Method for determining therapeutic effect of lung cancer treatment comprising anti-PD-L1 antagonist and anti-TIGHT antagonist antibodies

Abstract

The present invention relates to prognosis and therapeutic methods for the treatment of cancer (e.g., lung cancer, e.g., non-small cell lung cancer (NSCLC)) using the expression levels of tumor-associated macrophage (TAM) and regulatory T cell (Treg) genes. In particular, the present invention provides methods for patient selection and treatment.

Description

[0001] sequence list

[0002] This application contains a sequence list, which has been submitted electronically in XML format and is incorporated herein by reference in its entirety. The XML copy was created on June 1, 2023, named 50474-290WO4_Sequence_Listing_6_1_23, and is 33,392 bytes in size. Technical Field

[0003] This article provides prognostic and therapeutic approaches for treating cancers (e.g., lung cancer, such as non-small cell lung cancer (NSCLC)) using the expression levels of tumor-associated macrophages (TAMs) and regulatory T-cell (Treg) genes. In particular, the invention provides methods for patient selection and treatment. Background Technology

[0004] Cancer is characterized by the uncontrolled growth of subpopulations of cells. It is the leading cause of death in developed countries and the second leading cause of death in developing countries, with over 14 million new cancer diagnoses and over 8 million cancer deaths annually. Therefore, cancer care represents a significant and ever-increasing social burden.

[0005] Programmed cell death-1 / programmed cell death ligand-1 (PD-1 / PD-L1) blockade is effective against a variety of malignancies. However, not all patients benefit, and a large proportion of those who initially respond eventually relapse. One approach to prolonging and expanding the effects of cancer immunotherapy has been to target additional immune checkpoints. One such co-inhibitory checkpoint is TIGIT (a T-cell immune receptor with Ig and immune receptor tyrosine inhibitory motif (ITIM) domains).

[0006] Non-small cell lung cancer (NSCLC) is the major subtype of lung cancer, accounting for approximately 80%-85% of all cases. For advanced disease, the overall five-year survival rate is 2%-4%.

[0007] Although first-line treatment for patients with advanced NSCLC has improved, leading to longer survival and reduced disease-related symptoms, almost all patients experience disease progression. Cancer immunotherapy, in particular, offers the possibility of long-term disease control. Specifically, NSCLC patients have been found to benefit from treatment using a combination of a PD-1 axis-binding antagonist (atezilzumab) and an anti-TIGIT antagonist antibody (tisrelimumab).

[0008] Therefore, there is an unmet need in the field for a robust prognostic approach that identifies patients who may benefit from treatments including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies to more effectively manage their disease. Summary of the Invention

[0009] In one aspect, the present invention provides a method for identifying an individual with cancer who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, the method comprising detecting the expression levels of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from the individual and determining a tumor-associated macrophage (TAM) eigenvalue score therefrom, wherein a TAM eigenvalue score higher than a reference TAM eigenvalue score identifies the individual as one who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0010] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer, the method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a TAM characteristic score therefrom, wherein a TAM characteristic score higher than a reference TAM characteristic score identifies the individual as an individual who may benefit from treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

[0011] In some respects, the individual has a TAM characteristic score higher than the reference TAM characteristic score in the sample, and the method further includes administering an effective amount of PD-1 axis-binding antagonist and anti-TIGIT antagonist antibody to the individual.

[0012] On the other hand, the present invention provides a method for treating an individual suffering from cancer, the method comprising (a) detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a TAM characteristic score therefrom, wherein the TAM characteristic score is higher than a reference TAM characteristic score, thereby identifying the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody; and (b) administering an effective amount of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody to the individual.

[0013] On the other hand, the present invention provides a method for treating an individual suffering from cancer, the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, wherein the individual has been determined to have a TAM characteristic score higher than a reference TAM characteristic score, thereby identifying the individual as an individual who may benefit from treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, and wherein the TAM characteristic score is based on the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO detected in a sample from the individual.

[0014] In some respects, samples were obtained from individuals prior to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies.

[0015] In some respects, the benefit is an increase in progression-free survival (PFS), objective response rate (ORR), or overall survival (OS).

[0016] In some respects, the reference TAM feature scores are pre-specified TAM feature scores.

[0017] In some respects, the reference TAM characteristic score is the TAM characteristic score in the reference population. In some respects, the TAM characteristic score in the reference population is the median TAM characteristic score of the reference population. In some respects, the reference population is a group of individuals with cancer.

[0018] In some respects, the TAM characteristic score is the average expression level of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in samples from individuals.

[0019] In some respects, the method includes further detecting the expression levels of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in samples from individuals.

[0020] In some respects, the TAM characteristic score is the average of the expression levels of one or more of the following individuals in a sample: C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, and ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FORL2, and CTSD.

[0021] In some aspects, the method includes further detecting the expression levels of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in samples from an individual, and determining a TAM characteristic score from it, wherein the TAM characteristic score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in samples from an individual.

[0022] In some respects, the expression levels of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD have been detected in samples from individuals.

[0023] In some respects, the TAM characteristic score is the average expression level of one or more of the following in samples from an individual: C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, and ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD.

[0024] In some respects, the expression levels of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD have been detected in samples from individuals and TAM characteristic scores have been determined from them, where the TAM characteristic score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in samples from individuals.

[0025] On the other hand, the present invention provides a method for monitoring the response of an individual with cancer to treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, the method comprising detecting, at time points during or after administration of the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody, MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, and LYA in samples from the individual. The expression levels of one or more of M1, LCAT, and LIRA3, wherein an increase in the expression level of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 relative to their respective reference expression levels predicts individuals who may respond to the treatment, including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies.

[0026] In some respects, the expression levels of one or more of the following were measured three weeks after the start of treatment, which included PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies: MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3.

[0027] In some respects, the expression levels of one or more of the following were measured six weeks after the initiation of treatment, which included PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies: MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3.

[0028] In some respects, the expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 are increased relative to their respective reference expression levels, thereby predicting that an individual may respond to treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies, and the method further includes administering additional doses of PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies to the individual.

[0029] In some respects, the response to treatment is an increase in PFS or OS.

[0030] In some respects, the reference expression level is the baseline expression level in an individual's sample at a time point prior to the initiation of treatment, which includes both PD-1 axis-binding antagonists and anti-TIGIT antagonist antibodies.

[0031] On the other hand, the present invention provides a method for identifying individuals with cancer who may benefit from treatment including PD-1 axis-binding antagonists and anti-TIGIT antagonist antibodies, the method comprising detecting the expression levels of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual and determining a regulatory T cell (Treg) eigenvalue score therefrom, wherein a Treg eigenvalue score higher than a reference Treg eigenvalue score identifies the individual as one who may benefit from treatment including PD-1 axis-binding antagonists and anti-TIGIT antagonist antibodies.

[0032] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer, the method comprising detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a Treg eigenvalue score therefrom, wherein a Treg eigenvalue score higher than a reference Treg eigenvalue score identifies the individual as an individual who may benefit from treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

[0033] In some respects, the individual has a Treg characteristic score higher than that of a reference Treg characteristic score in the sample, and the method further includes administering an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody to the individual.

[0034] On the other hand, the present invention provides a method for treating an individual suffering from cancer, the method comprising (a) detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a Treg characteristic score therefrom, wherein the Treg characteristic score is higher than a reference Treg characteristic score, thereby identifying the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody; and (b) administering an effective amount of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody to the individual.

[0035] On the other hand, the present invention provides a method for treating an individual with cancer, the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, wherein the individual has been determined to have a Treg eigenvalue score higher than a reference Treg eigenvalue score, thereby identifying the individual as an individual who may benefit from treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, and wherein the Treg eigenvalue score is based on the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in a sample from the individual.

[0036] In some respects, samples were obtained from individuals prior to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies.

[0037] In some respects, the benefits come from improvements in PFS, ORR, or OS.

[0038] In some respects, the reference Treg feature scores are pre-specified Treg feature scores.

[0039] In some respects, the reference Treg characteristic score is the Treg characteristic score in the reference population. In some respects, the Treg characteristic score in the reference population is the median Treg characteristic score of the reference population. In some respects, the reference population is a group of individuals with cancer.

[0040] In some respects, the Treg characteristic score is the mean of the expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in samples from individuals.

[0041] In some respects, the expression level is either the nucleic acid expression level or the protein expression level.

[0042] In some respects, expression levels are defined as nucleic acid expression levels. In others, nucleic acid expression levels are determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or combinations thereof.

[0043] In some respects, nucleic acid expression levels are equivalent to mRNA expression levels. In other respects, mRNA expression levels are determined using RNA-seq.

[0044] In some respects, expression level is the same as protein expression level. In other respects, protein expression level is determined by mass spectrometry.

[0045] In some respects, the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

[0046] In some respects, the sample is a tissue sample. In some respects, the tissue sample is a tumor tissue sample. In some respects, the tumor tissue sample is a biopsy section.

[0047] In some respects, the sample is a serum sample.

[0048] In some respects, the samples are archived samples, fresh samples, or frozen samples.

[0049] In some respects, the samples have been identified by immunohistochemistry (IHC) as having a fraction of PD-L1 positive tumor cells.

[0050] In some respects, the fraction of PD-L1 positive tumor cells is determined by positive staining with anti-PD-L1 antibodies, such as SP263, 22C3, SP142, or 28-8.

[0051] In some respects, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, determined by positive staining with anti-PD-L1 antibody SP263. In other respects, the PD-L1 positive tumor cell fraction is calculated using Ventana SP263 IHC assay.

[0052] In some respects, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, determined by positive staining with anti-PD-L1 antibody 22C3. In other respects, the PD-L1 positive tumor cell fraction is calculated using pharmDx 22C3 IHC assay.

[0053] In some respects, the cancer is lung cancer. In other respects, lung cancer is non-small cell lung cancer (NSCLC).

