, TMA, etc., RNA-seq, microarray analysis, gene expression profiling, and / or sequential gene expression analysis ("SAGE"), and protein, gene, and / or tissue array analysis. Typical protocols for evaluating the status of genes and gene products can be found, for example, in Part 2 (Northern blotting), Part 4 (Southern blotting), Part 15 (Immunoblotting), and Part 18 (PCR analysis) of Ausubel et al., eds., 1995, Current Protocols In Molecular Biology. Multiplexed immunoassays, such as those available from Rules Based Medicine or Meso Scale Discovery ("MSD"), may also be used.

[0098] The term “antagonist” is used in its broadest sense and includes any molecule that partially or completely blocks, inhibits, or neutralizes the biological activity of the natural polypeptides disclosed herein. Suitable antagonist molecules include, specifically, antagonist antibodies or antibody fragments (e.g., antigen-binding fragments), fragments or amino acid sequence variants of natural polypeptides, peptides, antisense oligonucleotides, small organic molecules, etc. Methods for identifying polypeptide antagonists may include contacting the polypeptide with a candidate antagonist molecule and measuring a detectable change in one or more biological activities typically associated with that polypeptide.

[0099] The term "anti-CD20 antibody" according to the present invention refers to an antibody capable of binding to CD20 with sufficient affinity so that it is useful as a diagnostic and / or therapeutic agent in targeting CD20. Preferably, the degree of binding of the anti-CD20 antibody to unrelated non-CD20 proteins is less than about 10% of the binding of the antibody to CD20 as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the dissociation constant (Kd value) of the antibody binding to CD20 is ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, or ≤0.1 nM. In certain embodiments, the anti-CD20 antibody binds to an epitope of CD20 that is conserved among CD20 from different species.

[0100] The meanings of "Type I" and "Type II" anti-CD20 antibodies are well known in the art. Generally, anti-CD20 monoclonal antibodies are classified into two different categories based on their mechanism of action in eradicating lymphoma cells. "Type I" anti-CD20 antibodies primarily utilize complement to kill target cells, while "Type II" anti-CD20 antibodies act primarily through a different mechanism called apoptosis. Rituximab (e.g., U.S. Patent No. 5,736,137, which is incorporated entirely herein by reference) and 1F5 are examples of Type I anti-CD20 antibodies, while obinutuzumab (e.g., see International Publication No. 2005 / 044859 and U.S. Patent Application Publication No. 2005 / 0123546, which are incorporated entirely herein by reference) and B1 are examples of Type II antibodies. For example, see Cragg (Blood 103(7), 2004, 2738-2743), Teeling (Blood 104(6), 2004, 1793-1800), European Patent No. 2380910, and International Publication No. 2005 / 044859, the entire contents of which are incorporated herein by reference.

[0101] As used herein, "CD20" refers to the human B lymphocyte antigen CD20 (also known as CD20, B lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5, whose sequence is characterized by SwissProt database entry P11836), a hydrophobic transmembrane protein with a molecular weight of approximately 35 kD located on pre-B lymphocytes and mature B lymphocytes (Valentine, MA, et al., J. Biol. Chem. 264(19) (1989) 11282-11287, Tedder, TF, et al, Proc. Natl. Acad. Sci. USA 85 (1988) 208-12, Stamenkovic, I., et al., J. Exp. Med. 167 (1988) 1975-80, Einfeld, DA et al., EMBO J.7(1988)711-7, Tedder, TF, et al., J.Immunol.142(1989)2560-8). The corresponding human gene is transmembrane 4 domain, subfamily A, member 1, also known as MS4A1. This gene encodes a member of the transmembrane 4A gene family. Members of this nascent protein family are characterized by common structural features and similar intron / exon splice boundaries, and exhibit unique expression patterns between hematopoietic cells and non-lymphoid tissues. This gene encodes a B lymphocyte surface molecule that plays a role in the development and differentiation of B cells into plasma cells. This family member is localized to 11q12 within the cluster of family members. Alternative splicing of this gene results in two transcriptional variants encoding the same protein.

[0102] The terms “CD20” and “CD20 antigen” are used interchangeably herein and include any variant, isoform, and species homolog of human CD20 that is naturally expressed by cells or expressed on cells transfected with the CD20 gene. Binding of the antibodies of the present invention to the CD20 antigen mediates the death of CD20-expressing cells (e.g., tumor cells) by inactivating CD20. The death of CD20-expressing cells may occur by one or more of the following mechanisms: cell death / apoptosis induction, ADCC, and CDC. Synonyms for CD20 recognized in the art include B lymphocyte antigen CD20, B lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5.

[0103] The term "CD20 antigen expression" is intended to indicate a significant level of CD20 antigen expression in cells, such as T cells or B cells. In one embodiment, a patient treated according to the method of the present invention expresses a significant level of CD20 on a B-cell tumor or cancer. A patient having "CD20-expressing cancer" can be determined by standard assays known in the art. For example, CD20 antigen expression is measured by immunohistochemistry (IHC) detection, using FACS, or by PCR-based detection of the corresponding mRNA.

[0104] As used herein, “administer” means a method of giving a certain dose of a compound (e.g., an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab)) or a composition (e.g., a pharmaceutical composition, e.g., a pharmaceutical composition comprising an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab)) to a subject. The compounds and / or compositions used in the methods described herein may be administered, for example, intravenously (e.g., by intravenous injection), subcutaneously, intramuscularly, intradermally, percutaneously, intraarterially, intraperitoneally, intrafocally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, intraperitoneally, subconjunctivally, intravesicularly, transmucosally, intrapericardially, intraumbilically, intraocularly, orally, topically, locally, by inhalation, injection, infusion, continuous infusion, local perfusion directly into target cells, by catheter, lavage, cream, or lipid composition. The method of administration may vary depending on various factors (e.g., the compound or composition to be administered and the severity of the symptom, disease, or disorder being treated).

[0105] In this specification, a “fixed” or “flat” dose of a therapeutic agent (e.g., an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab)) refers to the dose administered to the patient regardless of their body weight or body surface area (BSA). Therefore, this fixed or flat dose is expressed in mg / kg or mg / m². 2 It is specified not as a dosage, but rather as the absolute amount of the therapeutic agent (e.g., mg).

[0106] As used herein, the terms “treatment” or “treating” refer to a clinical intervention designed to alter the natural course of an individual or cells being treated during the course of a clinical lesion. Desired effects of treatment include slowing or reducing the rate of disease progression, recovery or mitigation of the disease state, and remission or improvement of the prognosis. For example, an individual is “treated” if one or more symptoms associated with cancer (e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) are reduced or eliminated, including but not limited to reducing (or destroying) the proliferation of cancer cells, reducing the symptoms resulting from the disease, improving the quality of life of the person with the disease, reducing the dosage of other medications needed to treat the disease, slowing the progression of the disease, and / or extending the survival time of the individual.

[0107] As used herein, “in combination with” or “together with” means administering one treatment regimen together with another. Thus, “in combination with” or “together with” refers to the administration of another treatment regimen before, during, or after the administration of one treatment regimen to an individual. Co-administration may be simultaneous or sequential, preferably, there is a period during which both or all activators exert their biological activity simultaneously. The antibody and the further(s) agents are administered simultaneously or sequentially (e.g., intravenously (iv)), for example, by continuous infusion. When both therapeutic agents are administered sequentially and simultaneously, the doses may be administered in two separate doses on the same day, or one of the agents may be administered on day 1 and the second on days 2-7, preferably days 2-4. Thus, in one embodiment, the term “sequential” means within (approximately) 7 days after administration of the first component, preferably within (approximately) 4 days after administration of the first component, and the term “simultaneously” means at the same time. The term “concurrent administration” with respect to maintenance doses of antibodies and / or additional agents means that maintenance doses can be administered concurrently, for example weekly, or weekly, if the additional agent is administered, for example, every 1 to 3 days and the antibody is administered weekly, provided that the treatment cycle is appropriate for both drugs. Alternatively, the maintenance doses may be administered concurrently sequentially within one day or several days. In preferred embodiments, the anti-CD79b immunoconjugates described herein (e.g., polatuzumab vedotin or its functional equivalent) and anti-CD20 antibodies (e.g., obinutuzumab, rituximab, or their functional equivalents) may be administered in combination with chemotherapy, for example, CHP chemotherapy or a variant of CHP chemotherapy. Thus, in preferred embodiments, the additional chemotherapeutic agent administered concurrently is selected from the group consisting of cyclophosphamide, hydroxydaunorubicin, prednisone or prednisolone and optionally etoposide.

[0108] "Disorder" or "disease" is any condition that would benefit from treatment, including, but not limited to, disorders associated with some degree of abnormal cell proliferation, such as cancer or lymphoma.

[0109] The terms "cancer" and "cancerous" refer to or describe physiological conditions in mammals typically characterized by uncontrolled cell growth. Examples of cancer include, but are not limited to, lymphoma, carcinoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More specific examples of such cancers include, but are not limited to, multiple myeloma and B-cell lymphoma (low-grade / follicular non-Hodgkin lymphoma (NHL), small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small unsevered cell NHL, giant lesion NHL, mantle cell lymphoma, AIDS-associated lymphoma, and Waldenström macroglobulinemia), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), acute myoembolic leukemia (AML), hairy cell leukemia, chronic myeloblastic leukemia (CML), post-transplant lymphoproliferative disorder (PTLD), and myelodysplastic syndrome (MDS), as well as associated metastases. In some embodiments, the cancer is a lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). In some embodiments, the lymphoma is a low-grade lymphoma. In some embodiments, the lymphoma is a B-cell lymphoma. In some embodiments, the B-cell lymphoma is germinal center-derived B-cell lymphoma. In some embodiments, the B-cell lymphoma is NHL. In some embodiments, the lymphoma is diffuse large B-cell lymphoma (DLBCL). In some embodiments, the lymphoma is DLBCL. In some embodiments, the DLBCL is a germinal center B-cell-like (GCB) cell-derived subgroup or an activated B-cell-like (ABC) cell-derived subgroup of DLBCL. In some embodiments, the lymphoma is a CD20-positive lymphoma.

[0110] The term “tumor” refers to the growth and proliferation of all neoplastic cells, whether malignant or benign, as well as all precancerous and cancerous cells and tissues. The terms “cancer,” “cancerous,” “proliferative disorder,” “proliferative disorder,” and “tumor” are not mutually exclusive as used herein.

[0111] As used herein, “metastasis” means the spread of cancer (e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) from its primary site to other parts of the body. Cancer cells may detach from the primary tumor, infiltrate the lymphatic vessels and blood vessels, circulate through the bloodstream, and grow (metastasize) in distal lesions within normal tissues at other parts of the body. Metastasis can be local or distal. Metastasis is a sequential process in which tumor cells detach from the primary tumor, migrate through the bloodstream, and halt at a distal site. At the new site, cells can establish a blood supply, grow, and form life-threatening masses. Both stimulative and inhibitory molecular pathways within tumor cells control this behavior, and interactions between tumor cells and host cells at the distal site are also important.

[0112] The term "anti-cancer therapy" refers to therapies useful for treating cancer (e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). Examples of anti-cancer agents include, but are not limited to, immunomodulators or agents that increase or activate one or more immune costimulatory receptors, chemotherapeutic agents, proliferation inhibitors, cytotoxic agents, agents used in radiotherapy, anti-angiogenic agents, apoptotic agents, antitubulin agents, and other agents for treating cancer. Combinations of these are also included in the present invention. In some embodiments, anticancer therapies include cyclophosphamide, doxorubicin, and prednisone (CHP) or variations thereof (e.g., CHOEP chemotherapy, CHOP-14 chemotherapy, or ACVBP chemotherapy (e.g., Examples and European Patent No. 2380910, International Publication No. 2005 / 044859, and Scott, 2014 and 2015, loc. cit.)).

[0113] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or interferes with cellular function and / or causes cell death or cell destruction. Examples of cytotoxic agents include radioactive isotopes (e.g., At). 211 , I 131 , I 125 , Y 90 Re 186 Re 188 Sm 153 , Bi 212 , P 32 Pb 212 Examples of antitumor agents or anticancer agents include, but are not limited to, the following: , and radioisotopes of Lu; chemotherapeutic agents or drugs (e.g., methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin, or other inserts); growth inhibitors; enzymes and their fragments, e.g., nucleases; antibiotics; toxins (including their fragments and / or variants), such as low-molecular-weight toxins or enzymatically active toxins of bacterial, fungal, plant, or animal origin; and various antitumor agents or anticancer agents disclosed below.

[0114] "Chemotherapy agents" include chemical compounds useful for treating cancer. Examples of chemotherapy agents include erlotinib (TARCEVA®, Genentech / OSI Pharm.) and bortezomib (VELCADE®, Millennium). Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant (FASLODEX®, AstraZeneca), sunitib (SUTENT®, Pfizer / SUGAIN), letrozole (FEMARA®, Novartis), imatinib mesylate (GLEEVEC®, Novartis), finasnate (VATALANIB®, Novartis), oxaliplatin (ELOXATIN®, Sanofi), 5-FU (5-fluorouracil), leucovorin, rapamycin (sirolimus, RAPAMUNE®, Wyeth), lapatinib (TYKERB®, GSK572016, Glaxo Smith) Alkylating agents such as Kline, ronafamib (SCH 66336), sorafenib (NEXAVAR®, Bayer Labs), gefitinib (IRESSA®, AstraZeneca), AG1478, thiotepa and CYTOXAN® cyclophosphamide, alkyl sulfonates such as busulfan, improsulfan and pigosulfan, aziridines such as benzodopa, carbocone, metredopa, uredopa, altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and ethyleneimines and methylamelamamine including trimethylomellamine, acetone Genin (especially bratacin and bratacinone), camptothecin (including topotecan and irinotecan), bryostatin, calistatin, CC-1065 (including synthetic analogs of adzeresin, karzeresin, and bizeresin), cryptophycin (especially cryptophycin 1 and cryptophycin 8), corticosteroids (including prednisone and prednisolone), cyproterone acetate, 5-reductase (including finasteride and dutasteride), vorinostat, romidepsin, panobinostat, valproic acid,Mosetinostat, dorastatin, aldesleukin, talc duocalmycin (including synthetic analogs KW-2189 and CB1-TM1), eleuterobin, pancratistatin, sarcodicin, spongstatin, chlorambucil, chromafazine, chlorophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobenbitin, fenestrine, prednimustine, trophosphamide, nitrogen mustards such as uracil mustard, nitrosourea such as carmustine, chlorozotosine, fotemustine, lomustine, nimustine, and ranimustine, engine antibiotics (e.g., calicheamicin, especially calicheamicin γ1I and calicheamicin ω1I (Angew Chem. Intl. Ed. Engl. 1994) 33:183-186), antibiotics such as dynemycin A, bisphosphonates such as clodronate, esperamycin, and neocartinostatin chromophores and related pigment proteins (endiin antibiotic chromophores), acrasinomycin, actinomycin, ausuramycin, azaserin, bleomycin, kactinomycin, carabicin, caminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detrevicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin, morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epi Antimetabolites such as rubicin, esorubicin, idarubicin, marcelomycin, mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, porphyromycin, puromycin, queramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zorubicin, methotrexate, and 5-fluorouracil (5-FU), folic acid analogs such as denopterin, methotrexate, pteropterin, and trimethrexate, purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine, ancitabine, azacitidine, 6-azauridine, carmoflu, cytarabine, dideoxyuridine,Pyrimidine analogs such as doxifluridine, enocitabine, and phloxuridine; androgens such as carsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testactone; anti-adrenaline such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as floric acid; acegraton, aldofsphamide glycoside, aminolevulinic acid, enyluracil, amsacrin, bestrabusil, bisantren, edatraxate, defoliant Famin, demecorsin, diazicone, erhomitin, eriptinium acetate, epotilon, etoglucide, gallium nitrate, hydroxyurea, lentinan, ronidynin, mayansinoids such as mayansin and anthamitosin, mitoglucon, mitoxantrone, mopidamnol, nitrrelin, pentostatin, fenamet, pirarubicin, losoxantrone, podophyllic acid, 2-ethylhydrazide, procarbazine, PSK (registered trademark) polysaccharide complex (JHS Natural Products (Eugene, Oreg.), Lazoxane, Rhizoxin, Schizofuran, Spirogermanium, Tenuazonic Acid, Triadicone, 2,2',2"-Trichlorotriethylamine, Trichothecenes (especially T-2 toxin, Beraclin A, Loridine A, Angidin), Urethane, Vindesine, Dacarbazine, Mannomustine, Mitobronitol, Mitractol, Pipobroman, Gacitosine, Arabinoside ("Ara-C"), Cyclophosphamide, Thiotepa, Taxoids, e.g., TAXOL (Paclitaxel, Bristol-Myers Squibb Oncology, Princeton, New Jersey), ABRAXANE (Registered Trademark) (Cremophor-free), Albumin-Modified Nanoparticle Formulations of Paclitaxel (American Pharmaceutical Partners (Schaumburg, Illinois), and TAXOTERE® (docetaxel, doxetaxel, Sanofi-Aventis), chlorambucil, GEMZAR® (gemcitabine), 6-thioguanine, mercaptopurine, methotrexate, platinum analogs such as cisplatin and carboplatin, vinblastine, etoposide (VP-16), ifosfamide, mitoxantrone, vincristine, NAVELBINE® (vinorelbine),Examples include novantrone, teniposide, edatrexate, daunomycin, aminopterin, capecitabine (XELODA®), ibandronate, CPT-11, topoisomerase inhibitor RFS2000, difluoromethylornithine (DMFO), retinoids such as retinoic acid, and any of the above pharmaceutically acceptable salts, acids, and derivatives. α,

[0115] Other chemotherapeutic agents include (i) anti-hormone agents that act to modulate or inhibit the hormonal effects on tumors, such as tamoxifen (NOLVADEX®, tamoxifen citrate, etc.), anti-estrogen and selective estrogen receptor modulators (SERMs), such as raloxifene, droloxifene, iodoxifene, 4-hydroxytamoxifen, trioxyfen, keoxyfen, LY117018, onapristone, and FARESTON® (toremifine citrate), and (ii) aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production in the adrenal gland, such as 4(5)-imidazole, and Minoglutethimide, MEGASE® (Megestrol acetate), AROMASIN® (Exemestane, Pfizer), Formestany, Fadrozol, RIVISOR® (Vorozol), FEMARA® (Letrozole, Novartis), and ARIMIIDEX® (Anastrozole, AstraZeneca), (iii) antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin, buserelin, trypterine, medroxyprogesterone acetate, diethylstilbestrol, Premarin, fluoxymesterone, total translate acids, fenretinide, and troxacitabine (1,(iv) Protein kinase inhibitors (e.g., anaplastic lymphoma kinase (Alk) inhibitors, e.g., AF-802 (also known as CH-5424802 or alectinib)), (v) Lipid kinase inhibitors, (vi) Antisense oligonucleotides, in particular those that inhibit gene expression in signaling pathways involved in abnormal cell proliferation, e.g., PKC-alpha, Ralf, and H-Ras, (vii) Ribozymes, e.g., VEGF expression inhibitors (e.g., ANGIOZYME®) and HER2 expression inhibitors, (viii) Vaccines such as gene therapy vaccines, e.g., ALLOVECTIN®, LEUVECTIN®, and VAXID®, PROLEUKIN®, rIL-2, topoisomerase 1 inhibitors such as LURTOTECAN®, ABARELIX® rmRH, and (ix) pharmaceutically acceptable salts, acids, and derivatives of any of the above.

[0116] Examples of chemotherapy agents include antibodies such as alemtuzumab (Campath), bevacizumab (AVASTIN®, Genentech), cetuximab (ERBITUX®, Imclone), panitumumab (VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech / Biogen Idec), pertuzumab (OMNITARG®, 2C4, Genentech), trastuzumab (HERCEPTIN®, Genentech), tositumomab (Bexxar, Corixia), and antibody-drug conjugates such as gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Additional humanized monoclonal antibodies that have therapeutic potential as a drug in combination with the listed compounds include apolizumab, aselizumab, atrizumab, bapinuzumab, vibatuzumab meltansine, cantuzumab meltansine, sedelizumab, cerorizumab pegol, sidofcituzumab, sidotuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erulizumab, felbizumab, fontrizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, rabetuzumab, lintuzumab, matzumab, mepolizumab, motabizumab, motobizumab, and na Talizumab, nimotuzumab, norovizumab, numabizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pecufcituzumab, pertuzumab, paxerizumab, larivizumab, ranibizumab, reslivizumab, reslizumab, reslivizumab, reslivizumab, loberizumab, lupizumab, sibrotuzumab, cyprizumab, sontuzumab, takatatuzumab, tetraxetan, tadoxizumab, talizumab, tefibazumab, tocilizumab, tralizumab, tucotsuzumab, cermoleukin, tucucituzumab, umavizumab, urtoxazumab, ustekinumab, vizilizumab, and interleukin-12 One example is the anti-interleukin-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), a full-length IgG1λ antibody consisting solely of human sequences that has been genetically modified to recognize the p40 protein.

[0117] Chemotherapy agents also include "EGFR inhibitors," which refer to compounds that bind to EGFR or otherwise directly interact with it, inhibiting or reducing its signaling activity, and are alternatively called "EGFR antagonists." Examples of such drugs include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB 8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see U.S. Patent No. 4,943,533 by Mendelsohn et al.) and their variants, e.g., chimeric 225 (C225 or cetuximab, ERBUTIX®) and reshaped human 225 (H225) (International Publication No. 96 / 40210, Imclone Systems). See Inc.), IMC-11F8, fully human EGFR-targeted antibody (Imclone), antibody that binds to type II mutant EGFR (U.S. Patent No. 5,212,290), humanized antibodies and chimeric antibodies that bind to EGFR as described in U.S. Patent No. 5,891,996, and human antibodies that bind to EGFR, such as ABX-EGF or panitumumab (see International Publication No. 98 / 50433, Abgenix / Amgen), EMD 55900 (Stragliotto et al. Eur.J. Cancer) Examples include 32A:636-640 (1996), EMD7200 (matsuzumab), humanized EGFR antibody against EGFR that competes with both EGF and TGF-alpha for EGFR binding (EMD / Merck), human EGFR antibody, HuMax-EGFR (GenMab), known as E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 and E7.6.3, a fully human antibody described in U.S. Patent No. 6,235,883, MDX-447 (Medarex Inc.), and mAb 806 or humanized mAb 806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)).Anti-EGFR antibodies can be conjugated with cytotoxic agents to generate immunoconjugates (see, for example, European Patent Application Publication No. 659,439A2, Merck Patent GmbH). Examples of EGFR antagonists include U.S. Patents Nos. 5,616,582, 5,457,105, 5,475,001, 5,654,307, 5,679,683, 6,084,095, 6,265,410, 6,455,534, 6,521,620, 6,596,726, 6,713,484, 5,770,599, 6,140,332, and 5,866. Examples include small molecules such as compounds described in publications 572, 6399, 602, 6344, 459, 6602, 863, 6391, 874, 6344, 455, 5760, 041, 6002, 008, and 5747, 498, as well as the following PCT publications: International Publication Nos. 98 / 14451, 98 / 50038, 99 / 09016, and 99 / 24037.Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, TARCEVA®, Genentech / OSI Pharmaceuticals), PD183805 (CI1033, 2-propenamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-dihydrochloride, Pfizer) AstraZeneca Inc., ZD1839, Gefitinib (IRESSA®) 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, ZM105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca), BIBX-1382 (N8-(3-chloro-4-fluorophenyl)-N2-(1-methyl-piperidine-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim), PKI-166((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidine-6-yl]phenol), (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine), CL-387785(N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butinamide), EKB-569(N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano- Examples include 7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butinamide) (Wyeth), AG1478 (Pfizer), AG1571 (SU5271, Pfizer), and dual EGFR / HER2 tyrosine kinase inhibitors, such as lapatinib (TYKERB®, GSK572016, or N-3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine).

[0118] As chemotherapeutic agents, there are also "tyrosine kinase inhibitors" such as EGFR-targeted drugs mentioned in the previous paragraph, insulin receptor tyrosine kinase inhibitors such as anaplastic lymphoma kinase (ALK) inhibitors, for example AF-802 (also known as CH-5424802 or alectinib), ASP3026, X396, LDK378, AP26113, crizotinib (XALKORI®) and ceritinib (ZYKADIA®), small molecule HER2 tyrosine kinase inhibitors such as TAK165 available from Takeda, and ErbB2 receptor tyrosine kinase inhibitors. Dual HER inhibitors such as CP-724 and 714 (Pfizer and OSI), which are oral selective inhibitors of HER2; EKB-569 (available from Wyeth), which preferentially binds to EGFR but inhibits both HER2 and EGFR overexpressing cells; lapatinib (GSK572016, available from Glaxo-SmithKline), an oral HER2 and EGFR tyrosine kinase inhibitor; pan-HER inhibitors such as PKI-166 (available from Novartis) and canertinib (CI-1033, Pharmacia); and ISIS, which inhibits Raf-1 signaling. Raf-1 inhibitors such as the antisense agent ISIS-5132 available from Pharmaceuticals, non-HER-targeted TK inhibitors such as imatinib mesylate (GLEEVEC®, available from Glaxo SmithKline), multi-target tyrosine kinase inhibitors such as sunitinib (SUTENT®, available from Pfizer), VEGF receptor tyrosine kinase inhibitors such as Vatalanib (PTK787 / ZK222584, available from Novartis / Schering AG), MAPK extracellular regulatory kinase I inhibitor CI-1040 (available from Pharmacia), quinazolines, e.g., PD 153035, 4-(3-chloroanilino)quinazoline, pyridopyrimidine, pyridopyrimidine, CGP 59326, CGP 60261 and CGP Pyrrolopyrimidines such as 62706, pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidines, curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide), tyrofostine containing the nitrothiophene moiety, PD-0183805 (Warner-Lambert), antisense molecules (e.g., those that bind to HER coding nucleic acids), quinoxaline (US Patent No. 5,804,396), triphostine (US Patent No. 5,804,396), pan-HER inhibitors such as ZD6474 (AstraZeneca), PTK-787 (Novartis / Schering AG), CI-1033 (Pfizer), Affinitac (ISIS 3521, Isis / Lilly), imatinib mesylate (GLEEVEC®), PKI 166 (Novartis), GW2016 (Glaxo SmithKline), CI-1033 (Pfizer), EKB-569 (Wyeth), Semaxinib (Pfizer), ZD6474 (AstraZeneca), PTK-787 (Novartis / Schering AG), INC-1C11 (Imclone), Rapamycin (Sirolimus, RAPAMUNE®), or the following patent publications: U.S. Patent No. 5,804,396, International Publication No. 1999 / 09016 (American Cyanamid), International Publication No. 1998 / 43960 (American Cyanamid), International Publication No. 1997 / 38983 (Warner Lambert), International Publication No. 1999 / 06378 (Warner Lambert), International Publication No. 1999 / 06396 (Warner Lambert) Examples include those listed in Lambert, Pfizer, Inc. No. 1996 / 30347, Zeneca No. 1996 / 33978, Zeneca No. 1996 / 3397, and Zeneca No. 1996 / 33980.

[0119] Chemotherapy agents include dexamethasone, interferon, colchicine, methoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostin, arsenic trioxide, asparaginase, BCG vaccine, bevacizumab, besarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, erlotinib, filgrastim, histrelin acetate, ibritumomab, interferon alfa-2a, and interferon alfa. -2b, lenalidomide, levamizole, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelbequin, parifermin, pamidronate, pegademase, pegasparagase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium, quinacrine, rasburicase, salglamostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, ATRA, barrubicin, zoledronate, and zoledronic acid, as well as pharmaceutically acceptable salts thereof.