[0054] In some respects, the anti-TIGIT antagonist antibody contains the following hypervariable regions (HVR): (a) HVR-H1, which contains the amino acid sequence SNSAAWN (SEQ ID NO:1); (b) HVR-H2, which contains the amino acid sequence KTYYRFKWYSDYAVSVKG (SEQ ID NO:2); (c) HVR-H3, which contains the amino acid sequence ESTTYDLLAGPFDY (SEQ ID NO:3); (d) HVR-L1, which contains the amino acid sequence KSSQTVLYSSNNKKYLA (SEQ ID NO:4); (e) HVR-L2, which contains the amino acid sequence WASTRES (SEQ ID NO:5); and (f) HVR-L3, which contains the amino acid sequence QQYYSTPFT (SEQ ID NO:6).

[0055] In some respects, the anti-TIGIT antagonist antibody further comprises the following light chain variable regions FR: (a) FR-L1 comprising the amino acid sequence DIVMTQSPDSLAVSLGERATINC (SEQ ID NO:7), (b) FR-L2 comprising the amino acid sequence WYQQKPGQPPNLLIY (SEQ ID NO:8), and (c) FR-L3 comprising the amino acid sequence GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO:9); and (d) FR-L4 comprising the amino acid sequence FGPGTKVEIK (SEQ ID NO:10).

[0056] In some respects, the anti-TIGIT antagonist antibody further comprises the following heavy chain variable region FR: (a) FR-H1 comprising the amino acid sequence X1VQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO:11), wherein X1 is Q or E; (b) FR-H2 comprising the amino acid sequence WIRQSPSRGLEWLG (SEQ ID NO:12); (c) comprising

[0057] FR-H3 containing the amino acid sequence RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO:13); and (d) FR-H4 containing the amino acid sequence WGQGTLVTVSS (SEQ ID NO:14).

[0058] In some respects, X1 is Q. In other respects, X1 is E.

[0059] In some respects, the anti-TIGIT antagonist antibody comprises (a) a VH domain containing an amino acid sequence having at least 95% sequence identity with the amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWL GKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCT RESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO:17) or QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWL GKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO:18);

[0060] (b) The VL structural domain, which contains and

[0061] DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAWYQQKPGQP

[0062] The amino acid sequence of PNLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYS TPFTFGPGTKVEIK (SEQ ID NO:19) has an amino acid sequence with at least 95% sequence identity; or (c) the VH domain as in (a) and the VL domain as in (b).

[0063] In some respects, the anti-TIGIT antagonist antibody comprises (a) a VH domain containing the amino acid sequence of SEQ ID NO:17 or 18; and (b) a VL domain containing the amino acid sequence of SEQ ID NO:19.

[0064] In some respects, the anti-TIGIT antagonist antibody comprises (a) a VH domain containing the amino acid sequence of SEQ ID NO:17; and (b) a VL domain containing the amino acid sequence of SEQ ID NO:19.

[0065] In some respects, the anti-TIGIT antagonist antibody comprises: (a) a heavy chain containing the amino acid sequence of SEQ ID NO:33, and (b) a light chain containing the amino acid sequence of SEQ ID NO:34.

[0066] In some respects, anti-TIGIT antagonist antibodies are monoclonal antibodies.

[0067] In some respects, anti-TIGIT antagonist antibodies are human antibodies.

[0068] In some respects, anti-TIGIT antagonist antibodies are full-length antibodies.

[0069] In some respects, anti-TIGIT antagonist antibodies exhibit effector functions.

[0070] In some respects, anti-TIGIT antagonist antibodies contain an Fc domain that can interact with the Fcγ receptor (FcγR).

[0071] In some respects, anti-TIGIT antagonist antibodies are IgG class antibodies. In other respects, IgG class antibodies are IgG1 subclass antibodies.

[0072] In some respects, the anti-TIGIT antagonist antibody is tireliumab.

[0073] In some respects, anti-TIGIT antagonist antibodies are antibody fragments selected from the group consisting of Fab, Fab', Fab'-SH, Fv, single-chain variable fragments (scFv), and (Fab')2 fragments that bind to TIGIT.

[0074] In some respects, anti-TIGIT antagonist antibodies are vimbrolizumab, atelimab, EOS084448, SGN-TGT, TJ-T6, BGB-A1217, or AB308.

[0075] In some respects, PD-1 axis binding antagonists are selected from the group consisting of PD-L1 binding antagonists, PD-1 binding antagonists and PD-L2 binding antagonists.

[0076] In some respects, PD-1 axis binding antagonists are PD-L1 binding antagonists.

[0077] In some respects, PD-L1 binding antagonists inhibit the binding of PD-L1 to one or more of its ligand-binding partners. In other respects, PD-L1 binding antagonists inhibit the binding of PD-L1 to PD-1, B7-1, or both PD-1 and B7-1.

[0078] In some respects, PD-L1 binding antagonists are anti-PD-L1 antagonist antibodies. In some respects, anti-PD-L1 antagonist antibodies include atezolizumab, MDX-1105, durvalumab, acimetab, SHR-1316, CS1001, envorimab, TQB2450, ZKAB001, LP-002, CX-072, IMC-001, KL-A167, APL-502, cochilimumab, lodalimumab, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, RC98, PDL-GEX, KD036, KY1003, YBL-007, or HS-636.

[0079] In some respects, the anti-PD-L1 antagonist antibody is atezolizumab.

[0080] In some respects, the anti-PD-L1 antagonist antibody comprises the following HVRs: (a) an HVR-H1 sequence comprising the amino acid sequence of GFTFSDSWIH (SEQ ID NO:20); (b) an HVR-H2 sequence comprising the amino acid sequence of AWISPYGGSTYYADSVKG (SEQ ID NO:21); (c) an HVR-H3 sequence comprising the amino acid sequence of RHWPGGFDY (SEQ ID NO:22); (d) an HVR-L1 sequence comprising the amino acid sequence of RASQDVSTAVA (SEQ ID NO:23); (e) an HVR-L2 sequence comprising the amino acid sequence of SASFLYS (SEQ ID NO:24); and (f) an HVR-L3 sequence comprising the amino acid sequence of QQYLYHPAT (SEQ ID NO:25).

[0081] In some respects, the anti-PD-L1 antagonist antibody comprises: (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO:26; (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 95% sequence identity with the amino acid sequence of SEQ ID NO:27; or (c) the VH domain as in (a) and the VL domain as in (b).

[0082] In some respects, the anti-PD-L1 antagonist antibody comprises: (a) a VH domain containing the amino acid sequence of SEQ ID NO:26; and (b) a VL domain containing the amino acid sequence of SEQ ID NO:27.

[0083] In some respects, the anti-PD-L1 antagonist antibody comprises: (a) a heavy chain containing the amino acid sequence of SEQ ID NO:28, and (b) a light chain containing the amino acid sequence of SEQ ID NO:29.

[0084] In some respects, anti-PD-L1 antagonist antibodies are monoclonal antibodies.

[0085] In some respects, anti-PD-L1 antagonist antibodies are humanized antibodies.

[0086] In some respects, anti-PD-L1 antagonist antibodies are full-length antibodies.

[0087] In some respects, anti-PD-L1 antagonist antibodies are antibody fragments selected from the group consisting of Fab, Fab', Fab'-SH, Fv, scFv and (Fab')2 fragments that bind to PD-L1.

[0088] In some respects, anti-PD-L1 antagonist antibodies are IgG class antibodies. In other respects, IgG class antibodies are IgG1 subclass antibodies.

[0089] In some respects, PD-1 axis binding antagonists are PD-1 binding antagonists. In some respects, PD-1 binding antagonists inhibit the binding of PD-1 to one or more of its ligand binding partners. In some respects, PD-1 binding antagonists inhibit the binding of PD-1 to PD-L1, PD-L2, or both PD-L1 and PD-L2.

[0090] In some respects, PD-1 binding antagonists are anti-PD-1 antagonist antibodies. In some respects, anti-PD-1 antagonist antibodies include nivolumab, pembrolizumab, MEDI-0680, spartazolizumab, cimiprimab, BGB-108, palolizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dotalimab, rivanlimab, sasafenib, penaplimab, CS1003, HLX10, SCT-I10A, cepalimumab, batitilimab, genolimab, BI754091, cilimab, YBL-006, BAT1306, HX008, bugalilimab, AMG 404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, AM0001, and ENUM. 244C8, ENUM 388D4, STI-1110, AK-103 or hAb21.

[0091] In some respects, PD-1 binding antagonists are Fc fusion proteins. In other respects, Fc fusion proteins are AMP-224.

[0092] In some respects, this individual is a person.

[0093] In some respects, anti-TIGIT antagonist antibodies can Fc-dependently activate bone marrow cells, optionally of which the bone marrow cells are selected from the group consisting of tumor-associated conventional dendritic cells (cDC1), macrophages, neutrophils and circulating monocytes.

[0094] In some respects, anti-TIGIT antagonist antibodies can interact with Fcγ receptors (FcγR) on bone marrow cells and can induce the mobilization of CD8+ T cells in the blood and / or the expansion of proliferating CD8+ T cells in the tumor bed.

[0095] On the other hand, the present invention provides a method for treating an individual with cancer, the method comprising administering an anti-TIGIT antagonist antibody to the individual, wherein the anti-TIGIT antagonist antibody is capable of Fc-dependent activation of bone marrow cells, optionally wherein the bone marrow cells are cells selected from the group consisting of tumor-associated conventional dendritic cells (cDC1), macrophages, neutrophils and circulating monocytes.

[0096] On the other hand, the present invention provides the use of anti-TIGIT antagonist antibodies in the manufacture of medicaments for treating cancer, wherein the anti-TIGIT antagonist antibodies are capable of Fc-dependent activation of bone marrow cells, optionally wherein the bone marrow cells are selected from the group consisting of tumor cDC1, macrophages, neutrophils and circulating monocytes.

[0097] On the other hand, the present invention provides a method for treating an individual with cancer, the method comprising administering an anti-TIGIT antagonist antibody to the individual, wherein the anti-TIGIT antagonist antibody is capable of interacting with Fcγ receptors (FcγR) on bone marrow cells and is capable of inducing mobilization of CD8+ T cells in the blood and / or expansion of proliferating CD8+ T cells in the tumor bed.

[0098] On the other hand, the present invention provides the use of anti-TIGIT antagonist antibodies in the manufacture of medicaments for treating cancer, wherein the anti-TIGIT antagonist antibodies are capable of interacting with FcγR and of inducing the mobilization of CD8+ T cells in the blood and / or the expansion of proliferating CD8+ T cells in the tumor bed.

[0099] On the other hand, the present invention provides a method for identifying individuals with cancer who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function. The method includes detecting the expression levels of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from the individual and determining a tumor-associated macrophage (TAM) characteristic score therefrom, wherein a TAM characteristic score higher than a reference TAM characteristic score identifies the individual as one who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0100] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer, the method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a TAM characteristic score therefrom, wherein a TAM characteristic score higher than a reference TAM characteristic score identifies the individual as an individual who may benefit from treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0101] In some respects, the individual has a higher TAM characteristic score than the reference TAM characteristic score in the sample, and the method further includes administering an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function to the individual.

[0102] On the other hand, the present invention provides a method for treating an individual suffering from cancer, the method comprising (a) detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from the individual and determining a TAM characteristic score therefrom, wherein the TAM characteristic score is higher than a reference TAM characteristic score, thereby identifying the individual as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function; and (b) administering to the individual an effective amount of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody exhibiting effector function.

[0103] On the other hand, the present invention provides a method for treating an individual with cancer, the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, wherein the individual has been determined to have a TAM characteristic score higher than a reference TAM characteristic score, thereby identifying the individual as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, and wherein the TAM characteristic score is based on the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO detected in a sample from the individual.

[0104] On the other hand, the present invention provides a method for monitoring the response of an individual with cancer to treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function. The method includes detecting MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, and LY in samples from the individual at time points during or after administration of the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody exhibiting effector function. The expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 relative to their respective reference expression levels predict individuals who may respond to treatment, including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies exhibiting effector functions.

[0105] On the other hand, the present invention provides a method for identifying individuals with cancer who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function. The method includes detecting the expression levels of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual and determining a regulatory T cell (Treg) eigenvalue score therefrom, wherein a Treg eigenvalue score higher than a reference Treg eigenvalue score identifies the individual as one who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0106] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer, the method comprising detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual and determining a Treg eigenvalue score therefrom, wherein a Treg eigenvalue score higher than a reference Treg eigenvalue score identifies the individual as an individual who may benefit from treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0107] In some respects, the individual has a Treg characteristic score higher than that of a reference Treg characteristic score in the sample, and the method further includes administering an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function to the individual.

[0108] On the other hand, the present invention provides a method for treating an individual suffering from cancer, the method comprising (a) detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a Treg characteristic score therefrom, wherein the Treg characteristic score is higher than a reference Treg characteristic score, thereby identifying the individual as an individual who may benefit from treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function; and (b) administering to the individual an effective amount of the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody exhibiting effector function.

[0109] On the other hand, the present invention provides a method for treating an individual with cancer, the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, wherein the individual has been determined to have a Treg characteristic score higher than a reference Treg characteristic score, thereby identifying the individual as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, and wherein the Treg characteristic score is based on the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in samples from the individual.

[0110] In some respects, anti-TIGIT antagonist antibodies contain an Fc domain that can interact with the Fcγ receptor (FcγR). BRIEF DESCRIPTION OF THE DRAWINGS

[0111] Figure 1AThis is a set of Kaplan-Meier (KM) curves showing overall survival (OS) in non-small cell lung cancer (NSCLC) patients treated with atezolizumab and placebo or tira and atezolizumab in the biomarker-evaluable population (BEP) of the CITYSCAPE trial (GO40290). Hazard ratios (HR) and 95% confidence intervals were determined using a univariate Cox model. Mo: Month.

[0112] Figure 1B This is a forest plot showing the association between high abundance of the indicated cell type in the tumor and the objective response rate (ORR) in the BEP of the CITYSCAPE trial. T+A: tisreliumab + atezolizumab. P+A: placebo + atezolizumab. Intratumoral cell types were determined as high or low based on the median eigenvalue cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0113] Figure 1C This is a set of micrographs showing H&E staining and multiplex immunofluorescence (mIF) staining of panCK (green), FoxP3 (white), CD68 (red), and programmed death-ligand 1 (PD-L1) (yellow) in tumor samples from CITYSCAPE patients (representing Treg-high, bone marrow-high (top); Treg-high, bone marrow-low (middle); and Treg-low, bone marrow-low (bottom) samples).

[0114] Figure 1D This is a set of KM curves showing overall survival (OS) in patients with tumors enriched (solid lines) or not enriched (dashed lines) of tumor-associated macrophages (TAMs) treated with placebo plus atezolizumab or tisrelinumab plus atezolizumab. Enrichment was determined by the median cell type characteristic score cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0115] Figure 1E This is a set of KM curves showing overall survival (OS) in patients with tumors enriched (solid lines) or not enriched (dashed lines) of regulatory T cells (Tregs) treated with placebo plus atezolizumab or tirelinumab plus atezolizumab. Enrichment was determined by the median cell type characteristic score cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0116] Figure 1F This is a set of KM curves showing overall survival (OS) in patients with tumors enriched (solid lines) or not enriched (dashed lines) with CD16 mononuclear cells, treated with placebo plus atezolizumab or tisrelinumab plus atezolizumab. Enrichment was determined by the median cell type characteristic score cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0117] Figure 1G This is a set of KM curves showing overall survival (OS) in patients with tumors enriched (solid lines) or not enriched (dashed lines) of CD8+ T effector cells (tGE8) in placebo plus atezolizumab or tirelimumab plus atezolizumab. Enrichment was determined by the median cell type characteristic score cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0118] Figure 2A These are a pair of charts showing the levels of the protein / peptide markers in serum relative to baseline on day 1 of cycle 2 (C2D1) in either the placebo plus atezolizumab group (left chart) or the atezolizumab plus tireliumab combination arm (right chart).

[0119] Figure 2B This is a heatmap showing the NSCLC dataset based on the public single-cell RNAseq (scRNAseq) dataset. Figure 2A The significantly increased serum protein gene expression levels identified in each of the cell types shown indicate that most proteins are of bone marrow origin, including NGAL (LCN2), TRFL (LTF), LCAT, VCAM1, APOC4, LYAM1 (SELL), CD5L, MARCO, CAMP, APOE, APOC2, CD163, LYSC (LYZ), APOA2, PERM (MPO), CSF1R, CD44, and B2MG (B2M). For protein-gene pairs with different names, the gene names are shown in italics in parentheses.

[0120] Figure 2C This is a set of KM curves showing progression-free survival (PFS) for patients using combinations of all significantly increased proteins (MARCO, CAMP, CD163, CSF1R, CD5L, NGAL (LCN2), GAPR1, APOC1, APOC2, APOC3, and APOC4) relative to those with low (dashed) or high (solid) serum myeloid protein levels at C2D1 on day 1 of cycle 1 (C1D1), determined by the median combined fractional cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0121] Figure 2DThis is a set of KM curves showing overall survival (OS) for patients with low (dashed line) or high (solid line) serum myeloid protein levels at C2D1 using combinations of all significantly increased proteins (MARCO, CAMP, CD163, CSF1R, CD5L, NGAL (LCN2), GAPR1, APOC1, APOC2, APOC3, and APOC4) at C1D1, determined by the median combined fractional cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0122] Figure 2E This is a scatter plot showing the levels of soluble CD163 (sCD163) detected by enzyme-linked immunosorbent assay (ELISA) versus those detected by mass spectrometry (Biognosys PQ500). TM The correlation between the detected CD163 levels.

[0123] Figure 2F This is a set of KM curves showing progression-free survival (PFS) in patients with low (dashed) or high (solid) fold changes in sCD163, determined by the median fold change cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0124] Figure 2G This is a set of KM curves showing overall survival (OS) for patients with low (dashed) or high (solid) fold changes in sCD163, determined by the median fold change cutoff. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0125] Figure 3A This is a unified manifold approximation and projection (UMAP) that displays single peripheral blood mononuclear cells (PBMCs) stained by cell type from patients treated with tirelinumab + atezolizumab combination therapy (n = 407,219). ILC: innate lymphocytes; MDSC: bone marrow-derived suppressor cells.

[0126] Figure 3B This is a box plot showing the proportion of proliferating cells in PBMCs at day 1 of cycle 1 (C1D1), day 15 of cycle 1 (C1D15), day 1 of cycle 2 (C2D1), and day 1 of cycle 4 (C4D1) of the tireliumab + atezolizumab combination therapy. The box plot shows the center line (median), the box (interquartile range; IQR; the range between the 25th and 75th percentiles), and the whiskers (1.58 × IQR). Means at each time point are connected by solid black lines. Samples from the same patient at different time points are connected by gray lines. The p-values ​​shown are calculated using a paired two-tailed Student's t-test and BH adjustment.

[0127] Figure 3CThis is a box plot showing the proportion of CD4+ T cells as Tregs at C1D1, C1D15, C2D1, and C4D1 of tisrelinumab plus atezolizumab combination therapy. Box plot center line, median; box, quartile range; whiskers, 1.58 × IQR. Means at each time point are connected by solid black lines. Samples from the same patient at different time points are connected by gray lines. The p-values ​​shown were calculated using a paired two-tailed Student's t-test and BH adjustment.

[0128] Figure 3D This is a set of box plots showing the proportion of total monocytes as classic monocytes (left) or intermediate monocytes (right) at C1D1, C1D15, C2D1, and C4D1 of the tisrelimumab + atezolizumab combination therapy. Box plot center line, median; box, quartile range; whiskers, 1.58 × IQR. Means at each time point are connected by solid black lines. Samples from the same patient at different time points are connected by gray lines. P-values ​​shown were calculated using a paired two-tailed Student's t-test and adjusted for BH.