[0120] Other chemotherapy agents include hydrocortisone, hydrocortisone acetate, cortisone acetate, thixocortol pivalate, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, flucortolone, and hydrocortisone-17-butyrate. Hydrocortisone-17-valerate, acromethasone dipropionate, betamethasone valerate, betamethasone dipropionate, prezicalvert, clobetazone-17-butyrate, clobetasol-17-propionate, flucorlon caproate, flucorlon pivalate and flupredniden acetate, phenylalanine-glutamine-glycine (FEG) and its D-isomer (feG) (IMULAN Immunoselective anti-inflammatory peptides (ImSAIDs) such as BioTherapeutics, LLC; antirheumatic drugs such as azathioprine, cyclosporine (cyclosporine A), D-penicillamine, gold salt, hydroxychloroquine, leflunomideminocycline, and sulfasalazine; tumor necrosis factor α (TNFα) blockers such as etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol (Cimzia), and golimumab (Simponi); interleukin-1 (IL-1) blockers such as anakinra (Kineret); T-cell costimulatory blockers such as abatacept (Orencia); tocilizumab (ACTEME) Interleukin-6 (IL-6) blockers such as RA(registered trademark), interleukin-13 (IL-13) blockers such as lebrikizumab, interferon-alpha (IFN) blockers such as lontalizumab, beta-7 integrin blockers such as rhuMAb beta-7, IgE pathway blockers such as anti-M1 prime, secreted homotrimer LTa3 and membrane-bound heterotrimer LTa1 / β2 blockers, such as anti-lymphotoxin α (LTa), radioisotopes (e.g., At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212 and Lu radioisotopes), various investigational drugs, such as thioplatin, PS-341, phenylbutyrate,ET-18-OCH3, or farnesyltransferase inhibitors (L-739749, L-744832), polyphenols such as quercetin, resveratrol, piceatannol, epigallocatechin gallate, theaflavin, flavanol, procyanidin, betulinic acid and its derivatives, autophagy inhibitors such as chloroquine, delta-9-tetrahydrocannabinol (dronabinol, MARINOL®), beta-rapacone, lapachol, colchicine, betulinic acid, acetylcamptothecin, scopolectin, and 9-aminocamptothecin, podophyllotoxin, tegafur (UFTORAL®), bexarotene (TARGRETIN®), clodronate (e.g., BONEFOS® or OSTAC®), etidronate (D Bisphosphonates such as IDROCAL(registered trademark), NE-58095, zoledronic acid / zoledronate (ZOMETA(registered trademark)), alendronate (FOSAMAX(registered trademark)), pamidronate (AREDIA(registered trademark)), tildronate (SKELID(registered trademark)), or risedronate (ACTONEL(registered trademark)), and epidermal growth factor receptor (EGF-R), vaccines such as THERATOPE(registered trademark) vaccine, perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS341), CCI-779, tipifarnib (R11577), olafenib, ABT510, Bcl-2 inhibitors such as oblimersen sodium (GENASENSE(registered trademark)), pixantrone, ronfarnib (SCH Examples include farnesyltransferase inhibitors such as 6636 (SARASARTM), any of the above pharmaceutically acceptable salts, acids, or derivatives, as well as combinations of two or more of the above, such as CHP, an abbreviation for combination therapy of cyclophosphamide, doxorubicin, and prednisolone (prednisone), and FOLFOX, an abbreviation for treatment regimens with oxaliplatin (ELOXATIN®) in combination with 5-FU and leucovorin.

[0121] Other chemotherapeutic agents may include nonsteroidal anti-inflammatory drugs (NSAIDs) that have analgesic, antipyretic, and anti-inflammatory effects. Examples of NSAIDs include non-selective inhibitors of the enzyme cyclooxygenase. Specific examples of NSAIDs include aspirin, propionic acid derivatives such as ibuprofen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin, and naproxen, acetic acid derivatives such as indomethacin, sulindac, etodolac, and diclofenac, enolic acid derivatives such as piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, and isoxicam, fenamic acid derivatives such as mefenamic acid, meclofenamic acid, flufenamic acid, and tolfenamic acid, and COX-2 inhibitors such as celecoxib, etoricoxib, lumiracoxib, parecoxib, rofecoxib, and valdecoxib. NSAIDs may be indicated for the relief of symptoms of conditions such as rheumatoid arthritis, osteoarthritis, inflammatory arthropathy, ankylosing spondylosis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, metastatic bone pain, headache, and migraine, postoperative pain, mild to moderate pain resulting from inflammation and tissue injury, fever, bowel obstruction, and renal colic.

[0122] The “effective dose” of a compound, e.g., an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) or a composition thereof (e.g., a pharmaceutical composition) is at least the minimum amount required to achieve a desired therapeutic outcome, e.g., a measurable increase in overall survival or progression-free survival for a particular disease or disorder (e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). The effective dose as used herein may vary depending on factors such as the patient’s disease status, age, sex, and weight, as well as the antibody’s ability to induce the desired response in the subject. The effective dose is also the amount at which the therapeutically beneficial effect outweighs any toxic or adverse effects of the treatment. Beneficial or desired outcomes for prophylactic use include the elimination or reduction of risk, reduction of severity, or delay of disease onset, including the biochemical, histological, and / or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes that appear during the onset of the disease. For treatment use, beneficial or desired outcomes include clinical outcomes such as reduction of one or more symptoms attributable to the disease (EORTC QLQ-C30, e.g., fatigue, nausea, vomiting, pain, dyspnea, insomnia, loss of appetite, constipation, diarrhea, or general level of physical, emotional, cognitive, or social functioning), improvement from baseline in the FACT-Lym functional assessment of cancer-treated lymphoma subscale score, reduction in the dose of other medications required to treat the disease, enhancement of the effect of another medication by targeting, etc., delay of disease progression (e.g., progression-free survival), delay of clear clinical progression (e.g., progression of cancer-related pain), deterioration of the East Coast Clinical Oncology Group (ECOG) performance status (PS) (e.g., how the disease affects the patient's ability to perform daily activities), and / or initiation of subsequent systemic anticancer therapy, and / or extension of overall survival.In the case of cancer or tumors, an effective dose of a drug may have the effect of reducing the number of cancer cells, reducing tumor size, inhibiting (i.e., delaying to some extent, or preferably stopping) the invasion of cancer cells into peripheral organs, inhibiting (i.e., delaying to some extent, or preferably stopping) tumor metastasis, inhibiting tumor growth to some extent, and / or alleviating to some extent one or more of the symptoms associated with the disorder. An effective dose may be administered in one or more doses. In this invention, an effective dose of a drug, compound, or pharmaceutical composition is an amount sufficient to directly or indirectly achieve a prophylactic or treatment. As understood in the clinical field, an effective dose of a drug, compound, or pharmaceutical composition may or may not be achieved in combination with another drug, compound, or pharmaceutical composition. Thus, “effective dose” may be considered in relation to the administration of one or more therapeutic agents, and a monotherapy agent may be considered to be given in an effective dose if, when combined with one or more other agents, the desired result can or does not occur.

[0123] "Immunogenicity" refers to the ability of a particular substance to trigger an immune response. Tumors are immunogenic, and enhancing tumor immunogenicity helps the immune response to clear tumor cells. Examples of enhancing tumor immunogenicity include, but are not limited to, treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

[0124] "Individual response" or "response" can be evaluated using any endpoint demonstrating a benefit to the subject, including, but not limited to, (1) some degree of inhibition of disease progression (e.g., progression of cancer, e.g., lymphoma) (including slowing of cessation and complete cessation), (2) reduction in tumor size, (3) suppression of cancer cell invasion into adjacent peripheral organs and / or tissues (i.e., reduction, slowing or complete cessation), (4) inhibition of metastasis (i.e., reduction, slowing or complete cessation), (5) some degree of relief of one or more symptoms associated with the disease or disability (e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)), (6) increased or prolonged survival, including overall survival and progression-free survival, and / or (9) a reduction in mortality at a given time after treatment.

[0125] The terms "effective response" or "responsiveness" and similar phrases to a treatment using a pharmaceutical agent refer to a clinical or treatment benefit bestowed upon a subject who is at risk of or suffering from a disease or disorder such as cancer. In one embodiment, such benefit may include one or more of the following: extension of survival (including overall survival and progression-free survival), achieving an objective response (including complete or partial response), or improving the signs or symptoms of cancer.

[0126] A patient who "does not show an effective response" to the treatment is a patient who does not experience any of the following: an extension of survival time (including overall survival and progression-free survival), an objective response (including complete or partial response), or an improvement in signs or symptoms of cancer.

[0127] As used herein, the term “survival” means the period during which a patient is alive, and includes overall survival and progression-free survival.

[0128] As used herein, “overall survival” (OS) refers to the time from enrollment in the study to death from any cause. As used herein, “overall survival rate” refers to the proportion of subjects who are alive at a specific time, for example, 6 months, 1 year, or 5 years from the time of diagnosis or treatment.

[0129] As used herein, “complete response” or “CR” means the disappearance of all evidence of the disease.

[0130] As used herein, “partial response” or “PR” means a measurable reduction in symptoms, a reduction in the direct or indirect pathological consequences of lymphoma, a decrease in the rate of disease progression, improvement or mitigation of the disease state, or prevention of metastasis without ruling out all evidence of the disease.

[0131] As used herein, “progression-free survival” (PFS) refers to the length of time during and after treatment in which the treated disease (e.g., cancer, e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) does not worsen (e.g., progression of lymphoma or death as a result of any cause). PFS may include the time in which the patient experiences a complete or partial response and the time in which the patient experiences stable disease.

[0132] As used herein, “extend survival” means increasing overall survival or progression-free survival in treated patients compared to untreated patients (e.g., compared to patients untreated with a drug), or compared to patients who do not express a specified level of biomarker, and / or compared to patients treated with an approved antitumor agent. Objective response means a measurable response, including complete response (CR) or partial response (PR).

[0133] Those skilled in the art can easily determine whether a given clinical outcome is improved according to the present invention (for example, compared to treatment without the use of an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab)). For example, “improvement” in this context means that the clinical outcome (resulting from treatment with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin or its functional equivalent) and an anti-CD20 antibody (e.g., obinutuzumab / obinutuzumab or rituximab functional equivalent) in particular combination with chemotherapy, particularly in combination with CHP chemotherapy) is improved by the use of an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) in particular combination with chemotherapy, particularly in combination with CHP chemotherapy. This means that the clinical outcomes are at least 3% higher, at least 5% higher, at least 7% higher, at least 10% higher, at least 15% higher, at least 20% higher, at least 25% higher, at least 30% higher, at least 40% higher, at least 50% higher, at least 75% higher, at least 100% higher, or at least 120% higher compared to equivalent treatment without the use of ) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0134] The time at which clinical outcomes / clinical endpoints are assessed can be readily determined by those skilled in the art. In principle, it is determined when the difference in clinical outcomes / clinical endpoints between the two treatments (e.g., polatuzumab vedotin and rituximab (or obinutuzumab) treatment versus rituximab (or obinutuzumab) treatment) becomes apparent. This time could be, for example, at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 12 months, at least 18 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, or at least 48 months after the start of treatment.

[0135] As used herein, “delaying the progression” of a disorder or disease means delaying, interfering with, slowing, stabilizing, and / or postponing the progression of a disease or disorder (e.g., cancer, e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). This delay may be of varying durations depending on the disease history and / or the subject being treated. As will be apparent to those skilled in the art, sufficient or significant delay may substantially encompass prevention in that the subject does not develop the disease.

[0136] As used herein, the term "reduce or inhibit cancer recurrence" means reducing or inhibiting the recurrence of a tumor or cancer, or the progression of a tumor or cancer.

[0137] "To reduce or inhibit" means the ability to result in an overall reduction of 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or greater. Reducing or inhibiting may refer to the symptoms, presence or size of metastases, or size of the primary tumor of the disorder being treated (e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)).

[0138] As used herein, “subject” or “individual” means a mammal, including but not limited to humans or non-human mammals such as cattle, horses, dogs, sheep, or cats. In some embodiments, the subject is human. The patient is also human as used herein.

[0139] The terms “detect” and “detect” are used most broadly herein to include both qualitative and quantitative measurements of a target molecule. Detection includes simply identifying the presence of a target molecule in a sample and determining whether the target molecule is present in the sample at a detectable level. Detection may be direct or indirect.

[0140] "Tumor-infiltrating immune cells," as used herein, refers to any immune cells present in a tumor or a sample thereof. Tumor-infiltrating immune cells include, but are not limited to, intratumor immune cells, peritumor immune cells, other tumor stromal cells (e.g., fibroblasts), or any combination thereof. Such tumor-infiltrating immune cells may include, for example, macrophages (e.g., M1 macrophages, tumor-associated macrophages, or M2 macrophages), monocytes, T lymphocytes (e.g., CD8+ T lymphocytes and / or CD4+ T lymphocytes), B lymphocytes, or granulocytes (e.g., neutrophils, eosinophils, and basophils), dendritic cells (e.g., finger-entrapment dendritic cells), histiocytes, and other myeloid cells, including natural killer cells.

[0141] As used herein, the term “biomarker” refers to an indicator, e.g., predictive, diagnostic, and / or prognostic, that can be detected in a sample (e.g., tumor tissue sample (e.g., lymphoma tumor tissue sample, e.g., B-cell lymphoma tumor tissue sample, e.g., non-Hodgkin lymphoma tumor tissue sample, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma) tumor tissue sample), blood sample, or biopsy sample). A biomarker may serve as an indicator of a specific subtype of a disease or disorder characterized by specific molecular, pathological, histological, and / or clinical features (e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). In some embodiments, a biomarker is a gene (e.g., any of the genes described herein). Examples of biomarkers include, but are not limited to, polypeptides, polynucleotides (e.g., DNA, and / or RNA), polynucleotide copy number alterations (e.g., DNA copy number), polypeptide and polynucleotide modifications (e.g., post-translational modifications), carbohydrates, and / or glycolipid-based molecular markers. In some embodiments, the biomarker is gene expression levels. In some embodiments, the biomarker is the M1 macrophage gene signature set score. In some embodiments, the biomarker is the tumor-associated macrophage (TAM) gene signature set score. In some embodiments, the biomarker is cells (e.g., immune cells, e.g., macrophages, e.g., M1 macrophages or M2 macrophages). In some embodiments, the biomarker is the quantity of M1 macrophages. In some embodiments, the biomarker is the quantity of tumor-associated macrophages.

[0142] As used herein, the term “macrophage biomarker” refers to a biomarker that indicates the quantity, level, characteristics, or phenotype of macrophages in a sample (e.g., tumor tissue sample (e.g., lymphoma tumor tissue sample, e.g., B-cell lymphoma tumor tissue sample, e.g., non-Hodgkin lymphoma tumor tissue sample, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma) tumor tissue sample), blood sample, or biopsy sample). In some embodiments, the macrophage biomarker is a gene (e.g., any of the genes described herein). In some embodiments, the macrophage biomarker is a polypeptide, polynucleotide (e.g., DNA, and / or RNA), polynucleotide copy number modification (e.g., DNA copy number), polypeptide and polynucleotide modification (e.g., post-translational modification), carbohydrate, and / or glycolipid-based molecular marker. In some embodiments, the macrophage biomarker is a gene expression value that may reflect one or more genes (e.g., one or more genes described herein). In some embodiments, the macrophage biomarker is the M1 macrophage gene signature set score. In some embodiments, the macrophage biomarker is the tumor-associated macrophage (TAM) gene signature set score. In some embodiments, the macrophage biomarker is a cell (e.g., an M1 macrophage, a tumor-associated macrophage). In some embodiments, the macrophage biomarker is a certain amount of M1 macrophages. In some embodiments, the macrophage biomarker is a certain amount of tumor-associated macrophages.

[0143] A biomarker is “predictive” according to the Invention if it can be used (in combination of optionally one or more biomarkers) to identify patients who will respond to treatment with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) (especially in combination with chemotherapy, especially in combination with CHP chemotherapy). In some embodiments, a biomarker is predictive if the treatment response differs among subgroups of patients defined by the biomarker. In this regard, it is preferable that the predictive biomarker is a biomarker as defined elsewhere in this Invention. A particular example of a predictive biomarker evaluated in the context of the Invention is the macrophage biomarker described herein.

[0144] The term "antibody" includes monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region), antibody compositions having polyepitope specificity, and multispecific antibodies (e.g., bispecific antibodies, diabodies, and single-chain molecules, as well as antibody fragments containing antigen-binding fragments such as Fab, F(ab')2, and Fv). The term "immunoglobulin" (Ig) is used interchangeably with "antibody" herein.

[0145] The basic four-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light chains (L) and two identical heavy chains (H). IgM antibodies consist of five basic heterotetrameric units plus an additional polypeptide called a J chain, containing 10 antigen-binding sites. IgA antibodies, on the other hand, are composed of 2 to 5 basic four-chain units, which can polymerize to form multivalent aggregates with the J chain. In the case of IgG, the four-chain unit is generally about 150,000 daltons. Each L chain is linked to the H chain by one disulfide covalent bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Both the H and L chains also have regularly spaced intrachain disulfide crosslinks. Each H chain has a variable domain (V) at its N-terminus.H ) has, and subsequently each of the α and γ chains has three constant domains (C H ), and for μ and ε isotypes, four C H It has a domain. Each L chain has a variable domain (V) at its N-terminus. L It has a constant domain at the opposite end. L V H They are aligned as C L This is the first constant domain of the heavy chain (C H 1) is aligned with the following. Certain amino acid residues are thought to form an interface between the light chain variable domain and the heavy chain variable domain. V H and V L The two molecules pair together to form a single antigen-binding site. For the structures and properties of various classes of antibodies, see, for example, page 71 and Chapter 6 of Basic and Clinical Immunology, 8th Edition, Daniel P. Sties, Abba I. Terr and Tristram G. Parsolw (eds), Appleton & Lange, Norwalk, CT, 1994. Light chains derived from any vertebrate species can be assigned to one of two distinct types called kappa and lambda based on the amino acid sequence of their constant domains. Immunoglobulins can be assigned to different classes or isotypes depending on the amino acid sequence of the constant domain (CH) of their heavy chain. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, each with a heavy chain denoted as α, δ, ε, γ, and μ, respectively. The γ and α classes are further divided into subclasses based on relatively minor differences in CH sequence and function. For example, humans express the following subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2.

[0146] As used herein, the terms “hypervariable region” or “HVR” refer to each region of an antibody variable domain whose sequence is hypervariable and / or which forms a structurally defined loop. Generally, antibodies contain six HVRs, three in VH (H1, H2, H3) and three in VL (L1, L2, L3). In native antibodies, H3 and L3 exhibit the highest diversity among the six HVRs, and H3 in particular is thought to play a unique role in conferring superior specificity to the antibody. See, for example, Xu et al., Immunity. 13 :37-45(2000), Johnson and Wu,in Methods in Molecular Biology 248 See 1-25 (Lo, ed., Human Press, Totowa, NJ, 2003). In fact, naturally occurring camel antibodies consisting only of heavy chains are functional and stable in the absence of light chains. For example, Hamers-Casterman et al., Nature. 363 :446-448(1993), Sheriff et al., Nature Struct.Biol. 3 See 733-736 (1996).

[0147] Several HVR descriptions are used and incorporated herein. Kabat complementarity-determining regions (CDRs), based on sequence variability, are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). Alternatively, Chothia refers to the location of structural loops (Chothia and Lesk, J. Mol. Biol.). 196:901-917(1987)). The AbM HVR represents a compromise between the Kabat HVR and the Chothia structural loop and is used by Oxford Molecular's AbM antibody modeling software. The "contact" HVR is based on the analysis of available complex crystal structures. The residues from each of these HVRs are listed in Table 1 below.

Table 1

[0148] The HVRs can include the following "extended HVRs". In VL, 24 - 36 or 24 - 34 (L1), 46 - 56 or 50 - 56 (L2), and 89 - 97 or 89 - 96 (L3), and in VH, 26 - 35 (H1), 50 - 65 or 49 - 65 (H2), and 93 - 102, 94 - 102, or 95 - 102 (H3). The variable domain residues are numbered according to the aforementioned Kabat et al. for each of these definitions.

[0149] The expressions "variable domain residue numbering as in Kabat" or "amino acid position numbering as in Kabat", and their variants, refer to the numbering system used in the heavy chain variable domain or the light chain variable domain in the antibody compilation in the aforementioned Kabat et al. Using this numbering system, the actual linear amino acid sequence can contain fewer amino acids or additional amino acids corresponding to deletions or insertions in the FR or HVR of the variable domain. For example, the heavy chain variable domain can include a single amino acid insertion (residue 52a according to Kabat) after residue 52 of H2, and residues inserted after residue 82 of the heavy chain FR (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). The Kabat numbering of residues can be determined for a given antibody by alignment in the homologous regions between the antibody's sequence and the sequence numbered by "standard" Kabat.

[0150] The term "variable" refers to the fact that specific segments of the variable domain vary widely in sequence among antibodies. The V domains mediate antigen binding and define the specificity of a particular antibody for its particular antigen. However, variability is not evenly distributed throughout the variable domain. Rather, it is concentrated in three segments called hypervariable regions (HVRs) in both the variable domains of the light and heavy chains. The more highly conserved portions of the variable domain are called framework regions (FRs). The native variable domains of the heavy and light chains each contain four FR regions that adopt a beta-sheet conformation that is largely connected by, and in some cases forms part of, the beta-sheet structure, and is connected by three HVRs that form loops that connect the beta-sheet structures. The HVRs within each chain are brought into proximity and linked to each other by the FR regions and, together with the HVRs of the other chain, contribute to the formation of the antigen-binding site of the antibody (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, MD (1991)). The constant domains do not directly participate in the binding of the antibody to antigen but exhibit various effector functions such as the participation of the antibody in antibody-dependent cytotoxicity.

[0151] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of the heavy or light chain of the antibody. The variable domains of the heavy and light chains may be referred to as "VH" and "VL", respectively. These domains are generally the most variable parts of the antibody (compared to other antibodies of the same class) and contain the antigen-binding site.

[0152] "Framework" or "FR" refers to the variable domain residues other than hypervariable region (HVR) residues. The FR of the variable domain generally consists of four FR domains: FR1, FR2, FR3 and FR4. Thus, the HVR and FR sequences generally appear in the following sequences in VH (or VL). FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4

[0153] The terms "full-length antibody," "intact antibody," and "whole antibody" are used interchangeably to refer to an antibody in its substantially intact form, as opposed to an antibody fragment. Specifically, a whole antibody includes antibodies with heavy and light chains containing an Fc region. The constant domain can be a natural sequence constant domain (e.g., human natural sequence constant domain) or an amino acid sequence variant thereof. In some cases, an intact antibody may have one or more effector functions.

[0154] An "antibody fragment" is a portion of an intact antibody, preferably comprising the antigen-binding region and / or variable region of the intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, diabodies, linear antibodies (see Example 2 of U.S. Patent No. 5,641,870, Zapata et al., Protein Eng. 8(10):1057-1062

[1995] ), single-chain antibody molecules, and multispecific antibodies formed from antibody fragments. Papain digestion of the antibody yields two identical antigen-binding fragments called "Fab" fragments and the remaining "Fc" fragment, a name reflecting its ability to readily crystallize. The Fab fragment consists of the entire light chain and the variable region domain (V) of the heavy chain. H ), and the first constant domain of one heavy chain (C H 1) consists of each Fab fragment, which is monovalent with respect to antigen binding, i.e., has a single antigen-binding site. Pepsin treatment of the antibody yields a single large F(ab')2 fragment, which is roughly equivalent to two Fab fragments with different antigen-binding activities disulfide-linked together, and is still capable of crosslinking to an antigen. The Fab' fragment contains one or more cysteine ​​derived from the hinge region of the antibody, C H It differs from Fab fragments in that it has several additional residues at the carboxyl terminus of one domain. Fab'-SH is the herein designation for Fab' in which one or more cysteine ​​residues in the constant domain have a free thiol group. The F(ab')2 antibody fragment was originally generated as a pair of Fab' fragments with a hinged cysteine ​​in between. Other chemical couplings of antibody fragments are also known.

[0155] The Fc fragment contains the carboxyl-terminal portions of both H chains held together by a disulfide. The effector function of the antibody is determined by the sequence within the Fc region, which is also the region recognized by the Fc receptor (FcR) found in specific cell types.

[0156] The “functional fragments” of the antibodies described include a portion of an intact antibody, which generally includes the antigen-binding region or variable region of an intact antibody, or the Fc region of an antibody that retains or modifies FcR-binding ability. Examples of antibody fragments include linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0157] "Fv" refers to the smallest antibody fragment containing a complete antigen recognition and binding site. This fragment consists of a dimer of one strongly non-covalently associated heavy chain variable domain and one light chain variable domain. The folding of these two domains generates six hypervariable loops (three from the H chain and three from the L chain) that contribute amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three antigen-specific HVRs) has the ability to recognize and bind to an antigen, albeit with lower affinity than the full binding site.

[0158] "Single-chain Fv," also abbreviated as "sFv" or "scFv," is a single polypeptide chain formed by linking together V molecules. H and V L It is an antibody fragment containing an antibody domain. Preferably, the sFv polypeptide is V H Domain and V LThe sFv further contains a polypeptide linker between the domain and the sFv, which allows the sFv to form a structure desirable for antigen binding. For an overview of sFv, see Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, edited by Rosenburg and Moore, Springer-Verlag, New York, pp. 269-315 (1994).

[0159] In this specification, the term “Fc region” is used to define the C-terminal region of an immunoglobulin heavy chain, and includes native sequence Fc regions and variant Fc regions. While the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region is typically defined as extending from the amino acid residue at position Cys226 or from Pro230 to its carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) can be removed, for example, during antibody production or purification, or by recombination of the nucleic acid encoding the antibody heavy chain. Therefore, intact antibody compositions may include antibody populations from which all K447 residues have been removed, antibody populations from which K447 residues have not been removed, and antibody populations containing mixtures of antibodies with and without K447 residues. Suitable native sequence Fc regions for use in the antibodies described include human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4. Unless otherwise specified herein, the numbering of amino acid residues within the Fc region or constant region follows the EU numbering, also known as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0160] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. The preferred FcR is the native human FcR sequence. Furthermore, preferred FcRs are those that bind to IgG antibodies (gamma receptors), including the FcγRI, FcγRII, and FcγRIII subclass receptors (including allele variants and, alternatively, splice forms of these receptors). The FcγRII receptor includes FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), which have similar amino acid sequences, differing primarily in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activating motif (ITAM) in its cytoplasmic domain. The inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic domain. (See M. Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs have been reviewed in Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991), Capel et al., Immunomethods 4:25-34 (1994), and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein.

[0161] The term "diabody" refers to the process of achieving inter-chain V-domain pairing, rather than intra-chain pairing, thereby obtaining a bivalent fragment, i.e., a fragment having two antigen-binding sites. H Domain and V L This refers to a small antibody fragment prepared by constructing an sFv fragment (see previous paragraph) using a short linker (approximately 5-10 residues) between the domain and the sFv. A bispecificity diabody is a V of two antibodies. H Domain and V LIt is a heterodimer of two “crossed” sFv fragments whose domains are located on polypeptide chains with different domains. Diabodies are described in detail, for example, European Patent No. 404,097, International Publication No. 93 / 11161, and Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993).

[0162] In this specification, monoclonal antibodies include, specifically, “chimeric” antibodies (immunoglobulins) in which a portion of the heavy chain and / or light chain is identical or homologous to a corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, while the rest of the chain is identical or homologous to a corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, insofar as they exhibit the desired biological activity (U.S. Patent No. 4,816,567, Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). In this specification, the chimeric antibody of interest includes PRIMATIZED® antibody, in which the antigen-binding region of the antibody is derived, for example, from an antibody produced by immunizing a macaque monkey with the antigen of interest. As used herein, “humanized antibody” is used as a subset of “chimeric antibody.”

[0163] The "class" of an antibody refers to the type of constant domain or constant region held by its heavy chain. There are five main classes of antibodies: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0164] "Affinity" refers to the strength of the total non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen, e.g., CD20). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects the 1:1 interaction between the members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y is generally expressed by the dissociation constant (K). D ) can be expressed by the following. Affinity can be measured by methods common in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0165] A “human antibody” is an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human, and / or an antibody prepared using any of the techniques for preparing human antibodies disclosed herein. This definition of a human antibody explicitly excludes humanized antibodies containing non-human antigen-binding residues. Human antibodies can be prepared using a variety of techniques known in the art, including phage display libraries. (Hoogenboom and Winter, J.Mol.Biol., 227:381 (1991), Marks et al., J.Mol.Biol., 222:581 (1991)). Furthermore, methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p.77 (1985), and Boerner et al., J.Immunol., 147(1):86-95 (1991) are available for the preparation of human monoclonal antibodies. van Dijk and van de Winkel,Curr.Opin.Pharmacol., 5See also 368-74 (2001). Human antibodies can be prepared by administering antigens to transgenic animals, such as immunized xenomouses, which have been modified to produce such antibodies in response to antigen administration, but whose endogenous gene loci are rendered inactive (see, for example, U.S. Patents 6,075,181 and 6,150,584 relating to XENOMOUSE® technology). For human antibodies produced by human B-cell hybridoma technology, see, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006).

[0166] Humanized non-human (e.g., mouse) antibodies are chimeric antibodies containing the smallest sequence derived from non-human immunoglobulin. In one embodiment, the humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's HVR (defined below) are replaced with residues from the HVR of a non-human species (donor antibody), such as mouse, rat, rabbit, or non-human primate, having desired specificity, affinity, and / or capabilities. In some cases, framework ("FR") residues of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, the humanized antibody may contain residues not found in either the recipient antibody or the donor antibody. These modifications may be made to further improve the antibody's performance, such as binding affinity. Generally, a humanized antibody includes at least one, typically two, variable domains substantially all of which, all or substantially all of the hypervariable loops, correspond to a non-human immunoglobulin sequence, and all or substantially all of the FR region corresponds to a human immunoglobulin sequence, although the FR region may contain one or more individual FR residue substitutions that improve antibody performance such as binding affinity, isomerization, and immunogenicity. The number of these amino acid substitutions in the FR is typically 6 or less in the H chain and 3 or less in the L chain. The humanized antibody also optionally includes an immunoglobulin constant region (Fc), typically at least a portion of the human immunoglobulin. For further details, see, for example, Jones et al., Nature 321:522-525 (1986), Riechmann et al., Nature 332:323-329 (1988), and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). For example, see Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1:105-115 (1998), Harris, Biochem. Soc. Transactions 23:1035-1038 (1995), Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994), and U.S. Patent Nos. 6,982,321 and 7,087,409.

[0167] The term “isolated antibody,” when used to describe the various antibodies disclosed herein, means an antibody identified, isolated, and / or recovered from cells or cell cultures on which the antibody is expressed. Contaminations from its natural environment are typically materials that interfere with the diagnostic or therapeutic use of the polypeptide and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody is purified to a purity of over 95% or over 99%, as determined, for example, by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse-phase HPLC). For a review of methods for evaluating antibody purity, see, for example, Flatman et al., J.Chromatogr.B 848:79-87 (2007). In preferred embodiments, the antibody is purified (1) using a spinning cup sequencer to a degree sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence, or (2) using Coomassie blue or preferably silver staining, by SDS-PAGE under non-reducing or reducing conditions until homogeneous. Isolated antibodies include in situ antibodies from recombinant cells, because at least one component of the polypeptide's natural environment is absent. However, the isolated polypeptide is usually prepared by at least one purification step.

[0168] As used herein, the term “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous antibody population; that is, the individual antibodies in that population are identical except for any naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation) that may be present in trace amounts. Monoclonal antibodies are highly specific and target a single antigenic site. In contrast to polyclonal antibody preparations, which typically contain different antibodies targeting different determinants (epitopes), each monoclonal antibody targets a single determinant on an antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they are synthesized by hybridoma culture and are not contaminated with other immunoglobulins. The modifier “monoclonal” indicates a characteristic of the antibody that it is obtained from a substantially homogeneous antibody population and should not be interpreted as requiring antibody production by any particular method. For example, monoclonal antibodies used in accordance with the present invention include, for example, hybridoma methods (e.g., Kohler and Milstein, Nature, 256:495-97 (1975), Hongo et al., Hybridoma, 14(3):253-260 (1995), Harlow et al., Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 2nd ed. 1988), Hammerling et al., in: Monoclonal Antibodies and T Cell Hybridomas 563-681 (Elsevier, New York (1981)), recombinant DNA methods (see, for example, U.S. Patent No. 4,816,567), phage display technology (e.g., Clackson et al., Nature, 352:624-628 (1991), Marks et al.) al., J.Mol.Biol.222:581-597(1992), Sidhu et al., J.Mol.Biol.338(2):299-310(2004), Lee et al. al., J.Mol.Biol.340(5):1073-1093(2004), Fellouse,Proc.Natl.Acad.Sci.USA 101(34):12467-12472(2004), and Lee et al.See J.Immunol.Methods 284(1-2):119-132(2004), as well as techniques for producing human antibodies or human-like antibodies in animals having some or all of the human immunoglobulin locus or genes encoding human immunoglobulin sequences (e.g., International Publication No. 1998 / 24893, International Publication No. 1996 / 34096, International Publication No. 1996 / 33735, International Publication No. 1991 / 10741, Jakobovits et al., Proc.Natl.Acad.Sci.USA 90:2551(1993), Jakobovits et al., Nature 362:255-258(1993), Bruggemann et al., Year in Immunol. 7:33 (1993), U.S. Patent Nos. 5,545,807, 5,545,806, 5,569,825, 5,625,126, 5,633,425, and 5,661,016, Marks et al., Bio / Technology 10:779-783 (1992), Lonberg et al., Nature 368:856-859 (1994), Morrison, Nature 368:812-813 (1994), Fishwild et al., Nature Biotechnol. 14:845-851 (1996), Neuberger, Nature Biotechnol. 14:826 (1996), and Lonberg et al. It can be prepared using various techniques, such as those described by Huszar, Intern. Rev. Immunol. 13:65-93 (1995).

[0169] As used herein, the terms “to bind,” “specifically bind to,” or “specific to” refer to measurable and reproducible interactions, such as binding, between a target and an antibody, which determine the presence of the target in the presence of a heterogeneous population of molecules, including biomolecules. For example, an antibody that specifically binds to a target (which may be an epitope) is an antibody that binds to this target more readily and / or for a longer period of time with higher affinity and binding activity than it would to other targets. In one embodiment, the extent to which the antibody binds to unrelated targets is less than about 10% of the antibody binding to the target, as measured, for example, by radioimmunoassay (RIA). In certain embodiments, an antibody that specifically binds to a target has a dissociation constant (K) of 1 μM or less, 100 nM or less, 10 nM or less, 1 nM or less, or 0.1 nM or less. D ) has. In certain embodiments, the antibody specifically binds to an epitope on a protein that is conserved between different species of proteins. In other embodiments, specific binding may include, but is not required, exclusive binding. The terms used herein are, for example, 10 -4 M or less, or 10 -5 M or less, or 10 -6 M or less, or 10 -7 M or less, or 10 -8 M or less, or 10 -9 M or less, or 10 -10 M or less, or 10 -11 M or less, or 10 -12 K against targets below M D , or 10 -4 M~10 -6 M or 10 -6 M~10 -10 M or 10 -7 M~10 -9 K in the range of M D This can be represented by molecules having affinity and K. As will be understood by those skilled in the art, affinity and K D The values ​​are inversely correlated. High affinity for the antigen is associated with low K DIt is measured by a value. In one embodiment, the term "specific binding" refers to a binding in which a molecule binds to a specific polypeptide or an epitope on a specific polypeptide without substantially binding to any other polypeptide or polypeptide epitope.

[0170] As used herein, the expressions "substantially reduced" or "substantially different" represent a sufficiently high difference between two numerical values (generally, one is associated with a certain molecule and the other is associated with a reference / comparative molecule), such that as a result, a person skilled in the art would consider the difference between these two values to be statistically significant with respect to the biological characteristic measured by such values (e.g., K D value). The difference between the two values is, depending on the value of the reference / comparative molecule, for example, more than about 10%, more than about 20%, more than about 30%, more than about 40%, and / or more than about 50%.

[0171] As used herein, the terms "substantially similar" or "substantially the same" indicate a sufficiently high similarity between two numerical values (e.g., one is associated with an antibody of the present invention and the other is associated with a reference / comparative antibody), such that as a result, a person skilled in the art would consider the difference between these two values to have little or no biological and / or statistical significance with respect to the biological characteristic measured by such values (e.g., K D value). The difference between the two values is, depending on the reference / comparative value, for example, less than about 50%, less than about 40%, less than about 30%, less than about 20%, and / or less than about 10%.

[0172] The "amino acid sequence identity percentage (%)" for a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence, after sequence alignment and, if necessary, introducing gaps to achieve the maximum possible sequence identity percentage, with no conservative substitutions considered as part of the sequence identity. Alignment for the purpose of determining the amino acid sequence identity percentage can be achieved in various ways within the skill of the art using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. A person skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm necessary to achieve the maximum possible alignment over the entire length of the sequences to be compared. However, for the purposes of this specification, the amino acid sequence identity % value is generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was created by Genentech, Inc., and its source code, along with user documentation, was filed with the U.S. Copyright Office, Washington DC, 20559, where it is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California, or can be compiled from source code. The ALIGN-2 program needs to be compiled for use with UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and remain unchanged.

[0173] In situations where ALIGN-2 is used for amino acid sequence comparison, the amino acid sequence identity % between a given amino acid sequence A and a given amino acid sequence B, between a given amino acid sequence A and a given amino acid sequence B, or between a given amino acid sequence A and a given amino acid sequence B (or, to put it another way, a given amino acid sequence A that has or contains a certain amino acid sequence identity % between a given amino acid sequence B, between a given amino acid sequence B and a given amino acid sequence B, or with respect to a given amino acid sequence B) is calculated as follows: 100 x fraction X / Y

[0174] (In the formula, X is the number of amino acid residues scored as identical matches in the alignment of A and B by the sequence alignment program ALIGN-2, and Y is the total number of amino acid residues in B.) It will be understood that if the length of amino acid sequence A is not equal to the length of amino acid sequence B, the amino acid sequence identity % of A vs. B will not be equal to the amino acid sequence identity % of B vs. A. Unless otherwise specified, all amino acid sequence identity % values ​​used herein are obtained using the ALIGN-2 computer program as described in the preceding paragraph.

[0175] As used herein, the term “sample” refers to a composition obtained from or derived from the subject and / or individual of interest, containing cellular and / or other molecular entities characterized and / or identified, for example, based on physical, biochemical, chemical, and / or physiological properties. For example, the terms “tumor sample,” “disease sample,” and variations thereof refer to any sample obtained from the subject of interest (e.g., a biopsy sample or a blood sample) that is expected to contain or is known to contain the characterized cellular and / or molecular entities. In some embodiments, the sample is a tumor tissue sample (e.g., a lymphoma tumor tissue sample, e.g., a B-cell lymphoma tumor tissue sample, e.g., a non-Hodgkin lymphoma tumor tissue sample, e.g., a diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma) tumor tissue sample). Other samples include, but are not limited to, primary or cultured cells or cell lines, cell supernatants, cell lysates, platelets, serum, plasma, vitreous fluid, lymph, synovial fluid, follicular fluid, semen, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, feces, tumor lysates, and tissue culture media, tissue extracts, such as homogenized tissue, cell extracts, and combinations thereof. Samples may be fresh or processed for preservation (e.g., frozen, fixed, or formalin-fixed, paraffin-embedded (FFPE)).

[0176] "Tissue sample" or "cell sample" means a collection of similar cells obtained from the tissue of a subject or individual. The source of a tissue or cell sample may be solid tissue such as fresh, frozen, and / or preserved organs, tissue samples, biopsy samples, and / or aspirates; blood such as plasma or any blood components; body fluids such as cerebrospinal fluid, amniotic fluid, ascites, or interstitial fluid; or cells at any stage in the subject's pregnancy or development. Tissue samples may be primary or cultured cells or cell lines. Optionally, tissue or cell samples may be obtained from diseased tissue / organs. Tissue samples may contain compounds that do not naturally mix with natural tissues, such as preservatives, anticoagulants, buffers, fixatives, waxes, nutrients, or antibiotics.

[0177] As used herein, “reference sample,” “reference cell,” “reference tissue,” “control sample,” “control cell,” or “control tissue” refers to a sample, cell, tissue, standard, or level used for comparative purposes. In one embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and / or non-disease portion (e.g., tissue or cell) of the same body of the subject. For example, healthy and / or non-disease cells or tissue adjacent to diseased cells or tissue (e.g., cells or tissue adjacent to a tumor). In another embodiment, the reference sample is obtained from untreated tissue and / or cells of the same body of the subject. In yet another embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and / or non-disease portion (e.g., tissue or cell) of a body of a non-subject. In yet another embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from untreated tissue and / or cells of a body of a non-subject individual.

[0178] Generally, when used in the context of the present invention, non-limiting examples of “control” are preferably “non-responder” controls, for example, samples / cells / tissues obtained from one or more patients who do not have a specific lymphoma as defined herein (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) (and are not “patients as defined herein”) and are known not to respond favorably to the anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and anti-CD20 antibody (e.g., obinutuzumab or rituximab) according to the present invention (especially in combination with chemotherapy, more specifically in combination with CHP chemotherapy). Another example of a “non-responder” control is a cell line / sample / cell / tissue that does not show an improved response to an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) in ex vivo studies (especially in combination with chemotherapy, especially in combination with CHP chemotherapy). Another non-limiting example of a “control” is an “internal standard,” e.g., a purified or synthetically produced protein, peptide, DNA and / or RNA, or a mixture thereof, the amount of each protein / peptide / DNA / RNA measured using the “non-responder” control described herein. In principle, the patient treated in the context of the present invention is assumed to be a patient with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). In other words, the patient is a patient who has / is suffering from lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)).Therefore, it is particularly assumed that the patients defined in relation to any aspect / embodiment are also patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)), and patients who have / are suffering from lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). However, it is not necessarily required that a given patient be diagnosed with lymphoma before (or after) the determination / identification / diagnosis that, for example, the patient is a patient defined in one or more aspects / embodiments as defined herein. However, it is preferable that the patient treated according to the present invention is diagnosed as a lymphoma patient in the first step and determined / identified / diagnosed as a patient as defined herein, particularly as defined in one or more aspects / embodiments, in the second step. In principle, according to the present invention, a given patient may be determined / identified / diagnosed as a patient as defined herein in the first step and diagnosed as a lymphoma patient in the second step. However, the latter option is less preferable, and as described above, the (previous or subsequent) step of diagnosing whether the patient to be treated is a lymphoma patient (e.g., DLBCL) can also be omitted.

[0179] Unless otherwise specified, the term “protein” as used herein refers to any native protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice, rats), unless otherwise specified. This term also includes any form of protein resulting from “full length,” untreated protein, and intracellular processing. This term also encompasses naturally occurring variants of proteins, such as splice variants or allele variants.

[0180] As used herein, “polynucleotide” or “nucleic acid” refers to a polymer of nucleotides of any length, including DNA and RNA. The nucleotides may be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or analogs thereof, or any substrate that can be incorporated into the polymer by DNA or RNA polymerase or by synthetic reactions. Therefore, examples of polynucleotides as defined herein include, but are not limited to, single-stranded and double-stranded DNA, DNA containing single-stranded and double-stranded regions, single-stranded and double-stranded RNA, RNA containing single-stranded and double-stranded regions, and hybrid molecules containing single-stranded or more typically double-stranded DNA and RNA, or potentially containing single-stranded and double-stranded regions. In addition, as used herein, the term “polynucleotide” refers to a triple-stranded region containing RNA or DNA, or both RNA and DNA. The strands within such a region may originate from the same molecule or from different molecules. These regions may contain all of one or more of these molecules, but more typically, only some of these molecules. One of the molecules in a triple helix region is often an oligonucleotide. The terms "polynucleotide" and "nucleic acid" specifically include mRNA and cDNA.

[0181] Polynucleotides may include modified nucleotides such as methylated nucleotides and their analogues. If present, modifications to the nucleotide structure may be applied before or after polymer assembly. The nucleotide sequence may be interrupted by non-nucleotide components. Polynucleotides may be further modified after synthesis, such as by conjugation with labels. Other types of modifications include, for example, "caps," substitution of one or more naturally occurring nucleotides with analogs, internucleotide modifications, such as those by uncharged bonds (e.g., methylphosphonates, phosphotriesters, phosphoamidates, carbamates, etc.) and those by charged bonds (e.g., phosphorothioates, phosphorodithioates, etc.), suspensions, such as those containing proteins (e.g., nucleases, toxins, antibodies, signal peptides, poly-L-lysine, etc.), those by intercalating agents (e.g., acridine, psoralens, etc.), those containing chelating agents (e.g., metals, radioactive metals, boron, metal oxides, etc.), those containing alkylating agents, those by modified bonds (e.g., alpha-anomeric nucleic acids, etc.), and the unmodified form of polynucleotides. Furthermore, any of the hydroxyl groups normally present in the sugar may be replaced with, for example, a phosphonic acid group or a phosphate group, protected with a standard protecting group, or activated to prepare for additional binding to additional nucleotides, or conjugated to a solid or semi-solid support. The 5' and 3' terminal OH groups may be phosphorylated or substituted with amines or organic capping groups of 1 to 20 carbon atoms. Other hydroxyls may also be derivatized with standard protecting groups. Polynucleotides may also include analogous forms of ribose or deoxyribose sugars commonly known in the art, such as 2'-O-methyl-, 2'-O-allyl-, 2'-fluoro-, or 2'-azid-ribose, carbocyclic sugar analogs, α-anomeric sugars, epimeric sugars, such as arabinose, xylose, or lyxose, pyranose sugars, furanose sugars, sedoheptulose, acyclic analogs, and nonbasic nucleoside analogs, such as methylriboside.One or more phosphodiester bonds may be replaced by alternative linking groups. Examples of these alternative linking groups include, but are not limited to, embodiments in which the phosphate is replaced by P(O)S ("thioate"), P(S)S ("dithioate"), (O)NR2 ("amidate"), P(O)R, P(O)OR', CO, or CH2 ("formacetal"), where each R or R' is independently H, or a substituted or unsubstituted alkyl (1-20C) (optionally including an ether (-O-) bond), aryl, alkenyl, cycloalkyl, cycloalkenyl, or aralgyl. Not all bonds in a polynucleotide need to be identical. The foregoing description applies to all polynucleotides referred to herein, including RNA and DNA.

[0182] As used herein, “carrier” includes pharmaceutically acceptable carriers, excipients, or stabilizers that are nontoxic to cells or mammals to which they are exposed at the dosage and concentration used. In many cases, physiologically acceptable carriers are pH-buffered aqueous solutions. Examples of physiologically acceptable carriers include buffers such as phosphoric acid, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN®, polyethylene glycol (PEG), and PLURONICS®.

[0183] The phrase "pharmaceutically acceptable" indicates that a substance or composition must be chemically and / or toxicologically compatible with the other components of the formulation and / or the mammal being treated with it.

[0184] The term "pharmaceutical preparation" refers to a preparation in which the biological activity of the active ingredient contained therein is effective, and which does not contain any further components that are unacceptably toxic to the person to whom the preparation is administered.

[0185] "Product" is any product (e.g., package or container) or kit comprising at least one reagent, for example, a pharmaceutical for treating a disease or disorder (e.g., lymphoma, e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)), and a package insert. In certain embodiments, the product or kit is promoted, distributed, or sold as a unit for carrying out the methods described herein.

[0186] "Package insert" refers to instructions that are typically included in the commercial packaging of a pharmaceutical product and contain information regarding indications, dosage, administration, contraindications, other pharmaceuticals to be used in combination with the packaged product, and / or warnings regarding the use of such pharmaceutical product.

[0187] As used herein, the term “CD79b” refers to any natural CD79b from any vertebrate source, including mammals such as primates (e.g., humans, cynomolgus monkeys (cynos)) and rodents (e.g., mice and rats), unless otherwise indicated. In this specification, human CD79b is also referred to as “Igβ,” “B29,” “DNA225786,” or “PRO36249.” An exemplary CD79b sequence including a signal sequence is shown in SEQ ID NO: 1. An exemplary CD79b sequence without a signal sequence is shown in SEQ ID NO: 2. The term “CD79b” encompasses “full-length” unprocessed CD79b, as well as any form of CD79b resulting from cellular processing. The term also encompasses naturally occurring variants of CD79b, such as splice variants, allele variants, and isoforms. The CD79b polypeptides described herein may be isolated from diverse sources, such as human tissue species or other sources, or prepared by recombinant or synthetic methods. “Natural sequence CD79b polypeptide” includes polypeptides having the same amino acid sequence as the corresponding naturally occurring CD79b polypeptide. Such natural sequence CD79b polypeptides may be isolated from nature or produced by recombinant or synthetic methods. The term “natural sequence CD79b polypeptide” specifically includes naturally occurring transcribed or secreted forms of a particular CD79b polypeptide (e.g., extracellular domain sequence), naturally occurring variant forms (e.g., alternative splicing forms), and naturally occurring allelic variants of the polypeptide.

[0188] The terms "anti-CD79b antibody" or "CD79b-binding antibody" refer to an antibody capable of binding to CD79b with sufficient affinity so that it is useful as a diagnostic and / or therapeutic agent in targeting CD79b. Preferably, the degree of binding of an anti-CD79b antibody to unrelated non-CD79b proteins is less than about 10% of the binding of the antibody to CD79b as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the dissociation constant (Kd value) of the antibody binding to CD79b is ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, or ≤0.1 nM. In certain embodiments, the anti-CD79b antibody binds to an epitope of CD79b that is conserved among different species of CD79b.

[0189] The term "CD79b-positive cancer" refers to cancers that include cells expressing CD79b on their surface. In some embodiments, CD79b expression on the cell surface is determined using antibodies against CD79b by methods such as immunohistochemistry or FACS. Alternatively, CD79b mRNA expression is thought to correlate with CD79b expression on the cell surface and can be determined by methods selected from in situ hybridization and RT-PCR (including quantitative RT-PCR).

[0190] "Alkyl" refers to a C1-C group containing normal, secondary, tertiary, or cyclic carbon atoms. 18 These are hydrocarbons. Examples include methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), and 2-methyl-2-propyl ( t -Bu, t -butyl, -C(CH3)3), 1-pentyl n-Pentyl (-CH2CH2CH2CH2CH3), 2-Pentyl (-CH(CH3)CH2CH2CH3), 3-Pentyl (-CH(CH2CH3)2), 2-Methyl-2-butyl (-C(CH3)2CH2CH3), 3-Methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-Methyl-1-butyl (-CH2CH2CH(CH3)2), 2-Methyl-1-butyl (-CH2CH(CH3)CH2CH3), 1-Hexyl (-CH2CH2CH2CH2CH2CH3), 2-Hexyl (-CH(CH3)CH2CH2CH2CH3), 3-Hexyl (-C These are H(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl(-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl(-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl(-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl(-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl(-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl(-C(CH3)2CH(CH3)2), and 3,3-dimethyl-2-butyl(-CH(CH3)C(CH3)3).

[0191] As used herein, the term "C1-C8 alkyl" refers to a linear or branched saturated or unsaturated hydrocarbon having 1 to 8 carbon atoms. Representative "C1-C8 alkyl" groups include, but are not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl, -n-octyl, -n-nonyl, and -n-decyl, while branched C1-C8 alkyl groups include, but are not limited to, -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, and 2-methylbutyl, and unsaturated C1-C8 alkyl groups. Examples of rukyls, though not limited to them, include -vinyl, -allyl, -1-butenyl, -2-butenyl, -isobutyrenyl, -1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-dimethyl-2-butenyl, -1-hexyl, -2-hexyl, -3-hexyl, -acetylenyl, -propynyl, -1-butynyl, -2-butynyl, -1-pentynyl, -2-pentynyl, and -3-methyl-1-butynyl. The C1-C8 alkyl group may be unsubstituted or substituted with one or more groups including, but not limited to, -C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2-NHC(O)R', -SO3R', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN, where each R' is independently selected from H, -C1-C8 alkyl, and aryl.

[0192] When used herein, "C1~C 12 The term "alkyl" refers to a linear or branched saturated or unsaturated hydrocarbon having 1 to 12 carbon atoms. 12The alkyl group may be unsubstituted or substituted with one or more groups including, but not limited to, -C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2-NHC(O)R', -SO3R', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN, where each R' is independently selected from H, -C1-C8 alkyl, and aryl.

[0193] As used herein, the term "C1-C6 alkyl" refers to a linear or branched saturated or unsaturated hydrocarbon having 1 to 6 carbon atoms. Representative "C1-C6 alkyl" groups include, but are not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl, while branched C1-C6 alkyls include, but are not limited to, -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, and 2-methylbutyl, and unsaturated C1-C6 alkyls include, but are not limited to, -vinyl, -allyl, -1-butenyl, -2-butenyl, and -isobutyrenyl, -1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-dimethyl-2-butenyl, 1-hexyl, 2-hexyl, and 3-hexyl. The C1-C6 alkyl groups may be unsubstituted, or they may be substituted with one or more groups, as described above for the C1-C8 alkyl groups.

[0194] As used herein, the term "C1-C4 alkyl" refers to a linear or branched saturated or unsaturated hydrocarbon having 1 to 4 carbon atoms. Representative "C1-C4 alkyl" groups include, but are not limited to, -methyl, -ethyl, -n-propyl, and -n-butyl, while branched C1-C4 alkyls include, but are not limited to, -isopropyl, -sec-butyl, -isobutyl, and -tert-butyl, and unsaturated C1-C4 alkyls include, but are not limited to, -vinyl, -allyl, -1-butenyl, -2-butenyl, and -isobutylenyl. C1-C4 alkyl groups may be unsubstituted or substituted with one or more groups, as described above for C1-C8 alkyl groups.

[0195] An "alkoxy" is an alkyl group bonded to an oxygen atom alone. Examples of alkoxy groups include, but are not limited to, methoxy (-OCH3) and ethoxy (-OCH2CH3). A "C1-C5 alkoxy" is an alkoxy group having 1 to 5 carbon atoms. As with alkyl groups described above, alkoxy groups may be unsubstituted or substituted with one or more groups.

[0196] "Alkenyl" has at least one unsaturated site, i.e., carbon-carbon sp 2 These are C2-C18 hydrocarbons containing normal, secondary, tertiary, or cyclic carbon atoms with double bonds. Examples include, but are not limited to, ethylene or vinyl (-CH=CH2), allyl (-CH2CH=CH2), cyclopentenyl (-C5H7), and 5-hexenyl (-CH2CH2CH2CH2CH=CH2). "C2-C8 alkenyls" have at least one unsaturated site, i.e., carbon-carbon, sp 2 It is a hydrocarbon containing 2 to 8 normal, secondary, tertiary, or cyclic carbon atoms with double bonds.

[0197] "Alkynnyl" refers to a C2-C18 hydrocarbon containing at least one unsaturated site, i.e., a normal, secondary, tertiary, or cyclic carbon atom having a carbon-carbon sp triple bond. Examples, though not limited to them, include acetylene (-C≡CH) and propargyl (-CH2C≡CH). "C2-C8 alkynyl" refers to a hydrocarbon containing at least one unsaturated site, i.e., 2 to 8 normal, secondary, tertiary, or cyclic carbon atoms having a carbon-carbon sp triple bond.

[0198] "Alkylene" refers to a saturated, branched, linear, or cyclic hydrocarbon radical with 1 to 18 carbon atoms, having two monovalent radical centers derived by removing two hydrogen atoms from the same or two different carbon atoms of the parent alkane. Typical alkylene radicals, though not limited to these, include methylene(-CH2-)1,2-ethyl(-CH2CH2-), 1,3-propyl(-CH2CH2CH2-), and 1,4-butyl(-CH2CH2CH2CH2-).

[0199] "C1~C 10 Alkylene is represented by the formula (CH2) 1~10 - is a straight-chain saturated hydrocarbon group. C1~C 10 Examples of alkylenes include methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylene, and decalene.