[0129] Figure 3E This is a set of heatmaps showing the levels of the pathways shown in the indicated immune cell types in samples obtained during treatment (C1D15, C2D1, C4D1) from patients with NSCLC (n=15 pairs) compared to those obtained at baseline (C1D1). The hue represents the significance of the false discovery rate (FDR). Red indicates enrichment in samples during treatment, and blue indicates enrichment in baseline samples. P-values ​​were calculated using a nonparametric permutation test, and a black asterisk indicates FDR < 0.05. TNFA, tumor necrosis factor α; TGF, transforming growth factor; NFKB, nuclear factor κB.

[0130] Figure 4A This is a set of growth curves showing the tumor volume (mm²) over time in BALB / c mice implanted with syngeneic CT26 tumors. 3 Tumor cells were allowed to grow for two weeks, and then treated with control IgG2a, anti-PD-L1 and / or anti-T cell immunoglobulin and ITIM domain (anti-TIGIT) mIgG2a-LALAPG (fraction crystallizable region (Fc) - inactive), mIgG2b, or mIgG2a. Data represent one independent experiment per group of n = 10 mice.

[0131] Figure 4BThis is a set of plots showing the mean fluorescence intensity (MFI) of major histocompatibility complex II (MHC-II) on the cell surface of tumor-infiltrating dendritic cells (DCs), macrophages, and monocytes, along with a set or representative histograms associated with monocyte data. DCs: *, P = 0.0264; **, P = 0.0043. Macrophages: *, P = 0.0119. Monocytes: **, P = 0.0026; **, P = 0.0017. Mean + / - SEM, using one-way ANOVA and Dunnett multiple comparisons, with the anti-PD-L1 monotherapy group designated as the control. Each point represents data from one mouse, and n = 5 per group.

[0132] Figure 4C The figure shows the proportions of interferon-γ (IFNg)+ and TNFα+ tumor-infiltrating CD8+ T cells after the treatment shown, along with a pair of representative fluorescence-activated cell sorting (FACS) plots illustrating the gating strategies used to identify these cells. *, P = 0.0007. Mean + / - SEM, using one-way ANOVA and Dunnett multiple comparisons, with the anti-PD-L1 monotherapy group designated as the control.

[0133] Figure 4D The figure shows the proportions of IFNg+ and TNFa+ infiltrating non-Treg (FoxP3-)CD4+ T cells after the indicated treatment, along with a pair of representative fluorescence-activated cell sorting (FACS) plots illustrating the gating strategies used to identify these cells. *P = 0.0163, ****P < 0.0001. Mean + / - SEM, using one-way ANOVA and Dunnett multiple comparisons, with the anti-PD-L1 monotherapy group designated as the control.

[0134] Figure 4E This is a set of figures showing the proportions of total CD45+ cells as FoxP3-non-Treg CD4+ T cells (left), FoxP3+Treg CD4+ T cells (middle), or CD8+ T cells (right) after the treatments shown. *P = 0.0115. Mean + / - SEM, using one-way ANOVA and Dunnett multiple comparisons, with the anti-PD-L1 monotherapy group designated as the control group.

[0135] Figure 4F The figure shows the ratio of CD8+ T cells to FoxP3+Treg CD4+ T cells after the treatment shown. Mean + / - SEM, using one-way ANOVA and Dunnett multiple comparisons, with the anti-PD-L1 monotherapy group designated as the control group.

[0136] Figure 5AThis is a pair of UMAPs showing tumor-infiltrating lymphocytes (top) and bone marrow cells (bottom) stained by cell type from BALB / c mice.

[0137] Figure 5B It is a pair of bubble charts, which show, as Figure 5A The expression of marker genes shown is illustrated in tumor-infiltrating T cells and NK cells (left) and bone marrow cells (right). A broken y-axis was used to make the y-axis ranges comparable and to better compare between treatments. P-values ​​were calculated using the Wilcoxon rank-sum test.

[0138] Figure 5C The left image shows a bubble plot illustrating the expression of major histocompatibility complex (MHC) genes in all tumor-infiltrating monocytes and macrophages in the indicated treatments, while the right image shows gene expression in monocytes and macrophages grouped after treatment with anti-PD-L1 + anti-TIGIT IgG2b versus anti-PD-L1 (middle) or anti-PD-L1 + anti-TIGIT IgG2a versus anti-PD-L1 (right). A broken y-axis was used to make the y-axis ranges comparable and to better compare between treatments. P-values ​​were calculated using the Wilcoxon rank-sum test.

[0139] Figure 5D The left image shows a bubble plot illustrating the expression of memory-like and exhaustion genes in total tumor-infiltrating CD8+ T cells (combination) across the treatments shown, along with a pair of volcano plots (middle and right) showing gene expression in CD8+ T cells after treatment with anti-PD-L1 + anti-TIGIT IgG2b versus anti-PD-L1 (middle) or anti-PD-L1 + anti-TIGIT IgG2a versus anti-PD-L1 (right). A broken y-axis was used to make the y-axis ranges comparable and to better compare between treatments. P-values ​​were calculated using the Wilcoxon rank-sum test.

[0140] Figure 5E The left image shows a bubble plot illustrating the expression of immunosuppressive genes in tumor-infiltrating CD4 Tregs during the indicated treatments, while the right image shows a pair of volcano plots illustrating gene expression in CD4 Tregs after treatment with anti-PD-L1 + anti-TIGIT IgG2b with anti-PD-L1 (middle) or anti-PD-L1 + anti-TIGIT IgG2a with anti-PD-L1 (right). P-values ​​were calculated using the Wilcoxon rank-sum test.

[0141] Figure 6A This is UMAP, which displays peripheral blood cells stained by cell type.

[0142] Figure 6B It is a bubble chart, which displays as follows: Figure 6A The expression of the marker genes shown in the cell types shown.

[0143] Figure 6C The image is a heatmap showing scaled gene expression of marker genes that distinguish classical, non-classical, and intermediate monocytes (top), and the expression pattern of the FCγ receptor (FcγR) in the monocyte subset shown (bottom).

[0144] Figure 6D This is a set of volcano plots showing gene expression in classical (left), intermediate (middle), and non-classical (right) monocytes treated with anti-PD-L1 + anti-TIGIT-IgG2a and anti-PD-L1. P-values ​​were calculated using the Wilcoxon rank-sum test.

[0145] Figure 7A This is a pair of forest plots showing the relationship between high or low expression of the gene shown in the tumor and PFS (left) or OS (right) in patients treated with tireliumab + atezolizumab and placebo + atezolizumab. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0146] Figure 7B These are a set of Kaplan-Meier curves comparing progression-free survival (PFS) between patients with tumors enriched with tumors (TAM) and those with tumors not enriched with TAM from the phase 3 NSCLC OAK study of patients who received atezolizumab monotherapy. Patients were divided into two groups based on median eigenvalue scores. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0147] Figure 7C These are a set of Kaplan-Meier curves comparing overall survival (OS) between patients with tumors enriched with tumors (TAM) and those with tumors not enriched with TAM from the phase 3 NSCLC OAK study of patients who received atezolizumab monotherapy. Patients were divided into two groups based on median eigenvalue scores. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0148] Figure 7D These are a set of Kaplan-Meier curves comparing progression-free survival (PFS) between patients with Treg-enriched tumors and those with non-Treg-enriched tumors from the phase 3 NSCLC OAK study who received atezolizumab monotherapy. Patients were divided into two groups based on median eigenvalue scores. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0149] Figure 7EThis is a set of Kaplan-Meier curves comparing overall survival (OS) between patients with Treg-enriched tumors and those with non-Treg-enriched tumors from the phase 3 NSCLC OAK study who received atezolizumab monotherapy. Patients were divided into two groups based on median eigenvalue scores. Hazard ratios and 95% confidence intervals were determined using a univariate Cox model.

[0150] Figure 8A This is a scatter plot showing the correlation between TAM gene trait scores and the proportion of CD68+ cells in the total cell count, as quantified by mIF. Two-tailed Pearson correlation.

[0151] Figure 8B This is a scatter plot showing the correlation between Treg gene eigenvalues ​​and the proportion of total cells that are FoxP3+, as quantified by mIF. Two-tailed Pearson correlation.

[0152] Figure 9A This is a scatter plot showing the S and G2M cell cycle phase fractions of individual cells. Cells identified as being in a proliferative or non-proliferative state are identified by color.

[0153] Figure 9B It is a bar chart that shows the proportion of proliferating cells classified as belonging to each of the cell types shown.

[0154] Figure 9C This is a set of box-and-whisker plots showing the proportions of proliferating cells as CD4+ non-naïve T cells, CD8+ non-naïve T cells, and NK cells at C1D1, C1D15, C2D1, and C4D1 of tirelimumab + atezolizumab combination therapy. Box plot center line, median; box, quartile range; whiskers, 1.58 × IQR. Means at each time point are connected by solid black lines. Samples from the same patient at different time points are connected by gray lines. P-values ​​shown were calculated using a paired two-tailed Student's t-test and BH adjustment.

[0155] Figure 9D This is a set of box-and-whisker plots showing the proportion of PBMCs identified as belonging to the indicated cell types at C1D1, C1D15, C2D1, and C4D1 of the tisrelimumab + atezolizumab combination therapy. Box plot center line, median; box, quartile range; whiskers, 1.58 × IQR. Means at each time point are connected by solid black lines. Samples from the same patient at different time points are connected by gray lines. P-values ​​shown were calculated using a paired two-tailed Student's t-test and BH adjustment.

[0156] Figure 9EThis is a set of box-and-whisker plots showing the proportion of PBMCs identified as belonging to the indicated cell type at C1D1, C1D15, C2D1, and C4D1 in responders (complete or partial response (CRPR)) and non-responders (stable or progressive disease (SDPD)) receiving the tireliumab + atezolizumab combination therapy. Box plot center line, median; box, quartile range; whiskers, 1.58 × IQR. Means at each time point are connected by solid black lines. Samples from the same patient at different time points are connected by gray lines. Nominal p-values ​​derived from unpaired two-tailed Student's t-tests are shown, and red asterisks represent significance levels where *p < 0.05.

[0157] Figure 10A This is a pair of UMAPs showing tumor-infiltrating T cells and NK cells (top) and bone marrow cells (bottom) stained by cell type from BALB / c mice.

[0158] Figure 10B It is a pair of bubble charts, which show, as Figure 10A The expression of marker genes shown in T cells and NK cells (left) and bone marrow cells (right).

[0159] Figure 10C It is a bubble chart showing the scaled expression of MHC and cytokine genes in tumor macrophages and monocytes in the treatments shown.

[0160] Figure 10D It is a bubble chart showing the scaled expression of memory-like genes and exhaustion genes in total tumor CD8+ T cells under the indicated treatment.

[0161] Figure 10E It is a bubble chart that shows the scaled expression of the immunosuppressive genes shown in the tumor CD4+Treg in the treatment shown.

[0162] Figure 10F This is UMAP, which displays single peripheral blood cells stained by cell type.

[0163] Figure 10G It is a bubble chart, which displays as follows: Figure 10F The expression of the marker genes shown in the cell types shown.

[0164] Figure 10H The image is a heatmap showing scaled gene expression of marker genes that distinguish classical, non-classical, and intermediate monocytes (top), and the expression pattern of FcγR in the monocyte subset shown (bottom).

[0165] Figure 10IIt is a bubble chart showing scaled expression of MHC and interferon response genes as shown in non-classical monocytes from the treatment group indicated.

[0166] Figure 11A This is a set of growth curves showing the tumor volume (mm²) over time in BALB / c mice implanted with syngeneic CT26 tumors. 3 Tumor cells were allowed to grow for two weeks, then treated with control IgG2a and / or anti-TIGIT mIgG2a-LALAPG, mIgG2b, or mIgG2a. Data represent an independent experiment.

[0167] Figure 11B This is a set of growth curves showing the tumor volume (mm²) over time in wild-type (top) or FcγR knockout (bottom) BALB / c mice implanted with syngeneic CT26 tumors. 3 Tumor cells were allowed to grow for two weeks, then treated with either control IgG2a or anti-PD-L1 and anti-TIGIT mIgG2a. Data represent one independent experiment.

[0168] Figure 12A This is a pair of graphs showing the proportions of TCF1+ and SLAMF6+ (memory-like) Tp70+CD226+ T cells in mice carrying CT26 tumors after treatment with control antibody or anti-PD-L1 plus anti-TIGIT mIgG2a-LALAPG or mIgG2a antibody. Statistics are from one-way ANOVA and Tukey multiple comparisons. *, p<0.05; **, p<0.01; ****, p<0.0001.

[0169] Figure 12B These are two graphs showing the proportion of Tox+ (terminally differentiated effector T cells) gp70+CD226+ T cells in mice carrying CT26 tumors after treatment with control antibody or anti-PD-L1 plus anti-TIGIT mIgG2a-LALAPG or mIgG2a antibody. Statistics are from one-way ANOVA and Tukey multiple comparisons. *, p<0.05; **, p<0.01; ****, p<0.0001.

[0170] Figure 13AThis is a set of box plots showing the proportion of total PBMCs belonging to the indicated cell types in mice treated with IgG2a isotype control (B1); aPD-L1 (B2); aTIGIT-IgG2b (B3); aTIGIT-IgG2a (B4); aPD-L1+aTIGIT-IgG2b (B5); or aPD-L1+aTIGIT-IgG2a (B6). Center line, median; box, quartile range; whiskers, 1.58 × IQR. Nominal p-values ​​for unpaired two-tailed Student's t-tests are shown in gray; p-values ​​adjusted by Dunnett's multiple comparisons are shown in black.

[0171] Figure 13B This is a set of volcano plots showing gene expression in classical (left), intermediate (middle), and non-classical (right) monocytes from mice treated with anti-PD-L1 + anti-TIGIT-IgG2b and anti-PD-L1. P-values ​​were calculated using the Wilcoxon rank-sum test.

[0172] Figure 14A This is a volcano plot showing the relative expression levels of the genes shown in the diagram from tumor-infiltrating CD8+ T cells of BALB / c mice implanted with syngeneic CT26 tumors and treated with Fc-enabled anti-TIGIT IgG2a and anti-PD-L1 antibodies, compared to mice treated with anti-PD-L1 antibodies alone. T-effect memory genes (“memory”) and exhaustion-related genes (“exhaustion”) are indicated by color. FC: Fold change relative to anti-PD-L1 monotherapy.

[0173] Figure 14B This is a heatmap showing the mean expression level (indicated by dot color) and the percentage of cells expressing the indicated genes (indicated by dot size) from tumor-infiltrating CD8+ T cells of BALB / c mice implanted with homologous CT26 tumors, in which the indicated genes were treated with control IgG2a (T1); anti-PD-L1 antibody (T2); mIgG2b anti-TIGIT antibody (T3); mIgG2a anti-TIGIT antibody (T4); anti-PD-L1 antibody and mIgG2b anti-TIGIT antibody (T5); or anti-PD-L1 antibody and mIgG2a anti-TIGIT antibody (T6). Yellow indicates high expression levels; blue indicates low expression levels. The color indicates the scaled mean expression (i.e., the mean gene expression of cells in the group), where the mean of the scaled expression is 0 and the standard deviation (SD) is 1.

[0174] Figure 14CThis is a volcano plot, showing the relative expression levels of the genes shown in tumor-infiltrating CD4+ T cells (Tregs) from BALB / c mice implanted with syngeneic CT26 tumors and treated with Fc-enabled anti-TIGIT IgG2a and anti-PD-L1 antibodies, compared to mice treated with anti-PD-L1 antibodies alone. Immunosuppressive genes are indicated by color. FC: Fold change relative to anti-PD-L1 monotherapy.

[0175] Figure 14D This is a heatmap showing the mean expression level (indicated by dot color) and the percentage of cells expressing the indicated gene (indicated by dot size) from tumor-infiltrating CD4+ T cells (Tregs) of BALB / c mice implanted with homologous CT26 tumors, treated with control IgG2a (T1); anti-PD-L1 antibody (T2); mIgG2b anti-TIGIT antibody (T3); mIgG2a anti-TIGIT antibody (T4); anti-PD-L1 antibody and mIgG2b anti-TIGIT antibody (T5); or anti-PD-L1 antibody and mIgG2a anti-TIGIT antibody (T6). Yellow indicates high expression level; blue indicates low expression level. The color indicates the scaled mean expression (i.e., the mean gene expression of cells in the group), where the mean of the scaled expression is 0 and SD is 1.

[0176] Figure 14E This is a volcano plot showing the relative expression levels of the genes shown in the diagram from tumor-infiltrating mononuclear cells of BALB / c mice implanted with syngeneic CT26 tumors and treated with Fc-enabled anti-TIGIT IgG2a and anti-PD-L1 antibodies, compared to mice treated with anti-PD-L1 antibodies alone. FC: Fold change relative to anti-PD-L1 monotherapy. MHC-related genes (“MHC”) and other genes (“Others”) are indicated by color.

[0177] Figure 14F This is a heatmap showing the mean expression level (indicated by dot color) and the percentage of cells expressing the indicated genes (indicated by dot size) from tumor-infiltrating mononuclear cells of BALB / c mice implanted with homologous CT26 tumors, treated with control IgG2a (T1); anti-PD-L1 antibody (T2); mIgG2b anti-TIGIT antibody (T3); mIgG2a anti-TIGIT antibody (T4); anti-PD-L1 antibody and mIgG2b anti-TIGIT antibody (T5); or anti-PD-L1 antibody and mIgG2a anti-TIGIT antibody (T6). Yellow indicates high expression levels; blue indicates low expression levels. The color indicates the scaled mean expression (i.e., the mean gene expression of cells in the group), where the mean of the scaled expression is 0 and SD is 1. Detailed Implementation

[0178] I. Overview

[0179] This invention is based, at least in part, on the surprising finding that for tirelimumab plus atezolizumab combination therapy, higher abundance of immunosuppressive cells, particularly tumor-associated macrophages (TAMs) and regulatory T cells (Tregs), was associated with improved objective response rate (ORR), overall survival (OS), and progression-free survival (PFS), but not for atezolizumab monotherapy. Specifically, analysis of gene expression in tumor samples from patients in the phase 2 CITYSCAPE study (GO30103) revealed that above-median TAM and Treg gene signatures were each associated with improved outcomes from tirelimumab plus atezolizumab combination therapy. Furthermore, analysis of serum samples collected from CITYSCAPE patients before and during treatment (Day 1 of Cycle 2 (C2D1) and Day 1 of Cycle 3 (C3D1)) revealed a statistically significant increase in bone marrow-related protein peptides in the tirelinumab + atezolizumab combination therapy arm, such as MARCO (macrophage receptor with collagen structure), CSF1R, CD163, cAMP, CD5L, and apolipoproteins (APOC2 / 3 / 4), indicating that bone marrow activation is a treatment-specific effect. It was also surprisingly found that for patients with serum myeloid protein increases >median, the increase in these myeloid protein levels was associated with longer progression-free survival (PFS) and overall survival (OS) in patients receiving the tirelinumab + atezolizumab combination therapy compared to patients receiving atezolizumab monotherapy. Therefore, the combination therapy showed a transient increase in serum myeloid protein in the arm associated with the difference in improvement in PFS and OS, which is related to the combination therapy of tirelinumab + atezolizumab, suggesting that myeloid cells may play a key role in the enhanced antitumor efficacy of tirelinumab + atezolizumab.

[0180] This article also discovered novel upregulated pathways in monocytes that have not been reported in atezolizumab monotherapy and are specific to tireliumab + atezolizumab combination therapy, including the MYC-targeting pathway, which has been shown to regulate macrophage polarization.