[0200] "Alkenylene" refers to an unsaturated, branched, linear, or cyclic hydrocarbon radical with 2 to 18 carbon atoms, having two monovalent radical centers derived by removing two hydrogen atoms from the same or two different carbon atoms of the parent alkene. Typical alkenylene radicals include, but are not limited to, 1,2-ethylene (-CH=CH-).

[0201] "Alkynylene" refers to an unsaturated, branched, linear, or cyclic hydrocarbon radical with 2 to 18 carbon atoms, having two monovalent radical centers derived by removing two hydrogen atoms from the same or two different carbon atoms of the parent alkyne. Typical alkynylene radicals, but not limited to, include acetylene (-C≡C-), propagyl (-CH2C≡C-), and 4-pentinyl (-CH2CH2CH2C≡C-).

[0202] "Aryl" refers to a carbocyclic aromatic group. Examples of aryl groups, though not limited to them, include phenyl, naphthyl, and anthracenyl. The carbocyclic or heterocyclic aromatic group may be unsubstituted or substituted with one or more groups, including but not limited to -C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2-NHC(O)R', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN (wherein each R' is independently selected from H, -C1-C8 alkyl, and aryl).

[0203] "C5~C 20 "Aryl" refers to an aryl group having 5 to 20 carbon atoms in a carbocyclic aromatic ring. C5~C 20 Examples of aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl. C5~C 20 The aryl group can be substituted or unsubstituted, as described above. 14 "Aryl" refers to an aryl group having 5 to 14 carbon atoms in a carbocyclic aromatic ring. 14 Examples of aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl. C5~C 14 The aryl group can be substituted or unsubstituted, as described above.

[0204] "Arirene" is an aryl group having two covalent bonds and which can be in an ortho, meta, or para configuration as shown in the following structure: [ka] (In the formula, the phenyl group is either unsubstituted or substituted with up to four groups, including but not limited to -C1-C8 alkyl, -O-(C1~C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2-NHC(O)R', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2 and -CN, where each R' is independently selected from H, -C1~C8 alkyl and aryl).

[0205] "Arylalkyl" refers to carbon atoms, typically terminal or sp 3 This refers to an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom is replaced by an aryl radical. Typical arylalkyl groups, though not limited to these, include benzyl, 2-phenylethane-1-yl, 2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethane-1-yl, 2-naphthylethen-1-yl, naphthobenzyl, and 2-naphthophenylethane-1-yl. Arylalkyl groups contain 6 to 20 carbon atoms; for example, the alkyl portion of an arylalkyl group containing an alkanyl, alkenyl, or alkynyl group has 1 to 6 carbon atoms, while the aryl portion has 5 to 14 carbon atoms.

[0206] "Heteroarylalkyl" refers to carbon atoms, typically terminal or sp 3Refers to an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom is replaced by a heteroaryl radical. Typical heteroarylalkyl groups include, but are not limited to, 2-benzimidazolylmethyl, 2-furylethyl, etc. The heteroarylalkyl group contains 6 to 20 carbon atoms. For example, in the heteroarylalkyl group, the alkyl moiety such as an alkanoyl, alkenyl or alkynyl group has 1 to 6 carbon atoms, and the heteroaryl moiety has 5 to 14 carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S. The heteroaryl moiety of the heteroarylalkyl group has a monocyclic ring having 3 to 7 ring members (2 to 6 carbon atoms), or a bicyclic ring having 7 to 10 ring members (4 to 9 carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S), for example, a bicyclo[4,5], [5,5], [5,6] or [6,6] system.

[0207] "Substituted alkyl", "substituted aryl", and "substituted arylalkyl" each mean an alkyl, aryl, and arylalkyl in which one or more hydrogen atoms are each independently replaced by a substituent. Typical substituents include, but are not limited to, -X, -R, -O - , -OR, -SR, -S - , -NR2, -NR3, =NR, -CX3, -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO2, =N2, -N3, NC(=O)R, -C(=O)R, -C(=O)NR2, -SO3 - , -SO3H, -S(=O)2R, -OS(=O)2OR, -S(=O)2NR, -S(=O)R, -OP(=O)(OR)2, -P(=O)(OR)2, -PO - 3, -PO3H2, -C(=O)R, -C(=O)X, -C(=S)R, -CO2R, -CO2 - , -C(=S)OR, -C(=O)SR, -C(=S)SR, -C(=O)NR2, -C(=S)NR2, -C(=NR)NR2, where each X is independently a halogen: F, Cl, Br, or I, and each R is independently -H, C2~C 18 alkyl, C6~C 20Aryl, C3-C 14 is a heterocyclic ring, a protecting group or a prodrug moiety. The alkylene, alkenylene and alkynylene groups described above may similarly be substituted.

[0208] "Heteroaryl" and "heterocyclic ring" refer to ring systems in which one or more ring atoms are heteroatoms, such as nitrogen, oxygen and sulfur. The heterocyclic radical contains 3 to 20 carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S. The heterocyclic ring has a monocyclic ring with 3 to 7 ring members (2 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S), or a bicyclic ring with 7 to 10 ring members (4 to 9 carbon atoms and 1 to 3 heteroatoms selected from N, O, P and S), such as: a bicyclo[4,5], [5,5], [5,6], or [6,6] system.

[0209] Exemplary heterocyclic rings are described, for example, in Paquette, Leo A., "Principles of Modern Heterocyclic Chemistry" (W.A. Benjamin, New York, 1968), especially Chapters 1, 3, 4, 6, 7 and 9, and in "The Chemistry of Heterocyclic Compounds, A series of Monographs" (John Wiley & Sons, New York, 1950 - present), especially Volumes 13, 14, 16, 19, and 28, and in J. Am. Chem. Soc. (1960) 82:5566.

[0210] Examples of heterocycles, though not limited to them, include pyridyl, dihydroipyridyl, tetrahydropyridyl (piperidyl), thiazolyl, tetrahydrothiophenyl, sulfur-oxidized tetrahydrothiophenyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl, indolyl, indrenyl, quinolinyl, isoquinolinyl, benzimidazolyl, piperidinyl, and 4-piperyl. Donyl, pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl, tetrahydrofuran, bis-tetrahydrofuran, tetrahydropyranyl, bis-tetrahydropyranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, octahydroisoquinolinyl, azosinyl, triazinyl, 6H-1,2,5-thiadiadinyl, 2H,6H-1,5,2-dithiadinyl, thienyl, thianthrenyl, pyranyl, isobenzoph Lanyl, Clomenyl, Xanthenyl, Phenoxathinyl, 2H-Pyrrolyl, Isothiazolyl, Isoxazolyl, Pyrazinyl, Pyridazinyl, Indolidinyl, Isoindolyl, 3H-Indolyl, 1H-Indazolyl, Purinyl, 4H-Quinolidinyl, Phthalazinyl, Naphthylidinyl, Quinoxalinyl, Quinazolinyl, Synnolinyl, Pteridinyl, 4aH-Carbazolyl, Carbazolyl, β-Carborinyl, Phenantridinyl, Acrylidinyl Examples include nyl, pyrimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, flazanyl, phenoxazinyl, isochromanil, chromanil, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolidinyl, piperazinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl, oxazolidinyl, benzotriazolyl, benzoisoxazolyl, oxyndryl, benzoxazolinyl, and isatinoyl.

[0211] As an example, and not an exhaustive list, carbon-bonded heterocycles are bonded at positions 2, 3, 4, 5, or 6 of pyridine, positions 3, 4, 5, or 6 of pyridazine, positions 2, 4, 5, or 6 of pyrimidine, positions 2, 3, 5, or 6 of pyrazine, positions 2, 3, 5, or 5 of furan, tetrahydrofuran, thiofuran, thiophene, pyrrole, or tetrahydropyrrole, positions 2, 3, 4, or 5 of oxazole, imidazole, or thiazole, positions 3, 4, or 5 of isoxazole, pyrazole, or isothiazole, positions 2 or 3 of aziridine, positions 2, 3, or 4 of azetidine, positions 2, 3, 4, 5, 6, 7, or 8 of quinoline, or positions 1, 3, 4, 5, 6, 7, or 8 of isoquinoline. More typically, carbon-bonded heterocycles include 2-pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl, 6-pyridyl, 3-pyridazinyl, 4-pyridazinyl, 5-pyridazinyl, 6-pyridazinyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl, 5-pyrazinyl, 6-pyrazinyl, 2-thiazolyl, 4-thiazolyl, or 5-thiazolyl.

[0212] As an example, though not an exhaustive list, nitrogen-bonded heterocycles are linked at position 1 of aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole, position 2 of isoindole or isoindoline, position 4 of morpholine, and position 9 of carbazole or β-carbolin. More typically, nitrogen-bonded heterocycles include 1-aziridyl, 1-azetezyl, 1-pyrrolyl, 1-imidazolyl, 1-pyrazolyl, and 1-piperidinyl.

[0213] A "C3-C8 heterocycle" refers to an aromatic or non-aromatic C3-C8 carbon ring in which 1 to 4 of the ring carbon atoms are independently replaced by heteroatoms from the group consisting of O, S, and N. Representative examples of C3-C8 heterocycles, though not limited to them, include benzofuranyl, benzothiophene, indolyl, benzopyrazolyl, coumalinyl, isoquinolinyl, pyrrolyl, thiophenyl, furanyl, thiazolyl, imidazolyl, pyrazolyl, triazolyl, quinolinyl, pyrimidinyl, pyridinyl, pyridonyl, pyrazinyl, pyridadinyl, isothiazolyl, isoxazolyl, and tetrazolyl. The C3-C8 heterocycle may be unsubstituted or substituted with up to seven groups, including but not limited to -C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2-NHC(O)R', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN, where each R' is independently selected from H, -C1-C8 alkyl, and aryl.

[0214] "C3-C8 heterocyclo" refers to the C3-C8 heterocyclic group defined above, in which one of the hydrogen atoms of the heterocyclic group is replaced by a single bond. C3-C8 heterocyclo may be unsubstituted or substituted with up to six groups, including but not limited to -C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2-NHC(O)R', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN, where each R' is independently selected from H, -C1-C8 alkyl, and aryl.

[0215] "C3~C 20 A "heterocycle" refers to an aromatic or non-aromatic C3-C8 carbon ring in which one to four of the ring's carbon atoms are independently substituted by heteroatoms from the group consisting of O, S, and N. 20The heterocycle may be unsubstituted or substituted with up to seven groups, including but not limited to -C1-C8alkyl, -O-(C1-C8alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2-NHC(O)R', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2 and -CN, where each R' is independently selected from H, -C1-C8alkyl and aryl.

[0216] "C3~C 20 A heterocyclo is defined as a C3-C group in which one of the hydrogen atoms of the heterocyclic group is replaced by a single bond. 20 It refers to a heterocyclic group.

[0217] A "carbocyclic ring" refers to a saturated or unsaturated ring having 3 to 7 carbon atoms as a monocyclic ring or 7 to 12 carbon atoms as a dicyclic ring. Monocyclic carbocyclic rings have 3 to 6 ring atoms, and more typically 5 or 6 ring atoms. Dicyclic carbocyclic rings have 7 to 12 ring atoms arranged, for example, as bicyclo[4,5], [5,5], [5,6], or [6,6] systems, or 9 or 10 ring atoms arranged as bicyclo[5,6] or [6,6] systems. Examples of monocyclic carbocyclic rings include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopenta-1-enyl, 1-cyclopenta-2-enyl, 1-cyclopenta-3-enyl, cyclohexyl, 1-cyclohexa-1-enyl, 1-cyclohexa-2-enyl, 1-cyclohexa-3-enyl, cycloheptyl, and cyclooctyl.

[0218] A "C3-C8 carbon ring" is a saturated or unsaturated non-aromatic carbon ring with 3, 4, 5, 6, 7, or 8 members. Representative C3-C8 carbon rings, though not limited to them, include -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl, -1,3-cyclohexadienyl, -1,4-cyclohexadienyl, -cycloheptyl, -1,3-cycloheptadienyl, -1,3,5-cycloheptatrielinyl, -cyclooctyl, and -cyclooctadienyl. The C3-C8 carbocyclic group may be unsubstituted or substituted with one or more groups including, but not limited to, -C1-C8 alkyl, -O-(C1-C8 alkyl), -aryl, -C(O)R', -OC(O)R', -C(O)OR', -C(O)NH2, -C(O)NHR', -C(O)N(R')2-NHC(O)R', -S(O)2R', -S(O)R', -OH, -halogen, -N3, -NH2, -NH(R'), -N(R')2, and -CN, where each R' is independently selected from H, -C1-C8 alkyl, and aryl.

[0219] "C3-C8 carbocyclo" refers to the C3-C8 carbocyclic group defined above, in which one of the hydrogen atoms of the carbocyclic group is replaced by a single bond.

[0220] A "linker" refers to a chemical moiety containing a covalent bond or a chain of atoms that covalently bonds the antibody to the drug portion. In various embodiments, the linker may be a divalent radical such as alkyldiyl, aryldiyl, or heteroaryldiyl, or -(CR2) n O(CR2) n - Examples include repeating units of alkyloxy (e.g., polyethyleneoxy, PEG, polymethyleneoxy) and alkylamino (e.g., polyethyleneamino, Jeffamine®), as well as diacid esters and amides such as succinic acid esters, succinic acid amides, diglycolic acid esters, malonic acid esters, and caproamides. In various embodiments, the linker may contain one or more amino acid residues such as valine, phenylalanine, lysine, and homolysine.

[0221] The term "chiral" refers to molecules that cannot be superimposed on their mirror image partners, while the term "achiral" refers to molecules that can be superimposed on those mirror image partners.

[0222] The term "stereoisomer" refers to a compound that has the same chemical structure but differs in the arrangement of atoms or groups in space.

[0223] A "diastereomer" refers to a stereoisomer that has two or more chiral centers, and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral characteristics, and reactivity. Mixtures of diastereomers can be separated under high-resolution analytical procedures such as electrophoresis and chromatography.

[0224] An "enantiomer" refers to two stereoisomers of a compound that are mirror images of each other and cannot be superimposed.

[0225] The stereochemical definitions and conventions used herein generally follow SP. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York, and Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds (1994), John Wiley & Sons, Inc., New York. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule around its chiral center. The prefixes d and l or (+) and (-) are used to indicate the rotation of plane-polarized light by the compound, with (-) or l meaning that the compound is levorotatory. Compounds with the prefix (+) or d are dextrorotatory. In a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Certain stereoisomers are also called enantiomers, and mixtures of such isomers are sometimes called enantiomer mixtures. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, and these can occur when neither stereoselectivity nor stereospecificity is present in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to equimolar mixtures of two enantiomer species that are not optically active.

[0226] A "leaving group" refers to a functional group that can be substituted by another functional group. Certain leaving groups are well known in the art, and examples, though not limited to them, include halides (e.g., chlorides, bromides, iodides), methanesulfonyl (mesyl), p-toluenesulfonyl (tosyl), trifluoromethylsulfonyl (triflate), and trifluoromethylsulfonates.

[0227] The term "protecting group" refers to a substituent commonly used to block or protect a particular functionality in a compound, while reacting with other functional groups on the compound. For example, an "amino protecting group" is a substituent that attaches to an amino group to block or protect the amino functionality of a compound. Suitable amino protecting groups include, but are not limited to, acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). For a general explanation of protecting groups and their use, see TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991 or later editions.

[0228] III. Diagnostic Methods and Assays Methods and assays are provided herein for identifying, diagnosing and / or predicting whether patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) may benefit from treatments comprising an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab). The methods and assays described herein are based on the finding that patients can be identified, diagnosed, and / or predicted to benefit from treatments including an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) by using the amount or level of macrophage biomarkers (e.g., gene expression values ​​(e.g., gene expression values ​​derived from any of the gene signature sets described herein (e.g., any of the exemplary gene signature sets in Table 2 or Table 3)) or macrophage quantity (e.g., M1 macrophages or tumor-associated macrophages)) in patient samples (e.g., tissue samples, e.g., tumor tissue samples, e.g., biopsy samples, etc.)). Any of the methods provided herein may include administering an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) to a patient.

[0229] A. Macrophage biomarkers In certain cases, the amounts or levels of macrophage biomarkers can be determined using the methods and assays provided herein. Various diagnostic methods based on the determination of macrophage biomarker amounts or levels are further described below.

[0230] In one embodiment, a method is provided herein for identifying, diagnosing, and / or predicting whether a patient having lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) can benefit from a treatment comprising anti-CD79b immunoconjugate and anti-CD20 antibody, comprising measuring macrophage biomarkers in a sample from the patient, wherein the amount or level of macrophage biomarkers in the sample is below the amount or level of reference macrophage biomarkers, thereby identifying, diagnosing, and / or predicting that the patient will benefit from a treatment comprising anti-CD79b immunoconjugate and anti-CD20 antibody. In some examples, the method comprises administering anti-CD79b immunoconjugate and anti-CD20 antibody.

[0231] In another embodiment, a method is provided herein for selecting a therapy for a patient having lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) comprising measuring macrophage biomarkers in a sample from the patient, wherein the amount or level of macrophage biomarkers in the sample is below the amount or level of reference macrophage biomarkers, thereby identifying the patient as one who would benefit from a treatment comprising anti-CD79b immunoconjugate and anti-CD20 antibody. In some examples, the method comprises administering anti-CD79b immunoconjugate and anti-CD20 antibody.

[0232] (i) Decrease in macrophage biomarkers The amount or level of macrophage biomarkers in a sample from a patient with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) being below the amount or level of reference macrophage biomarkers may identify, diagnose, and / or predict that the patient may benefit from treatment including anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

[0233] In some cases, the amount or level of macrophage biomarkers in a sample is approximately the lower 99th percentile (prevalence level of approximately 99% or less), approximately the lower 95th percentile (prevalence level of approximately 95% or less), approximately the lower 90th percentile (prevalence level of approximately 90% or less), approximately the lower 85th percentile (prevalence level of approximately 85% or less), and approximately the lower 80th percentile compared to the amount or level of macrophage biomarkers in the reference population. The lower percentile (prevalence level of approximately 80% or less), the lower 75th percentile (prevalence level of approximately 75% or less), the lower 70th percentile (prevalence level of approximately 70% or less), the lower 65th percentile (prevalence level of approximately 65% ​​or less), the lower 60th percentile (prevalence level of approximately 60% or less), the lower 55th percentile (prevalence level of approximately 55% or less), the lower 50th percentile (prevalence level of approximately 50% or less), The lowest 45th percentile (prevalence level of approximately 45% or less), the lowest 40th percentile (prevalence level of approximately 40% or less), the lowest 35th percentile (prevalence level of approximately 35% or less), the lowest 30th percentile (prevalence level of approximately 30% or less), the lowest 25th percentile (prevalence level of approximately 25% or less), the lowest 20th percentile (prevalence level of approximately 20% or less), and the lowest 15th percentile (approximately 1 Individuals are identified as likely to benefit from treatments including anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) by being within the following prevalence levels: less than 5%, approximately the lower 10th percentile (less than 10%), approximately the lower 5th percentile (less than 5%), or approximately the lower 1st percentile (less than 1%).

[0234] In some cases, the amount or level of macrophage biomarkers in a sample being within the approximately lower 10th to 90th percentile, 20th to 80th percentile, 30th to 70th percentile, 40th to 60th percentile, 45th to 55th percentile, 48th to 52nd percentile, 49.5th to 50.5th percentile, 49.9th to 50.1st percentile, or approximately 50th percentile of macrophage biomarkers in the reference population identifies an individual as likely to benefit from treatments including anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab). For example, in some cases, the amount or level of macrophage biomarkers in a sample corresponds to a prevalence of approximately 10% to 90% in the reference population, approximately 15% to 85%, approximately 20% to 80%, approximately 25% to 75%, approximately 30% to 70%, approximately 35% to 65%, approximately 40% to 60%, and approximately 45% to 55%. Individuals are identified as likely to benefit from treatments including anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) if they have a prevalence of approximately 48% to 52%, 49.5% to 50.5%, 49.9% to 50.1%, or 50%.

[0235] In some cases, a macrophage biomarker amount or level lower than the reference amount or level refers to a decrease of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% or more compared to the amount or level of macrophage biomarker detected by standard methods known in the art, such as those described herein, compared to the amount or level of macrophage biomarker in a reference sample, reference cells, reference tissue, control sample, control cells, or control tissue. In certain cases, a macrophage biomarker amount or level lower than the reference amount or level refers to a decrease in the amount or level of macrophage biomarker in a sample, where the decrease is at least about 1.5 times, 1.75 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 25 times, 50 times, 75 times, or 100 times the amount or level of macrophage biomarker in the reference sample, reference cells, reference tissue, control sample, control cells, or control tissue. In some cases, a macrophage biomarker amount or level lower than the reference amount or level refers to a decrease in the amount or level of macrophage biomarker that is greater than about 1.5 times, about 1.75 times, about 2 times, about 2.25 times, about 2.5 times, about 2.75 times, about 3.0 times, or about 3.25 times compared to the amount or level of macrophage biomarker in the reference sample, reference cells, reference tissue, control sample, control cells, or control tissue.

[0236] In some examples, a quantity or level of macrophage biomarker lower than the reference quantity or level refers to an overall decrease of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% or more compared to a pre-assigned quantity or level of macrophage biomarker detected by standard methods known in the art, such as those described herein. In specific examples, a quantity or level of macrophage biomarker lower than the reference quantity or level refers to a decrease in the quantity or level of macrophage biomarker in a sample, where the decrease is at least about 1.5 times, 1.75 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 25 times, 50 times, 75 times, or 100 times the pre-assigned quantity or level of macrophage biomarker. In some cases, a macrophage biomarker amount or level lower than the reference amount or level refers to an overall decrease in the macrophage biomarker amount or level that is greater than approximately 1.5 times, 1.75 times, 2 times, 2.25 times, 2.5 times, 2.75 times, 3.0 times, or 3.25 times compared to the macrophage biomarker pre-assigned amount or level.

[0237] (ii) Increase in macrophage biomarkers As described in International Publication No. 2022 / 031749, previous studies have determined that a patient can be identified, diagnosed, and / or predicted to benefit from treatment containing anti-CD20 antibodies if the amount or level of a macrophage biomarker in a patient's sample exceeds the amount or level of a reference macrophage biomarker. Based on the studies described herein, the amount or level of macrophage biomarkers in samples from patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) exceeding the amount or level of reference macrophage biomarkers may identify, diagnose, and / or predict that a patient may benefit from treatment comprising either an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), or an anti-CD20 antibody (e.g., obinutuzumab or rituximab). When a patient's sample has a macrophage biomarker quantity or level exceeding that of a reference macrophage biomarker, the selection of a therapy including an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), or a therapy including an anti-CD20 antibody (e.g., obinutuzumab or rituximab), may be based on additional factors in addition to the macrophage biomarker quantity or level.

[0238] (iii) Reference macrophage biomarkers The reference macrophage biomarker quantity or level may be a pre-assigned macrophage biomarker quantity or level. In some cases, the macrophage biomarker quantity or level in the reference population is the median of the macrophage biomarker quantity or level in the reference population. In some cases, the macrophage biomarker quantity or level in the reference population is the mean of the macrophage biomarker quantity or level in the reference population.

[0239] In some cases, the amount or level of a pre-assigned macrophage biomarker is the percentage of cell subtypes in the sample. In some cases, the percentage of cell subtypes in the sample is approximately 0% to 40% (e.g., 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40%). In some cases, the percentage of cell subtypes in a sample is approximately 0% to 10% (e.g., 0%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%). In some cases, the percentage of cell subtypes in a sample is less than 10% (e.g., 0%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%). In some cases, the percentage of cell subtypes in a sample is approximately 6%. In some cases, the percentage of cell subtypes in the sample is approximately 5%. In some cases, the percentage of cell subtypes in the sample is approximately 4.74%. In some cases, the percentage of cell subtypes in the sample is approximately 4%. In some cases, the percentage of cell subtypes in the sample is approximately 3.35%. In some cases, the percentage of cell subtypes in the sample is approximately 3%. In some cases, the percentage of cell subtypes in the sample is approximately 2.5%. In some cases, the percentage of cell subtypes in the sample is approximately 2%. In some cases, the percentage of cell subtypes in the sample is approximately 1.67%. In some cases, the percentage of cell subtypes in the sample is approximately 1%. In some cases, the percentage of cell subtypes in the sample is approximately 0%.

[0240] The reference doses or levels of macrophage biomarkers described herein may be based on the amount or level of macrophage biomarkers in a reference population. In some examples, the reference macrophage biomarkers described herein are the amount or level of macrophage biomarkers in a reference population that includes one or more subsets (e.g., one, two, three, four, or five) of patients.

[0241] In some examples, the reference macrophage biomarker is the quantity or level of macrophage biomarkers in a reference population, the reference population comprising at least one subset of patients having lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)).

[0242] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population comprising at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have received one or more doses (e.g., at least one, two, three, four, five, six, seven, eight, nine, or ten or more doses) of either an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0243] In some cases, the reference macrophage biomarker is the quantity or level of macrophage biomarkers in a reference population, the reference population includes at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab) as monotherapy.

[0244] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population including at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) as a combination therapy (e.g., anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) and additional therapeutic agents (e.g., anticancer therapy (e.g., cytotoxic agents, growth inhibitors, radiotherapy, anti-angiogenic agents, or combinations thereof), e.g., CHP).

[0245] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population including at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab) as part of a combination therapy (e.g., anti-CD20 antibody (e.g., obinutuzumab or rituximab) and additional therapeutic agents (e.g., anticancer therapy (e.g., cytotoxic agents, growth inhibitors, radiotherapy, anti-angiogenic agents, or combinations thereof), e.g., CHOP).

[0246] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population includes at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with anti-cancer therapies (e.g., cytotoxic agents, growth inhibitors, radiotherapy, anti-angiogenic agents, or combinations thereof), e.g., CHP, but without anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

[0247] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population includes at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with anti-cancer therapy (e.g., cytotoxic agents, growth inhibitors, radiotherapy, anti-angiogenic agents, or combinations thereof), e.g., CHOP, and who have not received anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

[0248] For example, in some cases, the reference population includes a first subset of patients who have been treated with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), and a second subset of patients who have not been treated with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0249] For example, in some cases, the reference population includes a first subset of patients who have been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab) and a second subset of patients who have not been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0250] For example, in some cases, the reference population includes a first subset of patients who have been treated with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), and a second subset of patients who have been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0251] In some cases, baseline levels or doses of macrophage biomarkers significantly separate each of the first and second subsets of patients based on the significant difference between patient responsiveness (e.g., PFS or OS) to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) and patient responsiveness to treatment with anti-CD20 antibodies (e.g., obinutuzumab or rituximab) below baseline macrophage biomarker levels, and patient responsiveness to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) is significantly improved compared to patient responsiveness to treatment with anti-CD20 antibodies (e.g., obinutuzumab or rituximab). For example, in some cases, the baseline dose or level of macrophage biomarkers optimally separates each of the first and second subsets of patients based on the maximum difference between the patient's response (e.g., PFS or OS) to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) and the patient's response below baseline macrophage biomarkers to treatment with anti-CD20 antibodies (e.g., obinutuzumab or rituximab), and the patient's response to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) is significantly improved compared to the patient's response to treatment with anti-CD20 antibodies (e.g., obinutuzumab or rituximab). In some cases, the first subset of the aforementioned patients is those treated with Pola-R-CHP (polatuzumab vedotin + rituximab, cyclophosphamide, doxorubicin, and prednisone). In some cases, the second subset of the aforementioned patients is those treated with R-CHOP (rituximab + cyclophosphamide, doxorubicin, vincristine, and prednisone).

[0252] In some cases, the baseline dose or level of macrophage biomarkers is lower than the baseline macrophage biomarkers for treatment without anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab), compared to the patient's response (e.g., PFS or OS) to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab). Based on significant differences in patient responsiveness, the first and second subsets of patients were significantly separated, and the patient responsiveness to treatment with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) was significantly improved compared to the patient responsiveness to treatment without an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab). For example, in some cases, the baseline dose or level of macrophage biomarkers is used to compare patient response (e.g., PFS or OS) to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) with baseline macrophage biomarkers for treatment without anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab). Based on the maximum difference between the patient responsiveness and the lower responsiveness, each of the first and second subsets of patients is optimally isolated, and the patient responsiveness to treatment with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) is significantly improved compared to the patient responsiveness to treatment without an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab). In some cases, the first subset of patients mentioned above consists of patients treated with Pola-R-CHP (polatuzumab vedotin + rituximab, cyclophosphamide, doxorubicin, and prednisone).

[0253] In some cases, the baseline dose or level of macrophage biomarkers significantly separates each of the first and second subsets of patients based on the significant difference between the patient's response (e.g., PFS or OS) to treatment with an anti-CD20 antibody (e.g., obinutuzumab or rituximab) and the patient's response to treatment without an anti-CD20 antibody (e.g., obinutuzumab or rituximab) with a baseline macrophage biomarker level below baseline, and the patient's response to treatment without an anti-CD20 antibody (e.g., obinutuzumab or rituximab) is significantly improved compared to the patient's response to treatment with an anti-CD20 antibody (e.g., obinutuzumab or rituximab). For example, in some cases, the baseline dose or level of macrophage biomarkers is optimally separated into first and second subsets of patients based on the maximum difference between the patient's response to treatment with an anti-CD20 antibody (e.g., obinutuzumab or rituximab) (e.g., PFS or OS) and the patient's response to treatment without an anti-CD20 antibody (e.g., obinutuzumab or rituximab) with a baseline macrophage biomarker, where the patient's response to treatment without an anti-CD20 antibody (e.g., obinutuzumab or rituximab) is significantly improved compared to the patient's response to treatment with an anti-CD20 antibody (e.g., obinutuzumab or rituximab). In some cases, the first subset of patients mentioned above is those treated with R-CHOP (rituximab + cyclophosphamide, doxorubicin, vincristine, and prednisone).

[0254] In some cases, optimal or significant separation can be based on hazard ratios (HRs) determined from an analysis of the amount or levels of macrophage biomarkers in first and second subsets of patients, where the upper limit of the 95% confidence interval for the HR is less than 1, for example, HRs of approximately 0.95, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, and 0.1 or less.

[0255] Additionally or alternatively, the reference macrophage biomarker may be the quantity or level of macrophage biomarkers in the reference population, the reference population comprising at least one subset of patients who do not have lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) or who have lymphoma but have not received treatment.

[0256] (iv) Indications The methods described herein are useful for predicting the therapeutic response of individuals with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

[0257] In some cases, the lymphoma may be a low-grade lymphoma. In some cases, the lymphoma may be a B-cell lymphoma, such as a non-Hodgkin lymphoma, such as a diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma). In some cases, the lymphoma may be a CD20-positive lymphoma.

[0258] In certain cases, cancer may be a B-cell lymphoma. For example, a B-cell lymphoma may be a non-Hodgkin lymphoma and includes, but is not limited to, diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma). For example, the methods described herein may be used to identify, diagnose, and / or predict whether a patient with B-cell lymphoma (e.g., non-Hodgkin lymphoma (e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) can benefit from treatment including an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), and the method comprises measuring macrophage biomarkers in a sample from the patient, and by a quantity or level of macrophage biomarkers in the sample being below the quantity or level of reference macrophage biomarkers, the patient is identified, diagnosed, and / or predicted to benefit from treatment including an anti-CD79b immunoconjugate and an anti-CD20 antibody.

[0259] In some cases, individuals with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) have never been treated for lymphoma before (treatment-inexperienced). For example, in some cases, individuals with lymphoma have never been previously administered either an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) or an anti-CD20 antibody (e.g., obinutuzumab or rituximab) or an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0260] In some cases, individuals with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) have previously received treatment for lymphoma. In some cases, individuals with lymphoma have previously received treatment including either an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) or an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0261] (v) Therapeutic benefits Patients who benefit from treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) may experience, for example, delay or prevention of the onset or recurrence of lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)), relief of symptoms, reduction of the direct or indirect pathological outcomes of cancer, prevention of metastasis, slower rate of disease progression, improvement or relief of symptoms, or remission or improved prognosis. In some cases, the treatments described herein are used to delay the onset of cancer or to delay the progression of lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)). In some cases, the benefit may be an increase in overall survival (OS), progression-free survival (PFS), complete remission (CR), partial remission (PR), or a combination thereof.

[0262] In some cases, individuals are identified as those who would benefit from treatment including anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) if the amount or level of macrophage biomarkers is below the amount or level of reference macrophage biomarkers (e.g., the amount or level of macrophage biomarkers in the reference population), where the benefit is defined as anti-CD79b immunoconjugates. The increase in overall survival (OS) compared to treatment without the combination (e.g., polatuzumab vedotin) and anti-CD20 antibody (e.g., obinutuzumab or rituximab) is (e.g., ≥20%, ≥25%, ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%).

[0263] In some cases, a macrophage biomarker amount or level that is below the reference amount or level of macrophage biomarker (e.g., the amount or level of macrophage biomarker in the reference population) identifies an individual as someone who would benefit from treatment including an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), where the benefit is the anti-CD79b immunoconjugate ( For example, an increase in PFS (e.g., ≥20%, ≥25%, ≥30%, ≥35%, ≥40%, ≥45%, ≥50%, ≥55%, ≥60%, ≥65%, ≥70%, ≥75%, ≥80%, ≥85%, ≥90%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) compared to treatment without polatuzumab vedotin and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0264] B. Determination of macrophage biomarkers (i) Detection method The macrophage biomarkers described herein may be based on the quantity or expression level of nucleic acids (e.g., mRNA), proteins, or cells (e.g., M1 macrophages or tumor-associated macrophages). The presence and / or expression level / quantity of genes described herein (see, for example, Table 2 or Table 3) can be determined qualitatively and / or quantitatively based on any appropriate criteria known in the art, including but not limited to DNA, mRNA, cDNA, proteins, protein fragments, and / or gene copy number. The presence and / or level / quantity of cells described herein can be determined qualitatively and / or quantitatively based on any appropriate criteria known in the art, including but not limited to microscopy, cytometry, DNA, mRNA, cDNA, proteins, protein fragments, and / or gene copy number. [Table 2] [Table 3]

[0265] In some cases, the nucleic acid expression levels of the genes described herein can be measured by polymerase chain reaction (PCR)-based assays, such as quantitative PCR, real-time PCR, quantitative real-time PCR (qRT-PCR), reverse transcriptase PCR (RT-PCR), and reverse transcriptase quantitative PCR (RT-qPCR). A platform for performing quantitative PCR assays is Fluidigm (e.g., BIOMARK® HD System). Other amplification-based methods include, for example, transcription-mediated amplification (TMA), strand-displacement amplification (SDA), nucleic acid-based amplification (NASBA), and signal amplification methods, such as bDNA.

[0266] In some cases, the nucleic acid expression levels of the genes described herein may also be measured by sequencing-based techniques such as RNA-seq, sequential gene expression analysis (SAGE), high-throughput sequencing techniques (e.g., large-scale parallel sequencing), and Sequenom MassARRAY®. Nucleic acid expression levels may also be measured by, for example, NanoString nCounter and high-coverage expression profiling (HiCEP). Additional protocols for evaluating the state of genes and gene products can be found, for example, in Part 2 (Northern blotting), Part 4 (Southern blotting), Part 15 (Immunoblotting), and Part 18 (PCR analysis) of Ausubel et al., eds., 1995, Current Protocols In Molecular Biology.

[0267] Other methods for detecting the nucleic acid level of genes described herein include protocols for examining or detecting mRNA, such as target mRNA, in tissue or cell samples using microarray technology. Using nucleic acid microarrays, test and control mRNA samples from test and control tissue samples are reverse transcribed and labeled to generate cDNA probes. The probes are then hybridized to a nucleic acid array immobilized on a solid support. The array is configured so that the sequence and position of each member of the array are known. Hybridization of a labeled probe with a specific array member indicates that the sample from which the probe originates expresses that gene.

[0268] Primers and probes may be labeled with detectable markers such as radioisotopes, fluorescent compounds, bioluminescent compounds, chemiluminescent compounds, metal chelators, or enzymes. Such probes and primers can be used to detect the presence of expressed genes (e.g., genes described herein). As those skilled in the art will understand, many different primers and probes can be prepared based on the sequences provided herein (or, in the case of genomic DNA, their adjacent sequences) and used to effectively amplify, clone, and / or determine the presence and / or expression levels of the genes described herein.

[0269] Other methods for detecting the nucleic acid expression levels of the genes described herein include electrophoresis, Northern blotting and Southern blotting, in situ hybridization (e.g., single or multiple nucleic acid in situ hybridization), RNAse protection assays, and microarrays (e.g., Illumina BEADARRAY® technology, bead arrays for detecting gene expression (BADGE)).

[0270] In some cases, macrophage biomarkers can be analyzed by a wide range of methodologies, including but not limited to RNA-seq, PCR, RT-qPCR, qPCR, multiplex qPCR, multiplex RT-qPCR, NANOSTRING® nCOUNTER® gene expression assays, microarray analysis, sequential gene expression analysis (SAGE), Northern blotting, MassARRAY, ISH, whole-genome sequencing, FACS, spatial transcriptomics, spatial proteomics, Western blotting, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blotting, immunodetection, surface plasmon resonance, optical spectroscopy, mass spectrometry, and HPLC, or combinations thereof.

[0271] (ii) RT-qPCR In some cases, the nucleic acid expression levels of genes described herein (e.g., genes in the M1 macrophage gene signature set, genes in the tumor-associated macrophage gene signature set, or genes in the gene signature matrix) can be detected using reverse transcription quantitative polymerase chain reaction (RT-qPCR). The RT-qPCR technique is a form of PCR in which the nucleic acid to be amplified is first RNA reverse-transcribed into cDNA, and the amount of PCR product is measured at each step of the PCR reaction. Since RNA cannot be used as a PCR template, the first step in PCR gene expression profiling is the reverse transcription of the RNA template into cDNA, followed by its amplification in the PCR reaction. For example, reverse transcriptases may include avilo myeloblastosis virus reverse transcriptase (AMY-RT) or Moloney mouse leukemia virus reverse transcriptase (MMLV-RT). The reverse transcription step is typically primed using specific primers, random hexamers, or oligo-dT primers, depending on the context and the goal of expression profiling. For example, the extracted RNA can be reverse transcribed using the GENEAMP® RNA PCR kit (Perkin Elmer, California, USA) according to the manufacturer's instructions. The resulting cDNA can then be used as a template in subsequent PCR reactions.

[0272] One variation of the PCR technique is quantitative real-time PCR (qRT-PCR), which measures the accumulation of PCR products through a double-labeled fluorescent probe (i.e., a TAQMAN® probe). Quantitative real-time polymerase chain reaction (QRT-PCR) techniques refer to a form of PCR in which the amount of PCR product is measured at each step of the PCR reaction. This technique has been described in various publications, including Cronin et al., Am.J. Pathol. 164(l):35-42 (2004) and Ma et al., Cancer Cell 5:607-616 (2004). Real-time PCR is compatible with quantitative competitive PCR, which uses competitors within each target sequence for normalization, and / or quantitative comparative PCR, which uses normalization genes contained within the sample or housekeeping genes for PCR. For further details, see, for example, Held et al., Genome Research 6:986-994 (1996).

[0273] The steps of a typical protocol for profiling gene expression using fixed paraffin-embedded tissue as an RNA source, such as mRNA isolation, purification, primer extension, and amplification, are provided in various published journal articles (e.g., Godfrey et al., Malec. Diagnostics 2:84-91 (2000), Specht et al., Am. J. Pathol. 158:419-29 (2001)). Briefly, a typical process begins with cutting sections of paraffin-embedded tumor tissue samples (e.g., 10-microgram sections). RNA is then extracted, and proteins and DNA are removed. After analyzing the RNA concentration, RNA repair and / or amplification steps may be included as needed, where the RNA is reverse transcribed using a gene-specific promoter, and then PCR is performed.

[0274] Nucleic acid expression levels determined by amplification-based methods (e.g., RT-qPCR) can be expressed as a cycle threshold (Ct). From this value, the normalized expression level of each gene can be determined using a delta-Ct (dCt) method, for example, as follows: Ct(control / reference gene) - Ct(desired gene / target gene) = dCt(desired gene / target gene). Those skilled in the art will understand that the obtained dCt value can be negative or positive. As defined herein, a higher dCt value indicates a higher expression level of the desired gene compared to the control gene. Conversely, a lower dCt value indicates a lower expression level of the desired gene compared to the control gene. When the expression levels of multiple genes have been determined, the expression levels of each gene, expressed as dCt values, can be used to determine a single value representing the aggregated or composite expression level of multiple genes (e.g., genes in the M1 macrophage gene signature set or genes in the TAM gene signature set). The aggregated or combined expression level may be the mean or median dCt values ​​determined for each target gene / target gene. As defined herein, a higher mean or median dCt indicates a higher aggregate expression level for multiple target genes compared to a control gene (or multiple control genes). A lower mean or median dCt indicates a lower aggregate expression level for multiple target genes compared to a control gene (or multiple control genes). Expression levels can be compared to baseline levels.

[0275] In a specific example, the nucleic acid expression levels described herein may be determined using a method including the following: (a) obtaining or providing a sample from an individual (the sample includes a tumor tissue sample (e.g., a paraffin-embedded formalin-fixed tumor tissue sample)), (b) isolating mRNA from the sample, (c) reverse transcribing the mRNA into cDNA (for example, for at least one of the genes described herein (e.g., a gene in the M1 macrophage gene signature set, a gene in the TAM gene signature set, or a gene in the gene signature matrix)), (d) amplifying the cDNA (for example, for at least one of the genes described herein (e.g., a gene in the M1 macrophage gene signature set, a gene in the TAM gene signature set, or a gene in the gene signature matrix)) using PCR, and (e) quantifying the nucleic acid expression level (for example, for at least one of the genes described herein (e.g., a gene in the M1 macrophage gene signature set, a gene in the tumor-associated macrophage (TAM) gene signature set, or a gene in the gene signature matrix)).

[0276] Depending on the primer or probe used, one or more genes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more genes (e.g., 55, 82, 89, 106, 153, or 170 genes)) can be detected in a single assay. Furthermore, the assay can be performed across one or more tubes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tubes (e.g., 55, 82, 89, 106, 153, or 170 tubes)).

[0277] In some examples, the method further includes (f) normalizing the nucleic acid expression level of a gene in the sample (e.g., at least one of the genes described herein (e.g., a gene in the M1 macrophage gene signature set, a gene in the tumor-associated macrophage (TAM) gene signature set, or a gene in the gene signature matrix)) to the expression level of one or more reference genes (e.g., one, two, three, four, five, six, seven, eight, nine or more reference genes, e.g., housekeeping genes (e.g., β-actin)). For example, RT-qPCR can be used to analyze the expression levels of the genes described herein (e.g., at least one of the genes described herein (e.g., a gene in the M1 macrophage gene signature set, a gene in the tumor-associated macrophage gene signature set, or a gene in the gene signature matrix)) to generate an expression level that reflects the normalized mean dCT value of the analyzed gene.

[0278] (iii) RNA-seq and microarrays In some cases, the nucleic acid expression levels of genes described herein (e.g., genes in the M1 macrophage gene signature set, genes in the TAM gene signature set, or genes in the gene signature matrix) can be detected using RNA-seq. RNA-seq, also known as Whole Transcriptome Shotgun Sequencing (WTSS), refers to the use of high-throughput sequencing techniques to sequence and / or quantify cDNA to obtain information about the RNA content of a sample. Publications describing RNA-Seq include Wang et al., “RNA-Seq: a revolutionary tool for transcriptomics,” Nature Reviews Genetics 10(1):57-63 (January 2009), Ryan et al., BioTechniques 45(1):81-94 (2008), and Maher et al., “Transcriptome sequencing to detect gene fusions in cancer.” Nature 458(7234):97-101 (January 2009). In some cases, quality control is performed on the sequencing. In some cases, the count is normalized to transcript permillions (TPM).

[0279] (a) Marker gene approach The marker gene approach uses the expression of one or more genes within a set of gene signatures (see, for example, Tables 2 and 3) to determine macrophage biomarkers (e.g., the number of M1 macrophages or tumor-associated macrophages in a sample). In some examples, the marker gene approach uses xCell (see, for example, Aran et al. Genome Biol. 18(1):220 (2017)).

[0280] The gene signature sets illustrated in Tables 2 and 3 can be modified to remove, replace, or add genes. In some examples, the number of genes in any of the illustrated gene signature sets can be increased or decreased by one or more genes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more genes). In some examples, the number of genes in any of the illustrated gene signature sets can be increased or decreased by approximately 5% to approximately 20% (e.g., 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%). In some examples, a gene from any of the illustrated gene signature sets can be replaced by a gene in the same signaling pathway. In some examples, a gene signature set can be generated by adding genes from one of the exemplary gene signature sets to a different exemplary gene signature set (for example, a gene from gene signature set 1 (e.g., C1QA) can be added to gene signature set 2). In some examples, a gene signature set may contain genes present in all exemplary gene signature sets (i.e., ACP2 and ADAMDEC1) or most exemplary gene signature sets (e.g., FDX1, CD163, HAMP, ABCD1, C1QA, CCL22, and TREM2). In some examples, a gene signature set contains ACP2 and ADAMDEC1. In some examples, a gene signature set contains ACP2, ADAMDEC1, and FDX1. In some examples, a gene signature set contains ACP2, ADAMDEC1, and CD163. In some examples, the gene signature set includes ACP2, ADAMDEC1, and HAMP. In some examples, the gene signature set includes ACP2, ADAMDEC1, FDX1, and CD163. In some examples, the gene signature set includes ACP2, ADAMDEC1, FDX1, and HAMP. In some examples, the gene signature set includes ACP2, ADAMDEC1, CD163, and HAMP.In some examples, the gene signature set includes ACP2, ADAMDEC1, FDX1, CD163, and HAMP. In some examples, the gene signature set includes ACP2, ADAMDEC1, FDX1, CD163, HAMP, and ABCD1. In some examples, the gene signature set includes ACP2, ADAMDEC1, FDX1, CD163, HAMP, and CCL22. In some examples, the gene signature set includes ACP2, ADAMDEC1, FDX1, CD163, HAMP, and C1QA. In some examples, the gene signature set includes ACP2, ADAMDEC1, FDX1, CD163, HAMP, and TREM2. In some examples, the gene signature set includes ACP2, ADAMDEC1, FDX1, CD163, HAMP, ABCD1, and CCL22.

[0281] A representative protocol for determining the number of macrophages (e.g., M1 macrophages or tumor-associated macrophages) using an RNA-seq marker gene approach can be found in Aran et al. Genome Biol. 18(1):220 (2017). Briefly, sequence scores obtained from a sample (e.g., pre-treated raw sequence reads) are transformed based on a function derived from a synthetic mixture of cell types (e.g., M1 macrophages or tumor-associated macrophages) and control cell types (e.g., pluripotent progenitor cells or endothelial cells) within a range (e.g., 0.8% to 25.6%) based on the expected abundance of each cell type present in the sample. The transformed scores are further refined using a spillover compensation matrix (restricted to 0.5 from the diagonal) derived from a synthetic mixture of 25% cell types (e.g., M1 macrophages or tumor-associated macrophages) and 75% control cell types (e.g., pluripotent progenitor cells or endothelial cells). The final refined scores represent the proportion of each cell type present in the sample.

[0282] (b) Deconvolution approach The deconvolution approach uses the expression of one or more genes in a gene signature matrix to determine macrophage biomarkers (e.g., the number of M1 macrophages or tumor-associated macrophages in a sample). In some cases, the deconvolution approach uses quanTIseq (see, e.g., Finotello et al. Genome Med. 11(1):34(2019)).

[0283] A representative protocol for determining the number of M1 macrophages or tumor-associated macrophages using an RNA-seq deconvolution approach can be found in Finotello et al. Genome Med. 11(1):34 (2019). Briefly, the sequence scores obtained from the sample (e.g., pre-treated raw sequence reads) are normalized, deconvolved using a gene signature matrix, and then constrained least-squares regression is used to determine the cell types present in the sample (e.g., M1 macrophages, M2 macrophages, tumor-associated macrophages, B cells, monocytes, neutrophils, NK cells, unregulated CD4 cells). + T cells, CD8 + Calculate the proportion of T cells, regulatory T cells, dendritic cells, or other cell types.

[0284] (iv) Immunohistochemistry In some cases, macrophages (e.g., M1 macrophages or tumor-associated macrophages) can be detected using immunohistochemistry (IHC). In some cases, macrophages (e.g., M1 macrophages or tumor-associated macrophages) can be identified or distinguished from other cell types using any of the genes described herein (e.g., CD68, genes in the M1 macrophage gene signature set, genes in the tumor-associated macrophage gene set, or genes in the gene signature matrix). In some cases, an antibody specific to any of the genes described herein (e.g., CD68, genes in the M1 macrophage gene signature set, genes in the tumor-associated macrophage gene signature set, or genes in the gene signature matrix) is used as the primary antibody in the IHC assay. In some cases, a horseradish peroxidase (HRP) conjugate secondary antibody is used in the IHC assay. In some cases, the signal from the IHC assay is compared to an IHC assay performed using a negative control antibody. In some cases, macrophages (e.g., M1 macrophages or tumor-associated macrophages) can be detected using antibodies that bind to CD68.

[0285] (v) Flow cytometry In some cases, macrophages (e.g., M1 macrophages or tumor-associated macrophages) can be detected using flow cytometry. In some cases, macrophages (e.g., M1 macrophages or tumor-associated macrophages) can be identified or distinguished from other cell types using any of the genes described herein (e.g., genes in the M1 macrophage gene signature set, genes in the tumor-associated macrophage gene set, or genes in the gene signature matrix). In some cases, macrophages (e.g., M1 macrophages or tumor-associated macrophages) can be labeled using antibodies specific to any of the genes described herein (e.g., genes in the M1 macrophage gene signature set, genes in the tumor-associated macrophage gene signature set, or genes in the gene signature matrix).

[0286] (vi) Sample Samples may be collected from individuals suspected of having lymphoma, or diagnosed with lymphoma, and therefore likely to require treatment, or from healthy individuals not suspected of having lymphoma, or who do not have lymphoma but have a family history of lymphoma. In the case of gene expression assessment, samples such as cells, or those containing proteins or nucleic acids produced by these cells, may be used in the method of the present invention. Gene expression levels can be determined by evaluating the amount (e.g., absolute amount or concentration) of markers in the sample (e.g., tissue samples, tumor tissue samples such as biopsy samples). In addition, gene levels can be evaluated in bodily fluids or secretions containing detectable levels of genes. Examples of bodily fluids or secretions useful as samples in the present invention include, for example, blood, urine, saliva, feces, pleural fluid, lymph, sputum, ascites, prostatic fluid, cerebrospinal fluid (CSF), or any other bodily secretions or their derivatives. The term blood is intended to include whole blood, plasma, serum, or any blood derivatives. Such assessment of genes in bodily fluids or secretions may be preferable in situations where invasive sampling methods are inappropriate or inconvenient. In other embodiments, tumor tissue samples are preferred.

[0287] Samples may be frozen, kept fresh, fixed (e.g., formalin fixation), centrifuged, and / or embedded (e.g., paraffin embedding). Cell samples may be subjected to various well-known post-collection preparation and storage techniques (e.g., nucleic acid and / or protein extraction, fixation, storage, freezing, ultrafiltration, concentration, evaporation, centrifugation, etc.) prior to evaluating the amount of markers in the sample. Similarly, biopsies may be subjected to post-collection preparation and storage techniques, such as fixation by formalin fixation.

[0288] In one specific example, the sample is a clinical sample. In another example, the sample is used in a diagnostic assay, such as the diagnostic assay or diagnostic method of the present invention. In some examples, the sample is obtained from a primary or metastatic tumor. Tissue biopsy is often used to obtain a representative small piece of tumor tissue. Alternatively, tumor cells may be obtained indirectly in the form of tissue or fluid known or thought to contain the tumor cells of interest. For example, a sample of a lymphoma lesion may be obtained by excision, fine-needle aspiration, pleural fluid, or blood. Genes or gene products can be detected from cancer or tumor tissue, or from other body samples, such as urine, sputum, serum, or plasma. The same techniques discussed above for the detection of target genes or gene products in cancerous samples may be applied to other body samples. Cancer cells may detach from cancerous lesions and appear in such body samples. By screening such body samples, a simple early diagnosis of these cancers can be achieved. In addition, the progress of therapy can be more easily monitored by testing such body samples for target genes or gene products.

[0289] In some cases, individual-derived samples are tissue samples, whole blood samples, plasma samples, serum samples, or combinations thereof. In some cases, the sample is a tissue sample. In some cases, the sample is a tumor tissue sample. In some cases, the sample is obtained before treatment. In some cases, the tissue sample is a formalin-fixed and paraffin-embedded (FFPE) sample, an archived sample, a fresh sample, or a frozen sample. In some cases, individual-derived samples are tissue samples. In some cases, the tissue sample is a tumor tissue sample (e.g., a biopsy sample). In some cases, the tumor tissue sample contains tumor cells, tumor-infiltrating immune cells, stromal cells, normal adjacent tissue (NAT) cells, or combinations thereof. In some cases, the tissue sample is a biopsy sample. In some cases, the tissue sample is a blood cell, lymph node, or bone / bone marrow sample.

[0290] In some cases, tumor tissue samples are extracted from malignant cancerous tumors (i.e., cancer). In some cases, the cancer is a solid tumor, or a non-solid or soft tissue tumor. In some cases, the tumor tissue sample is from a lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)).

[0291] (vii) RNA extraction Prior to detecting nucleic acid levels, mRNA can be isolated from the target sample. In some cases, mRNA is total RNA isolated from a tumor or tumor cell line, or alternatively, from normal tissue or cell line. When the mRNA source is a primary tumor, mRNA can be extracted, for example, from frozen or stored, paraffin-embedded and fixed (e.g., formalin-fixed) tissue samples. General methods for mRNA extraction are well known in the art and are disclosed in standard molecular biology textbooks, including Ausubel et al., *Current Protocols of Molecular Biology*, John Wiley and Sons (1997). Methods for RNA extraction from paraffin-embedded tissue are disclosed, for example, in Rupp and Locker, *Lab Invest. 56:A67* (1987) and De Andres et al., *Bio Techniques* 18:42044 (1995). In particular, RNA isolation can be carried out using purification kits, buffer sets, and proteases from commercial manufacturers such as Qiagen, according to the manufacturer's instructions. For example, total RNA derived from cells in a culture can be isolated using a Qiagen RNeasy mini-column. Other commercially available RNA isolation kits include the MASTERPURE® Complete DNA and RNA Purification Kit (EPICENTRE®, Madison, Wisconsin) and the Paraffin Block RNA Isolation Kit (Ambion, Inc.). Total RNA derived from tissue samples can be isolated, for example, using RNA Stat-60 (TelTest). RNA prepared from tumor tissue samples can also be isolated, for example, by cesium chloride density gradient centrifugation.

[0292] (viii) Expression level The expression levels may reflect the expression levels of one or more genes described herein (e.g., one or more genes in the M1 macrophage gene signature set, one or more genes in the tumor-associated macrophage gene signature set, or one or more genes in the gene signature matrix). In certain examples, the detected expression level of each gene is normalized using one of the standard normalization methods known in the art. Those skilled in the art will understand that the normalization method used may depend on the gene expression methodology used (e.g., one or more housekeeping genes may be used for normalization from the perspective of the RT-qPCR methodology, while the whole genome or substantially the whole genome may be used as the normalization baseline from the perspective of the RNA-seq methodology). For example, the detected expression level of each assayed gene may be normalized for differences in the quantity of assayed genes, variability in the quality of the samples used, and / or variability between assay runs.