[0181] It has also been surprisingly discovered that the interaction between the tirelinumab Fc domain and the Fcγ receptor is essential for the observed bone marrow activation.

[0182] II. General Techniques and Definitions

[0183] Those skilled in the art will readily understand and generally use conventional methods to employ the techniques and procedures described or referenced herein, such as, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual 3rd Edition (2001), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (edited by F.M. Usubel et al., (2003)); Methods in Enzymology series (Academic Press, Inc.): PCR 2: A Practical Approach (edited by M.J. MacPherson, B.D. Hames, and G.R. Taylor, (1995)); Harlow and Lane (edited by Harlow and Lane, (1988); Antibodies, A Laboratory Manual, and Animal Cell Culture (edited by R.R. Freshney, (1987)); Oligonucleotide Synthesis (edited by M.J. Gait, 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Manual. Notebook (edited by J.E.C. Elis, 1998) Academic Press; Animal Cell Culture (edited by R.R. Freshney, 1987); Introduction to Cell and Tissue Culture (edited by J.M. Pather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (edited by A. Doyle, J.B. Griffiths, and D.G. Newell, 1993-8) J. Wiley and Sons; Handbook of Experimental Immunology (edited by D.M. Weir and C.C. Blackwell); Gene Transfer Vectors for Mammalian Cells (edited by J.M. Miller and M.P. Roberts, 1998)Calos, 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 (CAJaneway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (D. Catty, ed., IRL Press, 1988-1989); Monoclonal Antibodies: A Practical Approach (P. Shepherd and C. Dean, ed., Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane, Cold Spring Harbor Laboratory). The widely used methods described in *The Antibodies* (M. Zanetti and JD Capra, eds., Harwood Academic Publishers, 1999), and *Cancer: Principles and Practice of Oncology* (VT DeVita et al., eds., JBLippincott Company, 1993) are as follows.

[0184] It should be understood that the aspects and embodiments of the invention described herein include those referred to by "comprising," "consisting of," and "substantially consisting of." Unless otherwise indicated, as used herein, the singular forms "a / an" and "the / described" include the plural of the referred to items.

[0185] As used herein, the term "about" refers to a common range of error for a corresponding value that is readily known to those skilled in the art. References to "about" values ​​or parameters herein include (and describe) embodiments relating to that value or parameter itself. For example, a description referring to "about X" includes a description of "X".

[0186] The terms “amount,” “level,” or “expression level,” used interchangeably herein for biomarkers, refer to the detectable level in a biological sample. “Expression” generally refers to the process by which information (e.g., gene-encoded and / or epigenetic) is converted into structures present and functioning within the cell. Therefore, as used herein, “expression” can refer to transcription into a polynucleotide, translation into a polypeptide, or even polynucleotide and / or polypeptide modification (e.g., post-translational modification of a polypeptide). Fragments of transcribed polynucleotides, translated polypeptides, or polynucleotide and / or polypeptide modifications (e.g., post-translational modifications of a polypeptide) should also be considered expressed, regardless of whether they originate from transcripts generated through alternative splicing or degradation, or from post-translational processing of polypeptides (e.g., through proteolysis). “Expressed genes” include those transcribed into polynucleotides such as mRNA and then translated into polypeptides, as well as those transcribed into RNA but not translated into polypeptides (e.g., transfer RNA and ribosomal RNA). Expression levels can be measured using methods known to those skilled in the art and disclosed herein.

[0187] The terms "detecting" (or "detection") are used in the broadest sense herein to encompass both qualitative and quantitative measurements of target molecules. Detection includes identifying the presence of target molecules in a sample and determining whether the target molecules are present in the sample at detectable levels. Detection can be direct or indirect.

[0188] The presence and / or expression levels / amounts of the various biomarkers described herein in a sample can be analyzed by a variety of methods, many of which are known in the art and understood by those skilled in the art, including but not limited to immunohistochemistry (“IHC”), Western blot analysis, immunoprecipitation, molecular binding assays, ELISA, ELIFA, fluorescence activated cell sorting (“FACS”), MassARRAY, proteomics, blood-based quantitative assays (e.g., serum ELISA), biochemical enzyme activity assays, in situ hybridization, fluorescence in situ hybridization (FISH), DNA blot analysis, RNA blot analysis, whole genome sequencing, and massively parallel DNA sequencing (e.g., next-generation sequencing). Polymerase chain reaction (PCR) (including quantitative real-time PCR (qRT-PCR) and other amplification-type detection methods such as branched DNA, SISBA, TMA), RNA-seq, microarray analysis, gene expression profiling and / or serial analysis of gene expression (“SAGE”), and any of a variety of assays that can be performed by protein, gene and / or tissue analysis. Typical protocols for evaluating the state of genes and gene products can be found, for example, in Ausubel et al., ed., *Current Protocols in Molecular Biology*, 1995, Units 2 (RNA blotting), 4 (DNA blotting), 15 (immunoblotting), and 18 (PCR analysis). Multiplex immunoassays, such as those available from Rules Based Medicine or Meso Scale Discovery (“MSD”), can also be used.

[0189] Unless otherwise specified, the term "complement C1q subunit C" or "C1QC" as used herein broadly refers to any naturally occurring C1QC from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length C1QC and dissociated regions or domains of C1QC, such as C1QCECD. The term also encompasses naturally occurring variants of C1QC, such as splice variants or allelic variants. An exemplary amino acid sequence of human C1QC is shown under UniProt accession number P02747. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions in C1QC that do not affect C1QC function and / or activity.

[0190] Unless otherwise specified, the terms “macrophage scavenger receptor type I and II” or “MSR1” as used herein broadly refer to any naturally occurring MSR1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length MSR1 and isolated regions or domains of MSR1, such as MSR1ECD. The term also encompasses naturally occurring variants of MSR1, such as splice variants or allelic variants. An exemplary amino acid sequence of human MSR1 is shown under UniProt accession number P21757. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of MSR1 that do not affect MSR1 function and / or activity.

[0191] Unless otherwise specified, the terms “macrophage mannose receptor 1” or “MRC1” as used herein broadly refer to any naturally occurring MRC1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length MRC1 and isolated regions or domains of MRC1, such as MRC1ECD. The term also encompasses naturally occurring variants of MRC1, such as splice variants or allelic variants. An exemplary amino acid sequence of human MRC1 is shown under UniProt accession number P22897. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions in MRC1 that do not affect MRC1 function and / or activity.

[0192] Unless otherwise specified, the terms “V-set and immunoglobulin domain-containing protein 4” or “VSIG4” as used herein broadly refer to any naturally occurring VSIG4 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length VSIG4 and isolated regions or domains of VSIG4, such as VSIG4 ECD. The term also encompasses naturally occurring variants of VSIG4, such as splice variants or allelic variants. An exemplary amino acid sequence of human VSIG4 is shown under UniProt accession number Q9Y279. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of VSIG4 that do not affect VSIG4 function and / or activity.

[0193] Unless otherwise specified, the term "secreted phosphoprotein 1" or "SPP1" as used herein refers to any naturally occurring SPP1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length SPP1 and isolated regions or domains of SPP1, such as SPP1 ECD. The term also encompasses naturally occurring variants of SPP1, such as splice variants or allelic variants. An exemplary amino acid sequence of human SPP1 is shown under UniProt accession number P10451. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in SPP1 that do not affect SPP1 function and / or activity.

[0194] Unless otherwise specified, the terms "macrophage receptor with collagen structure" or "MARCO" as used herein refer to any naturally occurring MARCO from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses both full-length MARCOs and separated regions or domains of MARCOs, such as MARCO ECDs. The term also encompasses naturally occurring variants of MARCOs, such as splice variants or allelic variants. An exemplary amino acid sequence of a human MARCO is shown under UniProt accession number Q9UEW3. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in MARCOs that do not affect MARCO function and / or activity.

[0195] Unless otherwise specified, the terms “tartrate-resistant acid phosphatase type 5” or “ACP5” as used herein broadly refer to any naturally occurring ACP5 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length ACP5 and isolated regions or domains of ACP5, such as ACP5 ECD. The term also encompasses naturally occurring variants of ACP5, such as splice variants or allelic variants. An exemplary amino acid sequence of human ACP5 is shown under UniProt accession number P13686. The invention also considers minor sequence variations, particularly conserved amino acid substitutions of ACP5 that do not affect ACP5 function and / or activity.

[0196] Unless otherwise specified, the terms “mast cell-expressed membrane protein 1” or “MCEMP1” as used herein broadly refer to any naturally occurring MCEMP1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length MCEMP1 and isolated regions or domains of MCEMP1, such as MCEMP1 ECD. The term also encompasses naturally occurring variants of MCEMP1, such as splice variants or allelic variants. An exemplary amino acid sequence of human MCEMP1 is shown under UniProt accession number Q8IX19. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of MCEMP1 that do not affect the function and / or activity of MCEMP1.

[0197] Unless otherwise specified, the terms “sterol 27-hydroxylase” or “CYP27A1” as used herein refer to any naturally occurring CYP27A1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length CYP27A1 and isolated regions or domains of CYP27A1, such as CYP27A1 ECD. The term also encompasses naturally occurring variants of CYP27A1, such as splice variants or allelic variants. An exemplary amino acid sequence of human CYP27A1 is shown under UniProt accession number Q02318. The invention also considers minor sequence variations, particularly conserved amino acid substitutions of CYP27A1 that do not affect the function and / or activity of CYP27A1.

[0198] Unless otherwise specified, the terms “oxidized low-density lipoprotein receptor 1” or “OLR1” as used herein broadly refer to any naturally occurring OLR1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length OLR1 and isolated regions or domains of OLR1, such as OLR1 ECD. The term also encompasses naturally occurring variants of OLR1, such as splice variants or allelic variants. An exemplary amino acid sequence of human OLR1 is shown under UniProt accession number P78380. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of OLR1 that do not affect OLR1 function and / or activity.