[0293] In some cases, normalization can be achieved by detecting the expression of one or more specific normalization genes, including reference genes (e.g., housekeeping genes (e.g., β-actin)). For example, in some cases, nucleic acid expression levels detected using the methods described herein (e.g., for at least one of the genes described herein (e.g., genes in the M1 macrophage gene signature set, genes in the tumor-associated macrophage gene signature set, or genes in the gene signature matrix)) can be normalized to the expression levels of one or more reference genes (e.g., one, two, three, four, five, six, seven, eight, nine or more reference genes, e.g., housekeeping genes (e.g., β-actin)). Alternatively, normalization can be performed based on the mean or median of signals from all assayed genes. For each gene, the measured amount of normalized mRNA can be compared to the amount found at the reference expression level. The measured presence and / or expression level / amount in the specific target sample being analyzed is expressed at a percentile within this range, which can be determined by methods well known in the art.

[0294] In other cases, the detected expression levels of each assayed gene are not normalized in order to determine the expression level.

[0295] The expression levels may reflect the aggregated or combined expression levels of a single gene or multiple genes described herein (for example, for at least one of the genes described herein (e.g., genes in the M1 macrophage gene signature set, genes in the tumor-associated macrophage gene signature set, or genes in the gene signature matrix)). The expression levels can be determined using any statistical method known in the art.

[0296] For example, the expression level may reflect a numerical value that reflects the median, mean, or aggregated Z-score expression level for a combination of assayed genes (for example, for at least one of the genes described herein (e.g., genes in the M1 macrophage gene signature set, genes in the tumor-associated macrophage gene signature set, or genes in the gene signature matrix)).

[0297] In some examples, the expression level reflects the median normalized expression level, the mean normalized expression level, or a numerical value that reflects the aggregated Z-score normalized expression level for the combination of genes being assayed (for example, for at least one of the genes described herein (e.g., a gene in the M1 macrophage gene signature set, a gene in the tumor-associated macrophage gene signature set, or a gene in the gene signature matrix)).

[0298] IV. Treatment methods, compositions and uses Methods, compositions, and uses thereof for treating patients having lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) are provided herein, the method comprising administering an effective amount of anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and anti-CD20 antibody (e.g., obinutuzumab or rituximab) to a patient based on a sample from the patient (e.g., tissue sample, e.g., tumor tissue sample, e.g., biopsy sample) determined by a macrophage biomarker (e.g., gene expression value (e.g., gene expression value derived from any of the gene signature sets described herein (e.g., any of the exemplary gene signature sets in Table 2 or Table 3)) or macrophages (e.g., M1 macrophages or tumor-associated macrophages)).

[0299] In some cases, anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) may be administered as first-line treatment. Alternatively, anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) may be administered as second-line treatment.

[0300] In any of the following sections, the lymphoma may be a B-cell lymphoma. In some cases, the B-cell lymphoma is a non-Hodgkin lymphoma. In preferred embodiments, the non-Hodgkin lymphoma is DLBCL. In some cases, DLBCL is germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma. In some cases, the lymphoma is a low-grade lymphoma. In some cases, the lymphoma is a CD20-positive lymphoma.

[0301] In all of the following sections, the patient is human. In some cases, the patient has never been treated before. In some cases, the patient has been treated before. In some cases, the patient has been previously treated with either an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) or an anti-CD20 antibody. In some cases, the patient has never been previously treated with either an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) or an anti-CD20 antibody.

[0302] In any of the following sections, the sample may be a tissue sample, a tumor sample, a whole blood sample, a plasma sample, or a serum sample. In some examples, the tissue sample is a tumor tissue sample. In some examples, the tumor tissue sample includes tumor cells, tumor-infiltrating immune cells, stromal cells, normal adjacent tissue (NAT) cells, or a combination thereof. In some examples, the tumor tissue sample is a biopsy sample. In some examples, the sample is an archived sample, a fresh sample, or a frozen sample.

[0303] In any of the following sections, macrophage biomarkers can be measured directly or indirectly. In some cases, macrophage biomarkers are cells, nucleic acids, proteins, lipids, or carbohydrates. In some cases, macrophage biomarkers are gene expression levels. In some cases, macrophage biomarkers are the quantity of macrophages (e.g., M1 macrophages or tumor-associated macrophages).

[0304] A. Macrophage biomarkers for use in therapeutic methods In certain cases, a method of treating patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) with an effective dose of anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and anti-CD20 antibody (e.g., obinutuzumab or rituximab) is based on macrophage biomarkers (e.g., gene expression levels (e.g., gene expression levels derived from any of the gene signature sets described herein (e.g., any of the exemplary gene signature sets in Table 2 or Table 3)) or macrophages (e.g., M1 macrophages or tumor-associated macrophages)) determined in a sample from the patient (e.g., tissue sample, e.g., tumor tissue sample, e.g., biopsy sample).

[0305] In one embodiment, a method for treating a patient having lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) is provided herein, comprising: (a) measuring the amount of macrophage biomarkers (e.g., gene expression values ​​(e.g., gene expression values ​​derived from any of the gene signature sets described herein (e.g., any of the exemplary gene signature sets in Table 2 or Table 3)) or macrophages (e.g., M1 macrophages or tumor-associated macrophages)) in a sample from the patient (e.g., tissue sample, e.g., tumor tissue sample, e.g., biopsy sample, etc.), wherein the amount or level of macrophage biomarkers in the sample is below the amount or level of reference macrophage biomarkers; and (b) administering to the patient an effective amount of anti-CD79b immunoconjugate and anti-CD20 antibody based on the macrophage biomarkers measured in step (a).

[0306] In another embodiment, a method is provided herein for treating a patient having lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) comprising administering an effective amount of anti-CD79b immunoconjugate and anti-CD20 antibody to the patient, wherein prior to treatment, it is determined that the amount or level of macrophage biomarkers (e.g., gene expression values ​​(e.g., gene expression values ​​derived from any of the gene signature sets described herein (e.g., any of the exemplary gene signature sets in Table 2 or Table 3)) or macrophages (e.g., M1 macrophages or tumor-associated macrophages) in a sample from the patient (e.g., tissue sample, e.g., tumor tissue sample, e.g., biopsy sample) is below the amount or level of a reference macrophage biomarker.

[0307] In another embodiment, the present invention provides a method for treating a patient having lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) in which the amount or level of macrophage biomarkers (e.g., gene expression values ​​(e.g., gene expression values ​​derived from any of the gene signature sets described herein (e.g., any of the exemplary gene signature sets in Table 2 or Table 3)) or macrophages (e.g., M1 macrophages or tumor-associated macrophages) in a sample from the patient (e.g., tissue sample, e.g., tumor tissue sample, e.g., biopsy sample, etc.) is below the amount or level of a reference macrophage biomarker, comprising administering an effective amount of anti-CD79b immunoconjugate and anti-CD20 antibody to the patient.

[0308] The amounts or levels of macrophage biomarkers that determine the various methods described herein are further described below.

[0309] (i) Decrease in macrophage biomarkers If the amount or level of macrophage biomarkers in a sample from a patient with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) is below the amount or level of reference macrophage biomarkers, it may be determined that the patient should be administered an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0310] In some cases, the amount or level of macrophage biomarkers in a sample is approximately at the lower 99th percentile (prevalence level of approximately 99% or less), approximately at the lower 95th percentile (prevalence level of approximately 95% or less), approximately at the lower 90th percentile (prevalence level of approximately 90% or less), approximately at the lower 85th percentile (prevalence level of approximately 85% or less), approximately at the lower 80th percentile (prevalence level of approximately 80% or less), approximately at the lower 75th percentile (prevalence level of approximately 75% or less), approximately at the lower 70th percentile (prevalence level of approximately 70% or less), approximately at the lower 65th percentile (prevalence level of approximately 65% ​​or less), approximately at the lower 60th percentile (prevalence level of approximately 60% or less), approximately at the lower 55th percentile (prevalence level of approximately 55% or less), and approximately at the lower 50th percentile (prevalence level of approximately 50% or less). (Level), approximately the lower 45th percentile (prevalence level of approximately 45% or less), approximately the lower 40th percentile (prevalence level of approximately 40% or less), approximately the lower 35th percentile (prevalence level of approximately 35% or less), approximately the lower 30th percentile (prevalence level of approximately 30% or less), approximately the lower 25th percentile (prevalence level of approximately 25% or less), approximately the lower 20th percentile (prevalence level of approximately 20% or less), approximately the lower 15th percentile A method of treating patients who are within the centile (prevalence level of approximately 15% or less), the lower 10th percentile (prevalence level of approximately 10% or less), the lower 5th percentile (prevalence level of approximately 5% or less), or the lower 1st percentile (prevalence level of approximately 1% or less) includes administering a therapy comprising an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0311] In some cases, a method of treating a patient in whom the amount or level of a macrophage biomarker in a sample falls within the approximately lower 10th to approximately lower 90th percentile, approximately lower 20th to approximately lower 80th percentile, approximately lower 30th to approximately lower 70th percentile, approximately lower 40th to approximately lower 60th percentile, approximately lower 45th to approximately lower 55th percentile, approximately lower 48th to approximately lower 52nd percentile, approximately lower 49.5th to approximately lower 50.5th percentile, approximately lower 49.9th to approximately lower 50.1th percentile, or approximately lower 50th percentile of the amount or level of a macrophage biomarker in a reference population includes administering a therapy comprising an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab). For example, in some cases, the amount or level of macrophage biomarkers in a sample corresponds to a prevalence of approximately 10% to 90% in the reference population, approximately 15% to 85%, approximately 20% to 80%, approximately 25% to 75%, approximately 30% to 70%, approximately 35% to 65%, approximately 40% to 60%, and approximately 45% to 55%. Methods for treating patients with a prevalence of approximately 48% to 52%, approximately 49.5% to 50.5%, approximately 49.9% to 50.1%, or approximately 50% include administering a therapy comprising an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0312] In some cases, a macrophage biomarker amount or level lower than the reference amount or level refers to a decrease of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% or more compared to the amount or level of macrophage biomarker detected by standard methods known in the art, such as those described herein, compared to the amount or level of macrophage biomarker in a reference sample, reference cells, reference tissue, control sample, control cells, or control tissue. In certain cases, a macrophage biomarker amount or level lower than the reference amount or level refers to a decrease in the amount or level of macrophage biomarker in a sample, where this decrease is at least about 1.5 times, 1.75 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 25 times, 50 times, 75 times, or 100 times the amount or level of macrophage biomarker in the reference sample, reference cells, reference tissue, control sample, control cells, or control tissue. In some cases, a macrophage biomarker amount or level lower than the reference amount or level refers to a decrease in the amount or level of macrophage biomarker that is more than about 1.5 times, about 1.75 times, about 2 times, about 2.25 times, about 2.5 times, about 2.75 times, about 3.0 times, or about 3.25 times compared to the amount or level of macrophage biomarker in the reference sample, reference cells, reference tissue, control sample, control cells, or control tissue.

[0313] In some examples, a quantity or level of macrophage biomarker lower than the reference quantity or level refers to an overall decrease of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% or more compared to a pre-assigned quantity or level of macrophage biomarker detected by standard methods known in the art, such as those described herein. In specific examples, a quantity or level of macrophage biomarker lower than the reference quantity or level refers to a decrease in the quantity or level of macrophage biomarker in a sample, where this decrease is at least about 1.5 times, 1.75 times, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 25 times, 50 times, 75 times, or 100 times the pre-assigned quantity or level of macrophage biomarker. In some cases, a macrophage biomarker amount or level lower than the reference amount or level refers to an overall decrease in the macrophage biomarker amount or level that is greater than approximately 1.5 times, 1.75 times, 2 times, 2.25 times, 2.5 times, 2.75 times, 3.0 times, or 3.25 times compared to the macrophage biomarker pre-assigned amount or level.

[0314] (ii) Increase in macrophage biomarkers As described in International Publication No. 2022 / 031749, previous studies have determined that a patient can be identified, diagnosed, and / or predicted to benefit from treatment containing anti-CD20 antibodies if the amount or level of a macrophage biomarker in a patient's sample exceeds the amount or level of a reference macrophage biomarker. Based on the studies described herein, the amount or level of macrophage biomarkers in samples from patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) exceeding the amount or level of reference macrophage biomarkers may identify, diagnose, and / or predict that a patient may benefit from treatment comprising either an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), or an anti-CD20 antibody (e.g., obinutuzumab or rituximab). When a patient's sample has a macrophage biomarker quantity or level exceeding that of a reference macrophage biomarker, the selection of a therapy including an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), or a therapy including an anti-CD20 antibody (e.g., obinutuzumab or rituximab), may be based on additional factors in addition to the macrophage biomarker quantity or level.

[0315] (iii) Reference macrophage biomarkers The reference macrophage biomarker quantity or level may be a pre-assigned macrophage biomarker quantity or level. In some cases, the macrophage biomarker quantity or level in the reference population is the median of the macrophage biomarker quantity or level in the reference population. In some cases, the macrophage biomarker quantity or level in the reference population is the mean of the macrophage biomarker quantity or level in the reference population.

[0316] In some cases, the amount or level of a pre-assigned macrophage biomarker is the percentage of cell subtypes in the sample. In some cases, the percentage of cell subtypes in the sample is approximately 0% to 40% (e.g., 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40%). In some cases, the percentage of cell subtypes in a sample is approximately 0% to 10% (e.g., 0%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%). In some cases, the percentage of cell subtypes in a sample is less than 10% (e.g., 0%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%). In some cases, the percentage of cell subtypes in a sample is approximately 6%. In some cases, the percentage of cell subtypes in the sample is approximately 5%. In some cases, the percentage of cell subtypes in the sample is approximately 4.74%. In some cases, the percentage of cell subtypes in the sample is approximately 4%. In some cases, the percentage of cell subtypes in the sample is approximately 3.35%. In some cases, the percentage of cell subtypes in the sample is approximately 3%. In some cases, the percentage of cell subtypes in the sample is approximately 2.5%. In some cases, the percentage of cell subtypes in the sample is approximately 2%. In some cases, the percentage of cell subtypes in the sample is approximately 1.67%. In some cases, the percentage of cell subtypes in the sample is approximately 1%. In some cases, the percentage of cell subtypes in the sample is approximately 0%.

[0317] The reference doses or levels of macrophage biomarkers described herein may be based on the amount or level of macrophage biomarkers in a reference population. In some examples, the reference macrophage biomarkers described herein are the amount or level of macrophage biomarkers in a reference population that includes one or more subsets (e.g., two or more, three or more, four or more, or five or more) of patients.

[0318] In some examples, the reference macrophage biomarker is the quantity or level of macrophage biomarkers in a reference population, the reference population comprising at least one subset of patients having lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)).

[0319] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population comprising at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have received one or more doses (e.g., at least one, two, three, four, five, six, seven, eight, nine, or ten or more doses) of either an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0320] In some cases, the reference macrophage biomarker is the quantity or level of macrophage biomarkers in a reference population, the reference population includes at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab) as monotherapy.

[0321] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population including at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) as a combination therapy (e.g., anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab) and additional therapeutic agents (e.g., anticancer therapy (e.g., cytotoxic agents, growth inhibitors, radiotherapy, anti-angiogenic agents, or combinations thereof), e.g., CHP).

[0322] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population including at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab) as part of a combination therapy (e.g., anti-CD20 antibody (e.g., obinutuzumab or rituximab) and additional therapeutic agents (e.g., anticancer therapy (e.g., cytotoxic agents, growth inhibitors, radiotherapy, anti-angiogenic agents, or combinations thereof), e.g., CHOP).

[0323] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population includes at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with anti-cancer therapies (e.g., cytotoxic agents, growth inhibitors, radiotherapy, anti-angiogenic agents, or combinations thereof), e.g., CHP, but without anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

[0324] In some examples, the reference macrophage biomarker is the amount or level of macrophage biomarkers in a reference population, the reference population includes at least one subset of patients with lymphoma (e.g., B-cell lymphoma, e.g., non-Hodgkin lymphoma, e.g., diffuse large B-cell lymphoma (e.g., germinal center B-cell-like or activated B-cell-like diffuse large B-cell lymphoma)) who have been treated with anti-cancer therapy (e.g., cytotoxic agents, growth inhibitors, radiotherapy, anti-angiogenic agents, or combinations thereof), e.g., CHOP, and who have not received anti-CD20 antibodies (e.g., obinutuzumab or rituximab).

[0325] For example, in some cases, the reference population includes a first subset of patients who have been treated with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), and a second subset of patients who have not been treated with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0326] For example, in some cases, the reference population includes a first subset of patients who have been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab) and a second subset of patients who have not been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0327] For example, in some cases, the reference population includes a first subset of patients who have been treated with an anti-CD79b immunoconjugate (e.g., polatuzumab vedotin) and an anti-CD20 antibody (e.g., obinutuzumab or rituximab), and a second subset of patients who have been treated with an anti-CD20 antibody (e.g., obinutuzumab or rituximab).

[0328] In some cases, baseline levels or doses of macrophage biomarkers significantly separate each of the first and second subsets of patients based on the significant difference between patient responsiveness (e.g., PFS or OS) to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) and patient responsiveness to treatment with anti-CD20 antibodies (e.g., obinutuzumab or rituximab) below baseline macrophage biomarker levels, and patient responsiveness to treatment with anti-CD79b immunoconjugates (e.g., polatuzumab vedotin) and anti-CD20 antibodies (e.g., obinutuzumab or rituximab) is significantly improved compared to patient responsiveness to treatment with anti-CD20 antibodies (e.g., obinutuzumab or rituximab). For...

Claims

1. A method for identifying, diagnosing and / or predicting whether a patient with diffuse large B-cell lymphoma (DLBCL) can benefit from a treatment comprising an immunoconjugate and an anti-CD20 antibody, comprising measuring a macrophage biomarker in a sample from the patient, wherein the amount or level of the macrophage biomarker in the sample is below a reference macrophage biomarker amount or level, thereby identifying, diagnosing and / or predicting the patient to benefit from the treatment comprising the immunoconjugate and the anti-CD20 antibody, wherein the immunoconjugate is of the following formula: 【Chemistry 1】 A method comprising (wherein Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

2. A method for selecting a therapy for a patient having DLBCL, comprising measuring a macrophage biomarker in a sample from the patient, wherein the amount or level of the macrophage biomarker in the sample is below a reference amount or level of macrophage biomarker, thereby identifying the patient as being able to benefit from a treatment comprising an immunoconjugate and an anti-CD20 antibody, wherein the immunoconjugate is of the following formula: 【Chemistry 2】 A method comprising (wherein Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

3. The method according to claim 1 or 2, wherein the amount or level of the macrophage biomarker from the patient is below the amount or level of the reference macrophage biomarker, and the method further comprises administering an effective amount of the immunoconjugate and an effective amount of the anti-CD20 antibody to the patient.

4. A method for treating a patient with DLBCL, (a) Measuring macrophage biomarkers in a sample from the patient, wherein the amount or level of macrophage biomarkers in the sample is below the amount or level of reference macrophage biomarkers, and (b) Based on the macrophage biomarker measured in step (a), administer to the patient an effective amount of immunoconjugate and an effective amount of anti-CD20 antibody, wherein the immunoconjugate has the following formula: 【Transformation 3】 A method comprising (wherein Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

5. A method for treating a patient having DLBCL, comprising administering to the patient an effective amount of an immunoconjugate and an effective amount of anti-CD20 antibody, wherein, prior to treatment, it has been determined that the amount or level of a macrophage biomarker in a sample from the patient is below the amount or level of a reference macrophage biomarker, and the immunoconjugate has the following formula: 【Chemistry 4】 A method comprising (wherein Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

6. A method for treating a patient having DLBCL and in which the amount or level of macrophage biomarkers in a patient sample is below the reference amount or level of macrophage biomarkers, comprising administering to the patient an effective amount of an immunoconjugate and an effective amount of anti-CD20 antibody, wherein the immunoconjugate has the following formula: 【Transformation 5】 A method comprising (wherein Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8).

7. The method according to any one of claims 1 to 6, wherein the patient is a human patient.

8. The method according to any one of claims 1 to 7, wherein the reference macrophage biomarker amount or level is a pre-assigned macrophage biomarker amount or level.

9. The method according to any one of claims 1 to 8, wherein the reference macrophage biomarker amount or level is the amount or level of macrophage biomarker in a reference population.

10. The method according to claim 9, wherein the amount or level of the macrophage biomarker in the reference population is the median amount or level of the macrophage biomarker in the reference population.

11. The method according to claim 9, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 25th percentile of the reference population.

12. The method according to claim 9, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 50th percentile of the reference population.

13. The method according to claim 9, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 75th percentile of the reference population.

14. The method according to any one of claims 9 to 13, wherein the reference population is a population of patients having DLBCL.

15. The method according to claim 14, wherein the population of patients having DLBCL has been previously treated with the immunoconjugate and the anti-CD20 antibody.

16. The method according to claim 14, wherein the population of patients having DLBCL has been previously treated with the anti-CD20 antibody.

17. The method according to claim 14, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker in the reference population before initiation of treatment with the immunoconjugate and the anti-CD20 antibody.

18. The method according to any one of claims 1 to 17, wherein the reference macrophage biomarker amount or level is the amount of macrophages measured by gene expression.

19. The method according to claim 18, wherein the amount of macrophages is between approximately 0% and approximately 56.5%.

20. The method according to any one of claims 1 to 3, wherein the benefit is an extension of progression-free survival (PFS).

21. The method according to any one of claims 1 to 3, wherein the benefit is an increase in overall survival (OS).

22. The method according to any one of claims 4 to 6, further comprising achieving an improvement in PFS or OS.

23. The method according to any one of claims 1 to 22, wherein the macrophage biomarker is the average of the M1 macrophage gene signature set scores of one or more M1 macrophage gene signature sets.

24. The method according to claim 23, wherein each M1 macrophage gene signature set score is the average of the expression levels of one or more genes in the M1 macrophage gene signature set.

25. The method according to claim 24, wherein each M1 macrophage gene signature set score is the average of the normalized expression levels of one or more genes in the M1 macrophage gene signature set.

26. The aforementioned one or more M1 macrophage gene signature sets, (a) ACP2, ABCD1, C1QA, FDX1, CCL22, CD163, SCAMP2, ADAMDEC1, ARL8B, and HAMP, (b) ACP2, ABCD1, FDX1, CCL8, CCL22, CD163, ADAMDEC1, TREM2, and HAMP, (c) ACP2, ADRA2B, ALCAM, ABCD1, ATOX1, ATP6V0C, ATP6V1E1, BLVRA, C1QA, CD48, CD63, CLCN7, TPP1, CLTC, CCR1, CMKLR1, SLC31A1, COX5B, FCER1G, FDX1, FOLR2, FPR3, FTL, HEXB, HK3, IL10, IL12B, ITGAE, LAIR1, CXCL9, MMP19, NARS, NDUFS2, P2RX7, PDCL, MAPK13, PTGIR, PTPRA, RELA, CCL7, CCL8, CCL19, CCL22, SRC, STX4, TCEB1, TFRC, AGPS, MARCO, SNX3, CD84, USP14, ITGB1BP1, ATP6V1F, TRIP4, CD163, CIAO1, WTAP, ARHGEF11, ABI1, SCAMP2, ACTR2, BCAP31, ZMPSTE24, BCKDK, EXOC5, STIP1, UQCR11, SDS, LILRB4, OGF, TFEC, FKBP15, DNAJC13, TDRD7, STX12, IL17RA, ABTB2, FAM32A, SIGLEC7, SIGLEC9, ADAMDEC1, CECR5, SLC25A24, NRBP1, MS4A4A, TREM2, OTUD4, PQLC2, HAUS2, ARL8B, NECAP2, WDR11, ZC3H15, CCDDC47, UTP3, MRS2, HAMP, MRPL40, VPS33A, CORO7, LIMD2, TMX1, DOT1L, ADO, and ADCK2, (d) ACP2, ADRA2B, ALCAM, TSPO, C3AR1, DAGLA, CALR, CHIT1, CYBB, CYC1, CYP19A1, DLAT, FCER1G, GP1BA, GPD1, IFNAR1, IL10, KCNJ5, KIFC3, MT2A, MYBPH, MYH11, MYO7A, P2RX7, PRDX1, RAB3IL1, RNH1, MRPL12, CCL1, CCL7, CCL8, CCL24, SRC, VIM, RRP1, MARCO, S1PR2, AP1M2, ACTR3, LILRB1, AFG3L2, SDS, LILRB4, EMILIN1, VSIG4, HSPB7, COQ2, ADAMDEC1, CECR5, WSB2, SLAMF8, DNASE2B, CLPB, MFSD7, and ADCK2, (SAM2、SAMA33、SAMA20、SAMA2、SAS1 FAMOUS、FAMOUS、FAMOUS、FAMOUS33 C1、SYS11、SYS1、SYS3、SYS1 1101、CHY10、CHY1、CHY3、CHY 100001000000000000000000000000000000 33、HAR10、HAR10、HYS1、DY20、DYS 20207、00000000000000000000000000000000000000000000000. ROSE7、ROSE8、ROSE18、ROSE19、ROSE24、ROSE1 、SAKA、SAX、SAS1、SASHA、SHA201、SAS22、S 4. 163、LOVE1、DY102、CHSIS、LOVES10、SHR、4 LOVE 4、 LOVE1 1 LOVE1 CHEEK1、HARSH15、HARSHY、HARHYH、HARSHAR4、HARSH 11、DYS2、DYSYS1、DYSYS1、DYSYS 10、SHAS10、SHAS2、SHAS20、SHAS70、SHAS C2、DY33、SYSY8、SYSY、DYSYSY、F1 11. LOVE3, LOVE4, and LOVE2 (f) ACP2, ADCY3, ADRA2B, ALCAM, ABCD1, ANXA2, ATP6V1A, C1QA, C1QB, C3AR1, DAGLA, CD80, CD63, CHIT1, CMKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP19A 1, FANCE, FDX1, FPR2, FPR3, GPD1, HEXB, KCNJ1, KCNJ5, KIFC3, MMP19, MSR1, M T2A, MYBPH, P2RX7, MAPK13, S100A11, CCL1, CCL7, CCL8, CCL18, CCL19, CCL22, CCL24, SLC1A2, SLC6A12, SLC11A1, SIGLEC1, SRC, TIE1, MARCO, HYAL2, CD163 , LONP1, IGSF6, LILRB1, CD300C, SDS, LILRB4, EMILIN1, VSIG4, PHLDB1, NCAPH , CLEC4E, MYOF, HSPB7, ADAMDEC1, GLRX2, MS4A4A, ATP6V1H, TREM2, TMEM70, T MEM33, KCNK13, SLAMF8, HAMP, DNASE2B, MYOZ1, MFSD7, ADO, ADCK2, and TBC1D16 The method according to claim 24 or 25.

27. The method according to any one of claims 1 to 22, wherein the macrophage biomarker is the average of the TAM gene signature set scores of one or more TAM gene signature sets.

28. The method according to claim 27, wherein each TAM gene signature set score is the average of the expression levels of one or more genes in the TAM gene signature set.

29. The method according to claim 28, wherein each TAM gene signature set score is the average of the normalized expression levels of one or more genes in the TAM gene signature set.

30. The one or more TAM gene signature sets described above, (a) MARCO, ACP5, VSIG4, MRC1, MSR1, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, C1QC, APOE, FOLR2, CTSD and SPP1 The method according to claim 28 or 29.