[0199] Unless otherwise specified, the terms "granule protein precursor" or "GRN" as used herein refer to any naturally occurring GRN from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses both full-length GRNs and separated regions or domains of GRNs, such as GRN ECDs. The term also encompasses naturally occurring variants of GRNs, such as splice variants or allelic variants. An exemplary amino acid sequence of a human GRN is shown under UniProt accession number P28799. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in GRNs that do not affect GRN function and / or activity.

[0200] Unless otherwise specified, the terms “glioma pathogenesis-associated protein 2” or “GLIPR2” as used herein broadly refer to any naturally occurring GLIPR2 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length GLIPR2 and isolated regions or domains of GLIPR2, such as GLIPR2ECD. The term also encompasses naturally occurring variants of GLIPR2, such as splice variants or allelic variants. An exemplary amino acid sequence of human GLIPR2 is shown under UniProt accession number Q9H4G4. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of GLIPR2 that do not affect GLIPR2 function and / or activity.

[0201] Unless otherwise specified, the terms "protein 4 containing a repressor protein domain" or "ARRDC4" as used herein broadly refer to any naturally occurring ARRDC4 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length ARRDC4 and isolated regions or domains of ARRDC4, such as ARRDC4 ECD. The term also encompasses naturally occurring variants of ARRDC4, such as splice variants or allelic variants. An exemplary amino acid sequence of human ARRDC4 is shown under UniProt accession number Q8NCT1. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions in ARRDC4 that do not affect ARRDC4 function and / or activity.

[0202] Unless otherwise specified, the term "apolipoprotein E" or "APOE" as used herein refers to any naturally occurring APOE from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length APOE and isolated regions or domains of APOE, such as APOE ECD. The term also encompasses naturally occurring variants of APOE, such as splice variants or allelic variants. An exemplary amino acid sequence of human APOE is shown under UniProt accession number P02649. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in APOE that do not affect APOE function and / or activity.

[0203] Unless otherwise specified, the terms “folate receptor β” or “FOLR2” as used herein refer to any naturally occurring FOLR2 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length FOLR2 and dissociated regions or domains of FOLR2, such as FOLR2 ECD. The term also encompasses naturally occurring variants of FOLR2, such as splice variants or allelic variants. An exemplary amino acid sequence of human FOLR2 is shown under UniProt accession number P14207. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in FOLR2 that do not affect FOLR2 function and / or activity.

[0204] Unless otherwise specified, the terms “cathepsin D” or “CTSD” as used herein refer to any naturally occurring CTSD from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length CTSDs and separated regions or domains of CTSDs, such as CTSD ECDs. The term also encompasses naturally occurring variants of CTSDs, such as splice variants or allelic variants. An exemplary amino acid sequence of human CTSD is shown under UniProt accession number P07339. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in CTSDs that do not affect CTSD function and / or activity.

[0205] Unless otherwise specified, the terms “cathepsin inhibitor antimicrobial peptide” or “CAMP” as used herein refer to any naturally occurring CAMP from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses both full-length CAMP and separated regions or domains of CAMP, such as CAMP ECD. The term also encompasses naturally occurring variants of CAMP, such as splice variants or allelic variants. An exemplary amino acid sequence of human CAMP is shown under UniProt accession number P49913. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in CAMP that do not affect CAMP function and / or activity.

[0206] Unless otherwise specified, the terms “CD5 antigen-like” or “CD5L” as used herein refer to any naturally occurring CD5L from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length CD5L and dissociated regions or domains of CD5L, such as CD5L ECD. The term also encompasses naturally occurring variants of CD5L, such as splice variants or allelic variants. An exemplary amino acid sequence of human CD5L is shown under UniProt accession number O43866. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of CD5L that do not affect CD5L function and / or activity.

[0207] Unless otherwise specified, the terms “scavenger receptor cysteine-rich type 1 protein M130” or “CD163” as used herein broadly refer to any naturally occurring CD163 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length CD163 and dissociated regions or domains of CD163, such as CD163 ECD. The term also encompasses naturally occurring variants of CD163, such as splice variants or allelic variants. An exemplary amino acid sequence of human CD163 is shown under UniProt accession number Q86VB7. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions in CD163 that do not affect CD163 function and / or activity.

[0208] Unless otherwise specified, the terms “neutrophil gelatinase-associated lipotransporter” or “NGAL” as used herein refer to any naturally occurring NGAL from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length NGALs and dissociated regions or domains of NGALs, such as NGAL ECDs. The term also encompasses naturally occurring variants of NGALs, such as splice variants or allelic variants. An exemplary amino acid sequence of human NGAL is shown under UniProt accession number P80188. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in NGALs that do not affect NGAL function and / or activity.

[0209] Unless otherwise specified, the terms “macrophage colony-stimulating factor 1 receptor” or “CSF1R” as used herein broadly refer to any naturally occurring CSF1R from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length CSF1R and dissociated regions or domains of CSF1R, such as CSF1R ECD. The term also encompasses naturally occurring variants of CSF1R, such as splice variants or allelic variants. An exemplary amino acid sequence of human CSF1R is shown under UniProt accession number P07333. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of CSF1R that do not affect CSF1R function and / or activity.

[0210] Unless otherwise specified, as used herein, the terms “CD44 antigen” or “CD44” refer to any naturally occurring CD44 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length CD44 and dissociated regions or domains of CD44, such as CD44 ECD. The term also encompasses naturally occurring variants of CD44, such as splice variants or allelic variants. An exemplary amino acid sequence of human CD44 is shown under UniProt accession number P16070. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of CD44 that do not affect CD44 function and / or activity.

[0211] Unless otherwise specified, the terms “apolipoprotein C-II” or “APOC2” as used herein refer to any naturally occurring APOC2 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length APOC2 and isolated regions or domains of APOC2, such as APOC2 ECD. The term also encompasses naturally occurring variants of APOC2, such as splice variants or allelic variants. An exemplary amino acid sequence of human APOC2 is shown under UniProt accession number P02655. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in APOC2 that do not affect APOC2 function and / or activity.

[0212] Unless otherwise specified, the terms “apolipoprotein C-III” or “APOC3” as used herein refer to any naturally occurring APOC3 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length APOC3 and isolated regions or domains of APOC3, such as APOC3 ECD. The term also encompasses naturally occurring variants of APOC3, such as splice variants or allelic variants. An exemplary amino acid sequence of human APOC3 is shown under UniProt accession number P02656. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in APOC3 that do not affect APOC3 function and / or activity.

[0213] Unless otherwise specified, the terms "apolipoprotein C-IV" or "APOC4" as used herein refer to any naturally occurring APOC4 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length APOC4 and isolated regions or domains of APOC4, such as APOC4 ECD. The term also encompasses naturally occurring variants of APOC4, such as splice variants or allelic variants. An exemplary amino acid sequence of human APOC4 is shown under UniProt accession number P55056. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in APOC4 that do not affect APOC4 function and / or activity.

[0214] Unless otherwise specified, the terms "apolipoprotein A-II" or "APOA2" as used herein refer to any naturally occurring APOA2 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length APOA2 and isolated regions or domains of APOA2, such as APOA2 ECD. The term also encompasses naturally occurring variants of APOA2, such as splice variants or allelic variants. An exemplary amino acid sequence of human APOA2 is shown under UniProt accession number P02652. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in APOA2 that do not affect APOA2 function and / or activity.

[0215] Unless otherwise specified, the terms “lactotransferrin” or “TRFL” as used herein refer to any naturally occurring TRFL from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length TRFL and separated regions or domains of TRFL, such as TRFL ECDs. The term also encompasses naturally occurring variants of TRFL, such as splice variants or allelic variants. An exemplary amino acid sequence of human TRFL is shown under UniProt accession number P02788. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in TRFL that do not affect TRFL function and / or activity.

[0216] Unless otherwise specified, the terms “vascular cell adhesion protein 1” or “VCAM1” as used herein broadly refer to any naturally occurring VCAM1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length VCAM1 and isolated regions or domains of VCAM1, such as VCAM1ECD. The term also encompasses naturally occurring variants of VCAM1, such as splice variants or allelic variants. An exemplary amino acid sequence of human VCAM1 is shown under UniProt accession number P13686. The invention also considers minor sequence variations, particularly conserved amino acid substitutions of VCAM1 that do not affect VCAM1 function and / or activity.

[0217] Unless otherwise specified, the terms “β-2-microglobulin” or “B2MG” as used herein refer to any naturally occurring B2MG from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length B2MG and isolated regions or domains of the B2MG, such as the B2MG ECD. The term also encompasses naturally occurring variants of the B2MG, such as splice variants or allelic variants. An exemplary amino acid sequence of human B2MG is shown under UniProt accession number P61769. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in the B2MG that do not affect its function and / or activity.

[0218] Unless otherwise specified, the term "forkhead box protein P3" or "FOXP3" as used herein broadly refers to any naturally occurring FOXP3 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length FOXP3 and dissociated regions or domains of FOXP3, such as FOXP3ECD. The term also encompasses naturally occurring variants of FOXP3, such as splice variants or allelic variants. An exemplary amino acid sequence of human FOXP3 is shown under UniProt accession number Q9BZS1. The invention also considers minor sequence variations, particularly conserved amino acid substitutions of FOXP3 that do not affect FOXP3 function and / or activity.

[0219] Unless otherwise specified, the terms “cytotoxic T-lymphocyte protein 4” or “CTLA4” as used herein broadly refer to any naturally occurring CTLA4 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length CTLA4 and isolated regions or domains of CTLA4, such as CTLA4 ECD. The term also encompasses naturally occurring variants of CTLA4, such as splice variants or allelic variants. An exemplary amino acid sequence of human CTLA4 is shown under UniProt accession number P16410. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions in CTLA4 that do not affect CTLA4 function and / or activity.

[0220] Unless otherwise specified, the term "interleukin-10" or "IL10" as used herein refers to any naturally occurring IL10 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length IL10 and isolated regions or domains of IL10, such as IL10 ECD. The term also encompasses naturally occurring variants of IL10, such as splice variants or allelic variants. An exemplary amino acid sequence of human IL10 is shown under UniProt accession number P22301. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of IL10 that do not affect IL10 function and / or activity.