31. The method according to any one of claims 1 to 22, wherein the macrophage biomarker is a gene expression value.

32. The method according to claim 31, wherein the gene expression value is the median of the gene expression values.

33. The method according to claim 31 or 32, wherein the gene expression value is measured using a gene signature matrix.

34. The aforementioned gene signature matrix includes the following genes: (a) CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SEMAG3, CD8A, TOGARAM2, COLALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, HOPX, CD1D, GNG7, TCF4, BANK1, FHIT, FCMR, GNG2, GFRRA2, KBTBD11, RALGPS2, TSPOAP1, PLEKH1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRRC32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVR1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PTGS1, TMEM170B, TREMB2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQR, INHBA, and ICAM1, or (b)CD200、KLHL14、TCL1A、NRG1、CYP4 F3、EOMES、PPP2R2B、RNF165、WNT7A、C CR4、PDGFD、EBF1、FCGBP、PCDH9、MLC1 、TSHZ2、S1PR5、NCALD、LAYN、CD248、GC NT4、FASLG、TRAT1、ADAM6、GUCY1A3、L RRC4、TSPAN18、SBK1、ICOS、BTNL8、WN T5B、AUTS2、SH2D2A、ADGRG3、PNOC、SPIB、VPREB3、DPEP3、MME、ZBTB16、FOXP3 、SEMA3G、CD8A、TOGARAM2、COLGALT2、 ABCB1、STAP1、SAMD3、FAM46C、BLK、CT LA4、CD19、REPS2、RTKN2、POU2AF1、DA PK2、PYHIN1、NLRC3、GATM、KLRD1、AFF3 、FCRLA、AATBC、REM2、YPEL1、TXK、CD8 B、P2RX5、CEACAM1、BCL11A、NINJ2、AB CB4、CD5、HAL、HPGD、BLNK、PLCL1、CEP 19、HPSE、SLFN13、HOPX、CD1D、GNG7、TM EM154、TCF4、BANK1、FHIT、FCMR、GNG2 、GFRA2、KBTBD11、TECPR2、RALGPS2、T SPOAP1、PLEKHF1、MEF2C、MAOA、TTYH2 、HLA-DOB、NRGN、DGAT2、FXYD6、TMCC3、 MGAM、TTC38、LRRC32、ARHGAP24、PPP1 R3B、STAT4、SLC7A8、CD72、FZD1、GK5、 DYSF、PLTP、SMARCD3、FAM160B1、PDPN 、AKAP2、ACVRL1、KCNJ15、CD36、ALDH1A 2、ENPP2、COLEC12、PTGS1、TMEM170B、 DOCK5、TREM2、C5AR2、ECM1、SLC1A3、A BHD5、MS4A4A、CLIC2、IL1R1、SLC2A6、 GAS7、RNF144B、SLC6A12、FPR2、ADAM28 、GRK3、KDM1B、MATK、LMO2、CFB、CCRL2 、CLEC4A、TLR4、LILRA2、ACE、TLR1、LRR K2、LY96、NUPR1、CISH、CSTA、EREG、AD AMDEC1、RNASE6、CXCL3、VSIG4、CXCL2、CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1, The method according to claim 33, comprising

35. The gene signature matrix is ​​the following genes: CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, M ME, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAM D3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLR C3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEAC AM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, HOPX, CD1D, GNG 7, TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLE KHF1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LR RC32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FA M160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PTGS1, TM The method according to claim 34, comprising EM170B, TREM2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1.

36. The method according to any one of claims 33 to 35, wherein the gene signature matrix is ​​used to determine the number of M1 macrophages or tumor-associated macrophages.

37. The method according to any one of claims 1 to 22, wherein the macrophage biomarker is the amount of M1 macrophages or the amount of tumor-associated macrophages.

38. The method according to claim 37, wherein the amount of M1 macrophages or tumor-associated macrophages is measured directly or indirectly.

39. The method according to claim 38, wherein the amount of M1 macrophages or tumor-associated macrophages is directly measured using flow cytometry, spatial transcriptomics, spatial proteomics, or a combination thereof.

40. The method according to claim 38, wherein the amount of M1 macrophages or tumor-associated macrophages is measured indirectly using nucleic acids or proteins.

41. The method according to claim 40, wherein the nucleic acid is measured using RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

42. The method according to claim 41, wherein the amount of M1 macrophages or tumor-associated macrophages is measured using a marker gene approach or a deconvolution approach.

43. The method according to claim 42, wherein the marker gene approach uses xCell.

44. The method according to claim 42, wherein the deconvolution approach uses quantiseq.

45. The method according to any one of claims 1 to 22, wherein the macrophage biomarker in the sample from the patient is measured using nucleic acid or protein.

46. The method according to claim 45, wherein the macrophage biomarker in the sample from the patient is determined using nucleic acid expression levels.

47. The method according to claim 46, wherein the nucleic acid expression level is determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

48. The method according to claim 46 or 47, wherein the nucleic acid expression level is the mRNA expression level.

49. The method according to claim 48, wherein the mRNA expression level is determined by RNA-seq.

50. The method according to any one of claims 1 to 49, wherein the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

51. The method according to claim 50, wherein the sample is a tissue sample.

52. The method according to claim 51, wherein the tissue sample is a tumor tissue sample.

53. The method according to claim 52, wherein the tumor tissue sample comprises tumor cells, tumor-infiltrating immune cells, stromal cells, normal adjacent tissue (NAT) cells, or a combination thereof.

54. The method according to claim 52 or 53, wherein the tumor tissue sample is a biopsy material.

55. The method according to any one of claims 50 to 54, wherein the sample is an archived sample, a fresh sample, or a frozen sample.

56. The method according to any one of claims 1 to 55, wherein the DLBCL is a subgroup of germinal center B cell-like (GCB) or activated B cell-like (ABC) originating cells of DLBCL.

57. The method according to any one of claims 1 to 56, wherein the DLBCL is CD79b-positive DLBCL and / or CD20-positive DLBCL.

58. The method according to any one of claims 1 to 57, wherein the patient has not been previously treated for DLBCL.

59. The method according to any one of claims 1 to 58, wherein the patient has not been previously administered the immunoconjugate and the anti-CD20 antibody.

60. The method according to any one of claims 1 to 59, wherein the anti-CD79b antibody comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) containing the amino acid sequence of SEQ ID NO:

4.

61. The aforementioned anti-CD79b antibody, (a) A heavy chain containing the amino acid sequence of SEQ ID NO: 13 and a light chain containing the amino acid sequence of SEQ ID NO: 11, or (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 14, or (c) Heavy chain containing the amino acid sequence of SEQ ID NO: 12 and light chain containing the amino acid sequence of SEQ ID NO: 11 The method according to any one of claims 1 to 60, including the method described in any one of claims 1 to 60.

62. The method according to any one of claims 1 to 61, wherein p is 2 to 7, 2 to 6, 2 to 5, 3 to 5, or 3 to 4.

63. The method according to claim 62, wherein p is 3.

4.

64. The method according to claim 62, wherein p is 3.

5.

65. The method according to any one of claims 1 to 64, wherein the immunoconjugate is polatuzumab vedotin.

66. The method according to any one of claims 1 to 65, wherein the anti-CD20 antibody is a type I anti-CD20 antibody or a type II anti-CD20 antibody.

67. The method according to claim 66, wherein the anti-CD20 antibody is a type I anti-CD20 antibody.

68. The aforementioned type I anti-CD20 antibody is the following CDR: (a) CDR-H1 having the amino acid sequence of SEQ ID NO: 26, (b) CDR-H2 having the amino acid sequence of SEQ ID NO: 27, (c) CDR-H3 having the amino acid sequence of SEQ ID NO: 28, (d) CDR-L1 having the amino acid sequence of SEQ ID NO: 29, (e) CDR-L2 having the amino acid sequence of SEQ ID NO: 30, (f) CDR-L3 having the amino acid sequence of SEQ ID NO: 31 The method according to claim 67, including the method described in claim 67.

69. The method according to claim 68, wherein the type I anti-CD20 antibody comprises a VH domain containing the amino acid sequence of SEQ ID NO: 40 and a VL domain containing the amino acid sequence of SEQ ID NO:

41.

70. The method according to claim 69, wherein the type I anti-CD20 antibody is rituximab.

71. The method according to any one of claims 65 to 70, wherein polatuzumab vedotin is administered in a dose of about 1.0 mg / kg to about 1.8 mg / kg.

72. The method according to claim 71, wherein polatuzumab vedotin is administered at a dose of approximately 1.8 mg / kg.

73. Rituximab, 375 mg / m² ２ The method according to any one of claims 70 to 72, administered in the dose of [specified dose].

74. The method according to any one of claims 65 to 73, wherein polatuzumab vedotin and / or rituximab is administered intravenously.

75. The method according to any one of claims 3 to 74, further comprising administering an effective amount of an additional therapeutic agent to the patient.

76. The method according to claim 75, wherein the additional therapeutic agent is one or more of the following: chemotherapeutic agents, corticosteroids, antineoplastic agents, growth inhibitors, anti-angiogenic agents, radiotherapy, cytotoxic agents, or a combination thereof.

77. The method according to claim 76, wherein the additional therapeutic agent is a chemotherapeutic agent and a corticosteroid.

78. The method according to claim 76 or 77, wherein the chemotherapeutic agent is cyclophosphamide and / or doxorubicin.

79. The method according to any one of claims 76 to 78, wherein the corticosteroid is prednisone, prednisolone, or methylprednisolone.

80. Cyclophosphamide at approximately 375 mg / m² ２ ~Approx. 750mg / m ２ The method according to claim 78 or 79, administered in the dose of [specify dose].

81. Doxorubicin at approximately 25 mg / m² ２ ~Approx. 50mg / m ２ The method according to any one of claims 78 to 80, administered in the dose of [specified dose].

82. (a) Prednisone is administered at a dose of approximately 100 mg, or (b) Prednisolone is administered in a dose of approximately 100 mg, or (c) Methylprednisolone is administered at a dose of approximately 80 mg. The method according to any one of claims 79 to 81.

83. The method according to any one of claims 78 to 82, wherein cyclophosphamide and / or doxorubicin are administered intravenously.

84. The method according to any one of claims 79 to 83, wherein prednisone, prednisolone, or methylprednisolone is administered orally.

85. The method according to any one of claims 79 to 84, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin and / or prednisone, prednisolone or methylprednisolone is administered in at least one 21-day cycle.

86. (a) Polatuzumab vedotin, rituximab, cyclophosphamide and / or doxorubicin are administered on day 1 of each 21-day cycle, and / or (b) Prednisone, prednisolone, or methylprednisolone is administered on days 1 to 5 of each 21-day cycle. The method according to claim 85.

87. The method according to any one of claims 85 to 86, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone, prednisolone, or methylprednisolone is administered in one, two, three, four, five, or six 21-day cycles.

88. The use of an immunoconjugate and an anti-CD20 antibody in the manufacture of a pharmaceutical for the treatment of DLBCL, for treating patients in whom the amount or level of macrophage biomarkers in a patient sample is below the reference amount or level of macrophage biomarkers, wherein the immunoconjugate is of the following formula: 【Transformation 6】 (In the formula, Ab is an anti-CD79b antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8) used.

89. The use according to claim 88, wherein the patient is a human patient.

90. The use according to claim 88 or 89, wherein the reference macrophage biomarker amount or level is a pre-assigned macrophage biomarker amount or level.

91. The use according to any one of claims 88 to 90, wherein the reference macrophage biomarker amount or level is the amount or level of macrophage biomarker in a reference population.

92. The use according to claim 91, wherein the amount or level of the macrophage biomarker in the reference population is the median amount or level of the macrophage biomarker in the reference population.

93. The use according to claim 91, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 25th percentile of the reference population.

94. The use according to claim 91, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 50th percentile of the reference population.

95. The use according to claim 91, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 75th percentile of the reference population.

96. The use according to any one of claims 91 to 95, wherein the reference population is a population of patients having DLBCL.

97. The use according to claim 96, wherein the population of patients having DLBCL has been previously treated with the immunoconjugate and the anti-CD20 antibody.

98. The use according to claim 96, wherein the population of patients having DLBCL has been previously treated with the anti-CD20 antibody.

99. The use according to claim 96, wherein the reference macrophage biomarker amount or level is the amount or level of macrophage biomarker in the reference population before initiating treatment with the immunoconjugate and the anti-CD20 antibody.

100. The use according to any one of claims 88 to 99, wherein the reference macrophage biomarker amount or level is the amount of macrophages measured by gene expression.

101. The use according to claim 100, wherein the amount of macrophages is between approximately 0% and approximately 56.5%.

102. The use according to any one of claims 88 to 101, wherein the treatment achieves improvement in PFS or OS.

103. The use according to any one of claims 88 to 102, wherein the macrophage biomarker is the average of the M1 macrophage gene signature set scores of one or more M1 macrophage gene signature sets.

104. The use according to claim 103, wherein each M1 macrophage gene signature set score is the average of the expression levels of one or more genes in the M1 macrophage gene signature set.

105. The use according to claim 104, wherein each M1 macrophage gene signature set score is the mean of the normalized expression levels of one or more genes in the M1 macrophage gene signature set.

106. The aforementioned one or more M1 macrophage gene signature sets, (a) ACP2, ABCD1, C1QA, FDX1, CCL22, CD163, SCAMP2, ADAMDEC1, ARL8B, and HAMP, (b) ACP2, ABCD1, FDX1, CCL8, CCL22, CD163, ADAMDEC1, TREM2, and HAMP, (c) ACP2, ADRA2B, ALCAM, ABCD1, ATOX1, ATP6V0C, ATP6V1E1, BLVRA, C1QA, CD48, CD63, CLCN7, TPP1, CLTC, CCR1, CMKLR1, SLC31A1, COX5B, FCER1G, FDX1, FOLR2, FPR3, FTL, HEXB, HK3, IL10, IL12B, ITGAE, LAIR1, CXCL9, MMP19, NARS, NDUFS2, P2RX7, PDCL, MAPK13, PTGIR, PTPRA, RELA, CCL7, CCL8, CCL19, CCL22, SRC, STX4, TCEB1, TFRC, AGPS, MARCO, SNX3, CD84, USP14, ITGB1BP1, ATP6V1F, TRIP4, CD163, CIAO1, WTAP, ARHGEF11, ABI1, SCAMP2, ACTR2, BCAP31, ZMPSTE24, BCKDK, EXOC5, STIP1, UQCR11, SDS, LILRB4, OGF, TFE, FKBP15, DNAJC13, TDRD7, STX12, IL17RA, ABTB2, FAM32A, SIGLEC7, SIGLEC9, ADAMDEC1, CECR5, SLC25A24, NRBP1, MS4A4A, TREM2, OTUD4, PQLC2, HAUS2, ARL8B, NECAP2, WDR11, ZC3H15, CCD47, UTP3, MRS2, HAMP, MRPL40, VPS33A, CORO7, LIMD2, TMX1, DOT1L, ADO, and ADCK2, (d) ACP2, ADRA2B, ALCAM, TSPO, C3AR1, DAGLA, CALR, CHIT1, CYBB, CYC1, CYP19A1, DLAT, FCER1G, GP1BA, GPD1, IFNAR1, IL10, KCNJ5, KIFC3, MT2A, MYBPH, MYH11, MYO7A, P2RX7, PRDX1, RAB3IL1, RNH1, MRPL12, CCL1, CCL7, CCL8, CCL24, SRC, VIM, RRP1, MARCO, S1PR2, AP1M2, ACTR3, LILRB1, AFG3L2, SDS, LILRB4, EMILIN1, VSIG4, HSPB7, COQ2, ADAMDEC1, CECR5, WSB2, SLAMF8, DNASE2B, CLPB, MFSD7, and ADCK2, (SAM2、SAMA33、SAMA20、SAMA2、SAS1 FAMOUS、FAMOUS、FAMOUS、FAMOUS33 C1、SYS11、SYS1、SYS3、SYS1 1101、CHY10、CHY1、CHY3、CHY 100001000000000000000000000000000000 33、HAR10、HAR10、HYS1、DY20、DYS 20207、00000000000000000000000000000000000000000000000. ROSE7、ROSE8、ROSE18、ROSE19、ROSE24、ROSE1 、SAKA、SAX、SAS1、SASHA、SHA201、SAS22、S 4. 163、LOVE1、DY102、CHSIS、LOVES10、SHR、4 LOVE 4、 LOVE1 1 LOVE1 CHEEK1、HARSH15、HARSHY、HARHYH、HARSHAR4、HARSH 11、DYS2、DYSYS1、DYSYS1、DYSYS 10、SHAS10、SHAS2、SHAS20、SHAS70、SHAS C2、DY33、SYSY8、SYSY、DYSYSY、F1 11. LOVE3, LOVE4, and LOVE2 (f) ACP2, ADCY3, ADRA2B, ALCAM, ABCD1, ANXA2, ATP6V1A, C1QA, C1QB, C3AR1, DAGLA, CD80, CD63, CHIT1, CMKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP19A 1, FANCE, FDX1, FPR2, FPR3, GPD1, HEXB, KCNJ1, KCNJ5, KIFC3, MMP19, MSR1, M T2A, MYBPH, P2RX7, MAPK13, S100A11, CCL1, CCL7, CCL8, CCL18, CCL19, CCL22, CCL24, SLC1A2, SLC6A12, SLC11A1, SIGLEC1, SRC, TIE1, MARCO, HYAL2, CD163 , LONP1, IGSF6, LILRB1, CD300C, SDS, LILRB4, EMILIN1, VSIG4, PHLDB1, NCAPH , CLEC4E, MYOF, HSPB7, ADAMDEC1, GLRX2, MS4A4A, ATP6V1H, TREM2, TMEM70, T MEM33, KCNK13, SLAMF8, HAMP, DNASE2B, MYOZ1, MFSD7, ADO, ADCK2, and TBC1D16 The use according to claim 104 or 105.

107. The use according to any one of claims 88 to 102, wherein the macrophage biomarker is the average of the tumor-associated macrophage gene signature set scores of one or more tumor-associated macrophage gene signature sets.

108. The use according to claim 107, wherein each tumor-associated macrophage gene signature set score is the average of the expression levels of one or more genes in the tumor-associated macrophage gene signature set.

109. The use according to claim 108, wherein each tumor-associated macrophage gene signature set score is the mean of the normalized expression levels of one or more genes in the tumor-associated macrophage gene signature set.

110. The aforementioned set of one or more tumor-associated macrophage gene signatures (g) MARCO, ACP5, VSIG4, MRC1, MSR1, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, C1QC, APOE, FOLR2, CTSD and SPP1 The use according to claim 107 or 108.

111. The use according to any one of claims 88 to 102, wherein the macrophage biomarker is a gene expression value.

112. The use according to claim 107, wherein the gene expression value is the median of the gene expression values.

113. The use according to claim 107 or 112, wherein the gene expression value is measured using a gene signature matrix.

114. The aforementioned gene signature matrix includes the following genes: (a) CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOXP3, SEMAG3, CD8A, TOGARAM2, COLALT2, ABCB1, STAP1, SAMD3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLC1, HPSE, SLFN13, HOPX, CD1D, GNG7, TCF4, BANK1, FHIT, FCMR, GNG2, GFR2, KBTBD11, RALGPS2, TSPOAP1, PLEKH1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRRC32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVR1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PTGS1, TMEM170B, TRE2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQR, INHBA, and ICAM1, or (b)CD200、KLHL14、TCL1A、NRG1、CYP4 F3、EOMES、PPP2R2B、RNF165、WNT7A、C CR4、PDGFD、EBF1、FCGBP、PCDH9、MLC1 、TSHZ2、S1PR5、NCALD、LAYN、CD248、GC NT4、FASLG、TRAT1、ADAM6、GUCY1A3、L RRC4、TSPAN18、SBK1、ICOS、BTNL8、WN T5B、AUTS2、SH2D2A、ADGRG3、PNOC、SPIB、VPREB3、DPEP3、MME、ZBTB16、FOXP3 、SEMA3G、CD8A、TOGARAM2、COLGALT2、 ABCB1、STAP1、SAMD3、FAM46C、BLK、CT LA4、CD19、REPS2、RTKN2、POU2AF1、DA PK2、PYHIN1、NLRC3、GATM、KLRD1、AFF3 、FCRLA、AATBC、REM2、YPEL1、TXK、CD8 B、P2RX5、CEACAM1、BCL11A、NINJ2、AB CB4、CD5、HAL、HPGD、BLNK、PLCL1、CEP 19、HPSE、SLFN13、HOPX、CD1D、GNG7、TM EM154、TCF4、BANK1、FHIT、FCMR、GNG2 、GFRA2、KBTBD11、TECPR2、RALGPS2、T SPOAP1、PLEKHF1、MEF2C、MAOA、TTYH2 、HLA-DOB、NRGN、DGAT2、FXYD6、TMCC3、 MGAM、TTC38、LRRC32、ARHGAP24、PPP1 R3B、STAT4、SLC7A8、CD72、FZD1、GK5、 DYSF、PLTP、SMARCD3、FAM160B1、PDPN 、AKAP2、ACVRL1、KCNJ15、CD36、ALDH1A 2、ENPP2、COLEC12、PTGS1、TMEM170B、 DOCK5、TREM2、C5AR2、ECM1、SLC1A3、A BHD5、MS4A4A、CLIC2、IL1R1、SLC2A6、 GAS7、RNF144B、SLC6A12、FPR2、ADAM28 、GRK3、KDM1B、MATK、LMO2、CFB、CCRL2 、CLEC4A、TLR4、LILRA2、ACE、TLR1、LRR K2、LY96、NUPR1、CISH、CSTA、EREG、AD AMDEC1、RNASE6、CXCL3、VSIG4、CXCL2、CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1, The use according to claim 113, comprising

115. The gene signature matrix includes the following genes: CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MM E, ZBTB16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD 3, FAM46C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC 3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACA M1, BCL11A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, HOPX, CD1D, GNG7 , TCF4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLEK HF1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRR C32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM 160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PTGS1, TME The use according to claim 114, comprising M170B, TREM2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1.

116. The use according to any one of claims 113 to 115, wherein the gene signature matrix is ​​used to determine the number of M1 macrophages or tumor-associated macrophages.

117. The use according to any one of claims 88 to 102, wherein the macrophage biomarker is the amount of M1 macrophages or the amount of tumor-associated macrophages.

118. The use according to claim 117, wherein the amount of M1 macrophages or tumor-associated macrophages is measured directly or indirectly.

119. The use according to claim 118, wherein the amount of M1 macrophages or tumor-associated macrophages is measured directly using flow cytometry, spatial transcriptomics, spatial proteomics, or a combination thereof.

120. The use according to claim 118, wherein the amount of M1 macrophages or tumor-associated macrophages is measured indirectly using nucleic acids or proteins.

121. The use according to claim 120, wherein the nucleic acid is measured using RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

122. The use according to claim 121, wherein the amount of M1 macrophages or tumor-associated macrophages is measured using a marker gene approach or a deconvolution approach.

123. The use according to claim 122, wherein the marker gene approach uses xCell.

124. The use according to claim 122, wherein the deconvolution approach uses quantIseq.

125. The use according to any one of claims 88 to 102, wherein the macrophage biomarker in the sample from the patient is measured using nucleic acid or protein.

126. The use according to claim 125, wherein the macrophage biomarker in the sample from the patient is determined using nucleic acid expression levels.

127. The use according to claim 126, wherein the nucleic acid expression level is determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

128. The use according to claim 126 or 127, wherein the nucleic acid expression level is the mRNA expression level.

129. The use according to claim 128, wherein the mRNA expression level is determined by RNA-seq.

130. The use according to any one of claims 88 to 129, wherein the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

131. The use according to claim 130, wherein the sample is a tissue sample.

132. The use according to claim 131, wherein the tissue sample is a tumor tissue sample.

133. The use according to claim 132, wherein the tumor tissue sample comprises tumor cells, tumor-infiltrating immune cells, stromal cells, normal adjacent tissue (NAT) cells, or a combination thereof.

134. The use according to claim 132 or 133, wherein the tumor tissue sample is a biopsy material.

135. The use according to any one of claims 130 to 134, wherein the sample is an archived sample, a fresh sample, or a frozen sample.

136. The use according to any one of claims 88 to 135, wherein the DLBCL is a subgroup of germinal center B cell-like (GCB) or activated B cell-like (ABC) originating cells of DLBCL.

137. The use according to any one of claims 88 to 136, wherein the DLBCL is CD79a-positive DLBCL and / or CD20-positive DLBCL.

138. The use according to any one of claims 88 to 137, wherein the patient has not been previously treated for DLBCL.

139. The use according to any one of claims 88 to 138, wherein the patient has not been previously administered the immunoconjugate and the anti-CD20 antibody.

140. The use according to any one of claims 88 to 139, wherein the anti-CD79b antibody comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) containing the amino acid sequence of SEQ ID NO:

4.

141. The aforementioned anti-CD79b antibody, (a) A heavy chain containing the amino acid sequence of SEQ ID NO: 13 and a light chain containing the amino acid sequence of SEQ ID NO: 11, or (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 14, or (c) Heavy chain containing the amino acid sequence of SEQ ID NO: 12 and light chain containing the amino acid sequence of SEQ ID NO: 11 Use according to any one of claims 88 to 140, including the use described in any one of claims 88 to 140.

142. The use according to any one of claims 88 to 141, wherein p is 2 to 7, 2 to 6, 2 to 5, 3 to 5, or 3 to 4.

143. The use according to claim 142, wherein p is 3.

4.

144. The use according to claim 142, wherein p is 3.

5.

145. The use according to any one of claims 88 to 144, wherein the immunoconjugate is polatuzumab vedotin.

146. The use according to any one of claims 88 to 145, wherein the anti-CD20 antibody is a type I anti-CD20 antibody or a type II anti-CD20 antibody.

147. The use according to claim 146, wherein the anti-CD20 antibody is a type I anti-CD20 antibody.

148. The aforementioned type I anti-CD20 antibody is the following CDR: (a) CDR-H1 having the amino acid sequence of SEQ ID NO: 26, (b) CDR-H2 having the amino acid sequence of SEQ ID NO: 27, (c) CDR-H3 having the amino acid sequence of SEQ ID NO: 28, (d) CDR-L1 having the amino acid sequence of SEQ ID NO: 29, (e) CDR-L2 having the amino acid sequence of SEQ ID NO: 30, (f) CDR-L3 having the amino acid sequence of SEQ ID NO: 31 The use according to claim 147, including the use described in claim 147.

149. The use according to claim 148, wherein the type I anti-CD20 antibody comprises a VH domain containing the amino acid sequence of SEQ ID NO: 40 and a VL domain containing the amino acid sequence of SEQ ID NO:

41.

150. The use according to claim 149, wherein the type I anti-CD20 antibody is rituximab.

151. The use according to any one of claims 145 to 150, wherein polatuzumab vedotin is administered in a dose of about 1.0 mg / kg to about 1.8 mg / kg.

152. The use according to claim 151, wherein polatuzumab vedotin is administered at a dose of approximately 1.8 mg / kg.

153. Rituximab is administered at approximately 375 mg / m². ２ The use according to any one of claims 150 to 152, administered in the specified dose.

154. The use according to any one of claims 145 to 153, wherein polatuzumab vedotin and / or rituximab is administered intravenously.

155. The use according to any one of claims 88 to 154, wherein the pharmaceutical is administered to the patient in combination with an effective amount of an additional therapeutic agent.

156. The use according to claim 155, wherein the additional therapeutic agent is one or more of the following: chemotherapeutic agents, corticosteroids, antineoplastic agents, growth inhibitors, anti-angiogenic agents, radiotherapy, cytotoxic agents, or a combination thereof.

157. The use according to claim 156, wherein the additional therapeutic agent is a chemotherapeutic agent and a corticosteroid.

158. The use according to claim 156 or 157, wherein the chemotherapeutic agent is cyclophosphamide and / or doxorubicin.

159. The use according to any one of claims 156 to 158, wherein the corticosteroid is prednisone, prednisolone, or methylprednisolone.

160. Cyclophosphamide at approximately 375 mg / m² ２ ~Approx. 750mg / m ２ The use according to claim 158 or 159, administered in the specified dose.

161. Doxorubicin is administered at a dose of about 25 mg / m ２ to about 50 mg / m ２ The use according to any one of claims 158 to 160, wherein the use is administered at a dose of

162. (a) Prednisone is administered at a dose of approximately 100 mg, or (b) Prednisolone is administered in a dose of approximately 100 mg, or (c) Methylprednisolone is administered at a dose of approximately 80 mg. The use according to any one of claims 159 to 161.

163. The use according to any one of claims 158 to 162, wherein cyclophosphamide and / or doxorubicin are administered intravenously.

164. The use according to any one of claims 159 to 163, wherein prednisone, prednisolone, or methylprednisolone is administered orally.

165. The use according to any one of claims 159 to 164, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin and / or prednisone, prednisolone or methylprednisolone is administered in at least one 21-day cycle.

166. (a) Polatuzumab vedotin, rituximab, cyclophosphamide and / or doxorubicin are administered on day 1 of each 21-day cycle, and / or (b) Prednisone, prednisolone, or methylprednisolone is administered on days 1 to 5 of each 21-day cycle. The use described in claim 165.

167. The use according to any one of claims 165 to 166, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone, prednisolone, or methylprednisolone is administered in one, two, three, four, five, or six 21-day cycles.

168. An immunoconjugate and an anti-CD20 antibody for use in the treatment of patients having DLBCL and in which the amount or level of macrophage biomarkers in patient samples is below the reference amount or level of macrophage biomarkers, wherein the immunoconjugate has the following formula: 【Transformation 7】 An immunoconjugate and an anti-CD20 antibody comprising (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 containing the amino acid sequence of SEQ ID NO: 6, (iii) HVR-H3 containing the amino acid sequence of SEQ ID NO: 7, (iv) HVR-L1 containing the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 containing the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 containing the amino acid sequence of SEQ ID NO: 10, and p is 1 to 8.

169. The immunoconjugate and anti-CD20 antibody for use according to claim 168, wherein the patient is a human patient.

170. The immunoconjugate and anti-CD20 antibody for use according to claim 168 or 169, wherein the reference macrophage biomarker amount or level is a pre-assigned macrophage biomarker amount or level.

171. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 170, wherein the reference macrophage biomarker amount or level is the amount or level of macrophage biomarker in a reference population.

172. The immunoconjugate and anti-CD20 antibody for use according to claim 171, wherein the amount or level of the macrophage biomarker in the reference population is the median amount or level of the macrophage biomarker in the reference population.

173. The immunoconjugate and anti-CD20 antibody for use according to claim 171, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 25th percentile of the reference population.

174. The immunoconjugate and anti-CD20 antibody for use according to claim 171, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 50th percentile of the reference population.

175. The immunoconjugate and anti-CD20 antibody for use according to claim 171, wherein the reference macrophage biomarker amount or level is the amount or level of the macrophage biomarker at the 75th percentile of the reference population.

176. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 171 to 175, wherein the reference population is a population of patients having DLBCL.

177. The immunoconjugate and anti-CD20 antibody for use according to claim 176, wherein the population of patients having DLBCL has been previously treated with the immunoconjugate and the anti-CD20 antibody.

178. The immunoconjugate and anti-CD20 antibody for use according to claim 176, wherein the population of patients having DLBCL has been previously treated with the anti-CD20 antibody.

179. The immunoconjugate and anti-CD20 antibody for use according to claim 176, wherein the reference macrophage biomarker amount or level is the amount or level of macrophage biomarker in the reference population before initiating treatment with the immunoconjugate and the anti-CD20 antibody.

180. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 179, wherein the reference macrophage biomarker amount or level is the amount of macrophages measured by gene expression.

181. The immunoconjugate and anti-CD20 antibody for use according to claim 180, wherein the amount of macrophages is approximately 0% to approximately 56.5%.

182. An immunoconjugate and an anti-CD20 antibody for use according to any one of claims 168 to 181, wherein the treatment achieves improvement in PFS or OS.

183. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 182, wherein the macrophage biomarker is the average of the M1 macrophage gene signature set scores of one or more M1 macrophage gene signature sets.

184. The immunoconjugate and anti-CD20 antibody for use according to claim 183, wherein each M1 macrophage gene signature set score is the average of the expression levels of one or more genes in the M1 macrophage gene signature set.

185. The immunoconjugate and anti-CD20 antibody for use according to claim 184, wherein each M1 macrophage gene signature set score is the mean of the normalized expression levels of one or more genes in the M1 macrophage gene signature set.

186. The aforementioned one or more M1 macrophage gene signature sets, (a) ACP2, ABCD1, C1QA, FDX1, CCL22, CD163, SCAMP2, ADAMDEC1, ARL8B, and HAMP, (b) ACP2, ABCD1, FDX1, CCL8, CCL22, CD163, ADAMDEC1, TREM2, and HAMP, (c) ACP2, ADRA2B, ALCAM, ABCD1, ATOX1, ATP6V0C, ATP6V1E1, BLVRA, C1QA, CD48, CD63, CLCN7, TPP1, CLTC, CCR1, CMKLR1, SLC31A1, COX5B, FCER1G, FDX1, FOLR2, FPR3, FTL, HEXB, HK3, IL10, IL12B, ITGAE, LAIR1, CXCL9, MMP19, NARS, NDUFS2, P2RX7, PDCL, MAPK13, PTGIR, PTPRA, RELA, CCL7, CCL8, CCL19, CCL22, SRC, STX4, TCEB1, TFRC, AGPS, MARCO, SNX3, CD84, USP14, ITGB1BP1, ATP6V1F, TRIP4, CD163, CIAO1, WTAP, ARHGEF11, ABI1, SCAMP2, ACTR2, BCAP31, ZMPSTE24, BCKDK, EXOC5, STIP1, UQCR11, SDS, LILRB4, OGF, TFE, FKBP15, DNAJC13, TDRD7, STX12, IL17RA, ABTB2, FAM32A, SIGLEC7, SIGLEC9, ADAMDEC1, CECR5, SLC25A24, NRBP1, MS4A4A, TREM2, OTUD4, PQLC2, HAUS2, ARL8B, NECAP2, WDR11, ZC3H15, CCDDC47, UTP3, MRS2, HAMP, MRPL40, VPS33A, CORO7, LIMD2, TMX1, DOT1L, ADO, and ADCK2, (d) ACP2, ADRA2B, ALCAM, TSPO, C3AR1, DAGLA, CALR, CHIT1, CYBB, CYC1, CYP19A1, DLAT, FCER1G, GP1BA, GPD1, IFNAR1, IL10, KCNJ5, KIFC3, MT2A, MYBPH, MYH11, MYO7A, P2RX7, PRDX1, RAB3IL1, RNH1, MRPL12, CCL1, CCL7, CCL8, CCL24, SRC, VIM, RRP1, MARCO, S1PR2, AP1M2, ACTR3, LILRB1, AFG3L2, SDS, LILRB4, EMILIN1, VSIG4, HSPB7, COQ2, ADAMDEC1, CECR5, WSB2, SLAMF8, DNASE2B, CLPB, MFSD7, and ADCK2, (SAM2、SAMA33、SAMA20、SAMA2、SAS1 FAMOUS、FAMOUS、FAMOUS、FAMOUS33 C1、SYS11、SYS1、SYS3、SYS1 1101、CHY10、CHY1、CHY3、CHY 100001000000000000000000000000000000 33、HAR10、HAR10、HYS1、DY20、DYS 20207、00000000000000000000000000000000000000000000000. ROSE7、ROSE8、ROSE18、ROSE19、ROSE24、ROSE1 、SAKA、SAX、SAS1、SASHA、SHA201、SAS22、S 4. 163、LOVE1、DY102、CHSIS、LOVES10、SHR、4 LOVE 4、 LOVE1 1 LOVE1 CHEEK1、HARSH15、HARSHY、HARHYH、HARSHAR4、HARSH 11、DYS2、DYSYS1、DYSYS1、DYSYS 10、SHAS10、SHAS2、SHAS20、SHAS70、SHAS C2、DY33、SYSY8、SYSY、DYSYSY、F1 11. LOVE3, LOVE4, and LOVE2 (f) ACP2, ADCY3, ADRA2B, ALCAM, ABCD1, ANXA2, ATP6V1A, C1QA, C1QB, C3AR1, DAGLA, CD80, CD63, CHIT1, CMKLR1, SLC31A1, CSF1, CSF1R, CYBB, CYC1, CYP19A 1, FANCE, FDX1, FPR2, FPR3, GPD1, HEXB, KCNJ1, KCNJ5, KIFC3, MMP19, MSR1, M T2A, MYBPH, P2RX7, MAPK13, S100A11, CCL1, CCL7, CCL8, CCL18, CCL19, CCL22, CCL24, SLC1A2, SLC6A12, SLC11A1, SIGLEC1, SRC, TIE1, MARCO, HYAL2, CD163 , LONP1, IGSF6, LILRB1, CD300C, SDS, LILRB4, EMILIN1, VSIG4, PHLDB1, NCAPH , CLEC4E, MYOF, HSPB7, ADAMDEC1, GLRX2, MS4A4A, ATP6V1H, TREM2, TMEM70, T MEM33, KCNK13, SLAMF8, HAMP, DNASE2B, MYOZ1, MFSD7, ADO, ADCK2, and TBC1D16 The immunoconjugate and anti-CD20 antibody for use according to claim 184 or 185.

187. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 182, wherein the macrophage biomarker is the mean of the tumor-associated macrophage gene signature set score of one or more tumor-associated macrophage gene signature sets.

188. The immunoconjugate and anti-CD20 antibody for use according to claim 187, wherein each tumor-associated macrophage gene signature set score is the average of the expression levels of one or more genes in the tumor-associated macrophage gene signature set.

189. The immunoconjugate and anti-CD20 antibody for use according to claim 188, wherein each tumor-associated macrophage gene signature set score is the mean of the normalized expression levels of one or more genes in the tumor-associated macrophage gene signature set.

190. The aforementioned set of one or more tumor-associated macrophage gene signatures (a) MARCO, ACP5, VSIG4, MRC1, MSR1, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, C1QC, APOE, FOLR2, CTSD and SPP1 The immunoconjugate and anti-CD20 antibody for use according to claim 188 or 189.

191. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 182, wherein the macrophage biomarker is a gene expression value.

192. The immunoconjugate and anti-CD20 antibody for use according to claim 187, wherein the gene expression value is the median gene expression value.

193. The immunoconjugate and anti-CD20 antibody for use according to claim 187 or 192, wherein the gene expression level is measured using a gene signature matrix.

194. The aforementioned gene signature matrix includes the following genes: (a) CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4 , PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, F ASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5 B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBTB16, FOX P3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM46C, B LK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, K LRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL 11A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, HOPX, CD1D, GNG7, TC F4, BANK1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLEKH F1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LR RC32, ARHGAP24, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, AKAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PTG S1, TMEM170B, TREM2, ECM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2 A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CF B, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, C XCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1, or (b)CD200、KLHL14、TCL1A、NRG1、CYP4 F3、EOMES、PPP2R2B、RNF165、WNT7A、C CR4、PDGFD、EBF1、FCGBP、PCDH9、MLC1 、TSHZ2、S1PR5、NCALD、LAYN、CD248、GC NT4、FASLG、TRAT1、ADAM6、GUCY1A3、L RRC4、TSPAN18、SBK1、ICOS、BTNL8、WN T5B、AUTS2、SH2D2A、ADGRG3、PNOC、SPIB、VPREB3、DPEP3、MME、ZBTB16、FOXP3 、SEMA3G、CD8A、TOGARAM2、COLGALT2、 ABCB1、STAP1、SAMD3、FAM46C、BLK、CT LA4、CD19、REPS2、RTKN2、POU2AF1、DA PK2、PYHIN1、NLRC3、GATM、KLRD1、AFF3 、FCRLA、AATBC、REM2、YPEL1、TXK、CD8 B、P2RX5、CEACAM1、BCL11A、NINJ2、AB CB4、CD5、HAL、HPGD、BLNK、PLCL1、CEP 19、HPSE、SLFN13、HOPX、CD1D、GNG7、TM EM154、TCF4、BANK1、FHIT、FCMR、GNG2 、GFRA2、KBTBD11、TECPR2、RALGPS2、T SPOAP1、PLEKHF1、MEF2C、MAOA、TTYH2 、HLA-DOB、NRGN、DGAT2、FXYD6、TMCC3、 MGAM、TTC38、LRRC32、ARHGAP24、PPP1 R3B、STAT4、SLC7A8、CD72、FZD1、GK5、 DYSF、PLTP、SMARCD3、FAM160B1、PDPN 、AKAP2、ACVRL1、KCNJ15、CD36、ALDH1A 2、ENPP2、COLEC12、PTGS1、TMEM170B、 DOCK5、TREM2、C5AR2、ECM1、SLC1A3、A BHD5、MS4A4A、CLIC2、IL1R1、SLC2A6、 GAS7、RNF144B、SLC6A12、FPR2、ADAM28 、GRK3、KDM1B、MATK、LMO2、CFB、CCRL2 、CLEC4A、TLR4、LILRA2、ACE、TLR1、LRR K2、LY96、NUPR1、CISH、CSTA、EREG、AD AMDEC1、RNASE6、CXCL3、VSIG4、CXCL2、CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1, The immunoconjugate and anti-CD20 antibody for use according to claim 193, comprising:

195. The gene signature matrix described above includes the following genes: CD200, KLHL14, TCL1A, NRG1, EOMES, PPP2R2B, RNF165, WNT7A, CCR4, PDGFD, EBF1, FCGBP, PCDH9, MLC1, TSHZ2, S1PR5, NCALD, LAYN, GCNT4, FASLG, TRAT1, ADAM6, GUCY1A3, LRRC4, TSPAN18, SBK1, ICOS, BTNL8, WNT5B, AUTS2, SH2D2A, ADGRG3, PNOC, SPIB, VPREB3, DPEP3, MME, ZBT B16, FOXP3, SEMA3G, CD8A, TOGARAM2, COLGALT2, ABCB1, STAP1, SAMD3, FAM4 6C, BLK, CTLA4, CD19, REPS2, RTKN2, POU2AF1, DAPK2, PYHIN1, NLRC3, GATM, KLRD1, AFF3, FCRLA, AATBC, REM2, YPEL1, TXK, CD8B, P2RX5, CEACAM1, BCL11 A, ABCB4, CD5, HPGD, BLNK, PLCL1, HPSE, SLFN13, HOPX, CD1D, GNG7, TCF4, BAN K1, FHIT, FCMR, GNG2, GFRA2, KBTBD11, RALGPS2, TSPOAP1, PLEKHF1, MEF2C, MAOA, TTYH2, HLA-DOB, DGAT2, FXYD6, TMCC3, MGAM, TTC38, LRRC32, ARHGAP2 4, STAT4, SLC7A8, CD72, FZD1, GK5, DYSF, PLTP, SMARCD3, FAM160B1, PDPN, A KAP2, ACVRL1, KCNJ15, ALDH1A2, ENPP2, COLEC12, PTGS1, TMEM170B, TREM2, E An immunoconjugate and anti-CD20 antibody for use according to claim 182, comprising CM1, SLC1A3, ABHD5, MS4A4A, CLIC2, IL1R1, SLC2A6, GAS7, RNF144B, SLC6A12, FPR2, ADAM28, GRK3, KDM1B, MATK, LMO2, CFB, CCRL2, CLEC4A, LILRA2, ACE, NUPR1, CISH, EREG, ADAMDEC1, RNASE6, CXCL3, VSIG4, CXCL2, CD86, LILRB4, SERPING1, SQOR, INHBA, and ICAM1.

196. An immunoconjugate and an anti-CD20 antibody for use according to any one of claims 193 to 195, wherein the gene signature matrix is ​​used to determine the number of M1 macrophages or tumor-associated macrophages.

197. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 182, wherein the macrophage biomarker is the amount of M1 macrophages or tumor-associated macrophages.

198. The immunoconjugate and anti-CD20 antibody for use according to claim 197, wherein the amount of M1 macrophages or tumor-associated macrophages is measured directly or indirectly.

199. The immunoconjugate and anti-CD20 antibody for use according to claim 198, wherein the amount of M1 macrophages or tumor-associated macrophages is directly measured using flow cytometry, spatial transcriptomics, spatial proteomics, or a combination thereof.

200. The immunoconjugate and anti-CD20 antibody for use according to claim 198, wherein the amount of M1 macrophages or tumor-associated macrophages is measured indirectly using nucleic acids or proteins.

201. The immunoconjugate and anti-CD20 antibody for use according to claim 200, wherein the nucleic acid is measured using RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

202. The immunoconjugate and anti-CD20 antibody for use according to claim 201, wherein the amount of M1 macrophages or tumor-associated macrophages is measured using a marker gene approach or a deconvolution approach.

203. The immunoconjugate and anti-CD20 antibody for use according to claim 202, wherein the marker gene approach uses xCell.

204. The deconvolution approach uses quanTIseq, and the immunoconjugate and anti-CD20 antibody for use according to claim 202.

205. An immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 182, wherein macrophage biomarkers in the sample from the patient are measured using nucleic acids or proteins.

206. The immunoconjugate and anti-CD20 antibody for use according to claim 205, wherein the macrophage biomarker in the sample from the patient is determined using nucleic acid expression levels.

207. The immunoconjugate and anti-CD20 antibody for use according to claim 206, wherein the nucleic acid expression level is determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

208. The immunoconjugate and anti-CD20 antibody for use according to claim 206 or 207, wherein the nucleic acid expression level is the mRNA expression level.

209. The immunoconjugate and anti-CD20 antibody for use according to claim 208, wherein the mRNA expression level is determined by RNA-seq.

210. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 209, wherein the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

211. The immunoconjugate and anti-CD20 antibody for use according to claim 210, wherein the sample is a tissue sample.

212. The immunoconjugate and anti-CD20 antibody for use according to claim 211, wherein the tissue sample is a tumor tissue sample.

213. The immunoconjugate and anti-CD20 antibody for use according to claim 212, wherein the tumor tissue sample comprises tumor cells, tumor-infiltrating immune cells, stromal cells, normal adjacent tissue (NAT) cells, or a combination thereof.

214. The immunoconjugate and anti-CD20 antibody for use according to claim 212 or 213, wherein the tumor tissue sample is a biopsy material.

215. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 210 to 214, wherein the sample is an archived sample, a fresh sample, or a frozen sample.

216. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 215, wherein the DLBCL is a germinal center B cell-like (GCB) or activated B cell-like (ABC) originating cell subgroup of DLBCL.

217. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 216, wherein the DLBCL is CD79a-positive DLBCL and / or CD20-positive DLBCL.

218. An immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 217, wherein the patient has not been previously treated for DLBCL.

219. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 218, wherein the patient has not been previously administered the immunoconjugate and the anti-CD20 antibody.

220. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 219, wherein the anti-CD79b antibody comprises a heavy chain variable domain (VH) containing the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) containing the amino acid sequence of SEQ ID NO:

4.

221. The aforementioned anti-CD79b antibody, (a) A heavy chain containing the amino acid sequence of SEQ ID NO: 13 and a light chain containing the amino acid sequence of SEQ ID NO: 11, or (b) A heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 14, or (c) Heavy chain containing the amino acid sequence of SEQ ID NO: 12 and light chain containing the amino acid sequence of SEQ ID NO: 11 An immunoconjugate and an anti-CD20 antibody for use according to any one of claims 168 to 220, comprising:

222. An immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 221, wherein p is 2 to 7, 2 to 6, 2 to 5, 3 to 5, or 3 to 4.

223. An immunoconjugate and anti-CD20 antibody for use according to claim 222, wherein p is 3.

4.

224. An immunoconjugate and anti-CD20 antibody for use according to claim 222, wherein p is 3.

5.

225. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 224, wherein the immunoconjugate is polatuzumab vedotin.

226. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 225, wherein the anti-CD20 antibody is a type I anti-CD20 antibody or a type II anti-CD20 antibody.

227. The immunoconjugate and anti-CD20 antibody for use according to claim 226, wherein the anti-CD20 antibody is a type I anti-CD20 antibody.

228. The aforementioned type I anti-CD20 antibody is the following CDR: (a) CDR-H1 having the amino acid sequence of SEQ ID NO: 26, (b) CDR-H2 having the amino acid sequence of SEQ ID NO: 27, (c) CDR-H3 having the amino acid sequence of SEQ ID NO: 28, (d) CDR-L1 having the amino acid sequence of SEQ ID NO: 29, (e) CDR-L2 having the amino acid sequence of SEQ ID NO: 30, (f) CDR-L3 having the amino acid sequence of SEQ ID NO: 31 The immunoconjugate and anti-CD20 antibody for use according to claim 227, comprising:

229. The immunoconjugate and anti-CD20 antibody for use according to claim 228, wherein the type I anti-CD20 antibody comprises a VH domain containing the amino acid sequence of SEQ ID NO: 40 and a VL domain containing the amino acid sequence of SEQ ID NO:

41.

230. The immunoconjugate and anti-CD20 antibody for use according to claim 229, wherein the type I anti-CD20 antibody is rituximab.

231. An immunoconjugate and anti-CD20 antibody for use according to any one of claims 225 to 230, wherein polatuzumab vedotin is for use in a dose of about 1.0 mg / kg to about 1.8 mg / kg.

232. The immunoconjugate and anti-CD20 antibody for use according to claim 231, wherein polatuzumab vedotin is for use at a dose of approximately 1.8 mg / kg.

233. Rituximab at approximately 375 mg / m² ２ An immunoconjugate and an anti-CD20 antibody for use according to any one of claims 230 to 232, for use in the specified dose.

234. An immunoconjugate and anti-CD20 antibody for use according to any one of claims 225 to 233, wherein polatuzumab vedotin and / or rituximab are for intravenous use.

235. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 168 to 234, wherein the treatment further comprises the use of an effective amount of an additional therapeutic agent.

236. The immunoconjugate and anti-CD20 antibody for use according to claim 235, wherein the additional therapeutic agent is one or more of the following: chemotherapeutic agents, corticosteroids, antineoplastic agents, growth inhibitors, anti-angiogenic agents, radiotherapy, cytotoxic agents, or combinations thereof.

237. The immunoconjugate and anti-CD20 antibody for use according to claim 236, wherein the additional therapeutic agent is a chemotherapeutic agent and a corticosteroid.

238. The immunoconjugate and anti-CD20 antibody for use according to claim 236 or 237, wherein the chemotherapeutic agent is cyclophosphamide and / or doxorubicin.

239. The immunoconjugate and anti-CD20 antibody for use according to any one of claims 236 to 238, wherein the corticosteroid is prednisone, prednisolone, or methylprednisolone.

240. Cyclophosphamide is approximately 375 mg / m². ２ ~Approx. 750mg / m ２ An immunoconjugate and an anti-CD20 antibody for use according to claim 238 or 239, for use in the specified dose.

241. Doxorubicin at approximately 25 mg / m² ２ ~Approx. 50mg / m ２ An immunoconjugate and an anti-CD20 antibody for use according to any one of claims 238 to 240, for use in the specified dose.

242. (a) Prednisone is intended for use in doses of approximately 100 mg, or (b) Prednisolone is intended for use in doses of approximately 100 mg, or (c) This is intended for use at a dose of approximately 80 mg of methylprednisolone. An immunoconjugate and an anti-CD20 antibody for use according to any one of claims 239 to 241.

243. An immunoconjugate and anti-CD20 antibody for use according to any one of claims 238 to 242, wherein cyclophosphamide and / or doxorubicin are for intravenous use.

244. An immunoconjugate and an anti-CD20 antibody for use according to any one of claims 239 to 243, wherein prednisone, prednisolone, or methylprednisolone is for oral use.

245. An immunoconjugate and anti-CD20 antibody for use according to any one of claims 225 to 244, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin and / or prednisone, prednisolone, or methylprednisolone are for use in at least one 21-day cycle.

246. (a) Polatuzumab vedotin, rituximab, cyclophosphamide and / or doxorubicin are for use on day 1 of each 21-day cycle, and / or (b) Prednisone, prednisolone, or methylprednisolone for use on days 1 through 5 of each 21-day cycle. An immunoconjugate and an anti-CD20 antibody for use according to claim 245.

247. Immunoconjugates and anti-CD20 antibodies for use according to claim 245 or 246, wherein polatuzumab vedotin, rituximab, cyclophosphamide, doxorubicin, and / or prednisone, prednisolone, or methylprednisolone are for use in one, two, three, four, five, or six 21-day cycles.

248. A method for identifying, diagnosing and / or predicting whether a patient with diffuse large B-cell lymphoma (DLBCL) can benefit from a treatment comprising an immunoconjugate, an anti-CD20 antibody, a chemotherapeutic agent and a corticosteroid, comprising measuring a macrophage biomarker in a sample from the patient, wherein the amount or level of the macrophage biomarker in the sample is below a reference macrophage biomarker amount or level, thereby identifying, diagnosing and / or predicting the patient to benefit from the treatment comprising the immunoconjugate, the anti-CD20 antibody, the chemotherapeutic agent and the corticosteroid, wherein the immunoconjugate is of the following formula: 【Transformation 8】 (In the formula, Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5) The aforementioned anti-CD20 antibody is rituximab. The aforementioned chemotherapeutic agent comprises cyclophosphamide and doxorubicin. The corticosteroid includes prednisone. method.

249. A method for selecting a therapy for a patient having DLBCL, comprising measuring a macrophage biomarker in a sample from the patient, wherein the amount or level of the macrophage biomarker in the sample is below a reference amount or level of macrophage biomarker, thereby identifying the patient as being able to benefit from a treatment comprising an immunoconjugate, an anti-CD20 antibody, a chemotherapeutic agent and a corticosteroid, wherein the immunoconjugate is of the following formula: 【Chemistry 9】 (In the formula, Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5) The aforementioned anti-CD20 antibody is rituximab. The aforementioned chemotherapeutic agent comprises cyclophosphamide and doxorubicin. The corticosteroid includes prednisone. method.

250. The method according to claim 248 or 249, wherein the amount or level of the macrophage biomarker from the patient is below the reference amount or level of the macrophage biomarker, and the method further comprises administering to the patient an effective amount of the immunoconjugate, an effective amount of the anti-CD20 antibody, an effective amount of the chemotherapeutic agent, and an effective amount of the corticosteroid.

251. A method for treating a patient with DLBCL, (a) Measuring macrophage biomarkers in a sample from the patient, wherein the amount or level of macrophage biomarkers in the sample is below the reference amount or level of macrophage biomarkers, and (b) Based on the macrophage biomarker measured in step (a), administer to the patient an effective amount of immunoconjugate, an effective amount of anti-CD20 antibody, an effective amount of chemotherapeutic agent and an effective amount of corticosteroid, wherein the immunoconjugate has the following formula: 【Chemistry 10】 (In the formula, Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5) The aforementioned anti-CD20 antibody is rituximab. The aforementioned chemotherapeutic agent comprises cyclophosphamide and doxorubicin. The corticosteroid includes prednisone. method.

252. A method for treating a patient with DLBCL, comprising administering to the patient an effective amount of an immunoconjugate, an effective amount of an anti-CD20 antibody, an effective amount of a chemotherapeutic agent, and an effective amount of a corticosteroid, wherein, prior to treatment, it has been determined that the amount or level of a macrophage biomarker in a sample from the patient is below a reference amount or level of a macrophage biomarker, and the immunoconjugate has the following formula: 【Chemistry 11】 (In the formula, Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5) The aforementioned anti-CD20 antibody is rituximab. The aforementioned chemotherapeutic agent comprises cyclophosphamide and doxorubicin. The corticosteroid includes prednisone. method.

253. A method for treating a patient having DLBCL and in which the amount or level of a macrophage biomarker in a patient sample is below the reference amount or level of a macrophage biomarker, comprising administering to the patient an effective amount of an immunoconjugate, an effective amount of an anti-CD20 antibody, an effective amount of a chemotherapeutic agent and an effective amount of a corticosteroid, wherein the immunoconjugate has the following formula: 【Chemistry 12】 (In the formula, Ab is an anti-CD79b antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 12 and a light chain containing the amino acid sequence of SEQ ID NO: 11, and p is 3.5) The aforementioned anti-CD20 antibody is rituximab. The aforementioned chemotherapeutic agent comprises cyclophosphamide and doxorubicin. The corticosteroid includes prednisone. method.

254. The method according to any one of claims 248 to 253, wherein the immunoconjugate is polatuzumab vedotin.

255. The immunoconjugate is administered at a dose of approximately 1.8 mg / kg, and the rituximab is administered at a dose of approximately 375 mg / m². ２ The dose is administered, and the cyclophosphamide is approximately 750 mg / m². ２ The dose administered is approximately 50 mg / m² of doxorubicin. ２ The method according to any one of claims 248 to 254, wherein the drug is administered in a dose of approximately 100 mg of the prednisone.

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