[0221] Unless otherwise specified, the terms “tumor necrosis factor receptor superfamily member 18” or “TNFRSF18” as used herein broadly refer to any naturally occurring TNFRSF18 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length TNFRSF18 and dissociated regions or domains of TNFRSF18, such as TNFRSF18 ECD. The term also encompasses naturally occurring variants of TNFRSF18, such as splice variants or allelic variants. An exemplary amino acid sequence of human TNFRSF18 is shown under UniProt accession number Q9Y5U5. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of TNFRSF18 that do not affect the function and / or activity of TNFRSF18.

[0222] Unless otherwise specified, the terms “CC chemokine receptor type 8” or “CCR8” as used herein broadly refer to any naturally occurring CCR8 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the full-length CCR8 and dissociated regions or domains of CCR8, such as CCR8ECD. The term also encompasses naturally occurring variants of CCR8, such as splice variants or allelic variants. An exemplary amino acid sequence of human CCR8 is shown under UniProt accession number P51685. The invention also contemplates minor sequence variations, particularly conserved amino acid substitutions of CCR8 that do not affect CCR8 function and / or activity.

[0223] Unless otherwise specified, the terms “zinc finger protein Eos” or “IKZF4” as used herein broadly refer to any naturally occurring IKZF4 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length IKZF4 and isolated regions or domains of IKZF4, such as IKZF4ECD. The term also encompasses naturally occurring variants of IKZF4, such as splice variants or allelic variants. An exemplary amino acid sequence of human IKZF4 is shown under UniProt accession number Q9H2S9. The invention also considers minor sequence variations, particularly conserved amino acid substitutions of IKZF4 that do not affect the function and / or activity of IKZF4.

[0224] Unless otherwise specified, the terms “zinc finger protein Helios” or “IKZF2” as used herein broadly refer to any naturally occurring IKZF2 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses full-length IKZF2 and dissociated regions or domains of IKZF2, such as IKZF2ECD. The term also encompasses naturally occurring variants of IKZF2, such as splice variants or allelic variants. An exemplary amino acid sequence of human IKZF2 is shown under UniProt accession number Q9UKS7. The invention also considers minor sequence variations, particularly conserved amino acid substitutions in IKZF2 that do not affect IKZF2 function and / or activity.

[0225] Unless otherwise specified, the term “TIGIT” or “T-cell immune receptor with Ig and ITIM domains” as used herein refers to any natural TIGIT from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). TIGIT is also referred to in the art as DKFZp667A205, FLJ39873, V-set, and proteins 9 and V-set containing immunoglobulin domains, and proteins 3, VSIG9, VSTM3, and WUCAM containing transmembrane domains. The term encompasses “full-length”, unprocessed TIGIT (e.g., full-length human TIGIT having the amino acid sequence SEQ ID NO:30), as well as any form of TIGIT produced through cellular processing (e.g., processed human TIGIT without a signaling sequence having the amino acid sequence SEQ ID NO:31). The term also encompasses naturally occurring variants of TIGIT, such as splice variants or allelic variants. The amino acid sequence of the exemplary human TIGIT can be found with UniProt accession number Q495A1.

[0226] Unless otherwise specified, “PD-L1” or “programmed cell death ligand 1” herein refers to any naturally occurring PD-L1 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). PD-L1 is also referred to in the art as the CD274 molecule, CD274 antigen, B7 homolog 1, PDCD1 ligand 1, PDCD1LG1, PDCD1L1, B7H1, PDL1, programmed cell death ligand 1, B7-H1, and B7-H. The term also covers naturally occurring PD-L1 variants, such as splice variants or allelic variants. An exemplary amino acid sequence of human PD-L1 can be found under UniProt accession number Q9NZQ7 (SEQ ID NO:32).

[0227] The term "antagonist" is used in its broadest sense to include any molecule that partially or completely blocks, inhibits, or neutralizes the biological activity of the natural polypeptides disclosed herein. Suitable antagonist molecules specifically include 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 detectable changes in one or more biological activities typically associated with that polypeptide.

[0228] The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction between a PD-1 axis binding partner and one or more of its binding partners to eliminate T cell dysfunction caused by signal transduction along the PD-1 signaling axis, resulting in the restoration or enhancement of T cell function (e.g., proliferation, cytokine production, target cell killing). As used herein, PD-1 axis binding antagonists include PD-1 binding antagonists, PD-L1 binding antagonists, and PD-L2 binding antagonists.

[0229] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates, or interferes with signaling generated by the interaction of PD-1 with one or more binding partners (such as PD-L1, PD-L2). In some embodiments, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 with one or more binding partners. In one particular aspect, a PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and / or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies and their antigen-binding fragments, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, eliminate, or interfere with signaling generated by the interaction of PD-1 with PD-L1 and / or PD-L2. In one embodiment, a PD-1 binding antagonist may reduce negative co-stimulatory signals mediated by or through PD-1 signaling mediated by cell surface proteins expressed on T lymphocytes, thereby reducing dysfunction of dysfunctional T cells (e.g., improving effector responses to antigen recognition). In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In a specific aspect, the PD-1 binding antagonist is MDX-1106 (nivolumab). In another specific aspect, the PD-1 binding antagonist is pembrolizumab (formerly known as lambolizumab (MK-3475)) as described herein. In yet another specific aspect, the PD-1 binding antagonist is AMP-224 as described herein.

[0230] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates, or interferes with signaling generated by the interaction of PD-L1 with one or more binding partners (such as PD-1, B7-1). In some embodiments, a PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 with its binding partners. In one specific aspect, a PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, a PD-L1 binding antagonist includes an anti-PD-L1 antibody, its antigen-binding fragment, an immunoadhesin, a fusion protein, an oligopeptide, and other molecules that reduce, block, inhibit, eliminate, or interfere with signaling generated by the interaction of PD-L1 with one or more binding partners (such as PD-1, B7-1). In one embodiment, a PD-L1 binding antagonist may reduce negative co-stimulatory signals mediated by or through PD-L1 signaling mediated by cell surface proteins expressed on T lymphocytes, thereby reducing dysfunction of dysfunctional T cells (e.g., improving effector responses to antigen recognition). In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In one specific aspect, the anti-PD-L1 antibody is atezolizumab (e.g., MPDL3280A) as described herein. In another specific aspect, the anti-PD-L1 antibody is MDX-1105 as described herein. In yet another specific aspect, the anti-PD-L1 antibody is MEDI4736 as described herein.

[0231] As used herein, the term “atezolizumab” refers to an anti-PD-L1 antagonist antibody having an International Nonproprietary Name (INN) List 112 (WHO Drug Information, Vol. 28, No. 4, 2014, p. 488) or CAS Registry No. 1380723-44-3.

[0232] The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates, or interferes with signaling generated by the interaction of PD-L2 with one or more binding partners (such as PD-1). In some embodiments, a PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 with one or more binding partners. In one particular aspect, a PD-L2 binding antagonist inhibits the binding of PD-L2 to PD-1. In some embodiments, a PD-L2 antagonist includes an anti-PD-L2 antibody, its antigen-binding fragment, an immunoadhesin, a fusion protein, an oligopeptide, and other molecules that reduce, block, inhibit, eliminate, or interfere with signaling generated by the interaction of PD-L2 with one or more of its binding partners (such as PD-1). In one embodiment, a PD-L2 binding antagonist may reduce negative co-stimulatory signals mediated by or through PD-L2 signaling mediated by cell surface proteins expressed on T lymphocytes, thereby reducing dysfunction of dysfunctional T cells (e.g., improving effector responses to antigen recognition). In some embodiments, the PD-L2 binding antagonist is an immunoadhesive.

[0233] The term "anti-TIGIT antagonist antibody" refers to an antibody or its antigen-binding fragment or variant that can bind to TIGIT with sufficient affinity to substantially or completely inhibit the biological activity of TIGIT. For example, an anti-TIGIT antagonist antibody can block signaling via PVR, PVRL2, and / or PVRL3, thereby restoring functional responses (e.g., proliferation, cytokine production, target cell killing) induced by T cells from a dysfunctional state to antigen stimulation. For example, an anti-TIGIT antagonist antibody can block signaling via PVR without affecting PVR-CD226 interaction. Those skilled in the art will understand that in some cases, an anti-TIGIT antagonist antibody can antagonize one TIGIT activity without affecting another. For example, an anti-TIGIT antagonist antibody used in certain methods or uses described herein is an anti-TIGIT antagonist antibody that antagonizes TIGIT activity in response to one of the PVR interaction, PVRL3 interaction, or PVRL2 interaction, for example, without affecting or minimally affecting any other TIGIT interaction. In one embodiment, for example as measured by radioimmunoassay (RIA), the binding degree of the anti-TIGIT antagonist antibody to unrelated non-TIGIT proteins is less than about 10% of the antibody's binding degree to TIGIT. In some embodiments, the dissociation constant (K0) of the anti-TIGIT antagonist antibody bound to TIGIT is... D ) is ≤1μM, ≤100nM, ≤10nM, ≤1nM, ≤0.1nM, ≤0.01nM or ≤0.001nM (e.g., 10 -8 M or smaller, such as 10-8 M to 10 -13 M, for example, 10 -9 M to 10 -13 M). In some embodiments, the anti-TIGIT antagonist antibody binds to conserved TIGIT epitopes between TIGITs from different species or to epitopes on TIGITs that allow cross-species reactivity. In one embodiment, the anti-TIGIT antagonist antibody is tirelinumab.

[0234] As used herein, “tisrelimumab” is derived from fully human IgG1 / κMAb in open monoclonal antibody (OMT) rats, which binds to TIGIT and contains the heavy chain sequence of SEQ ID NO:33 and the light chain sequence of SEQ ID NO:34. tisrelimumab contains two N-linked glycosylation sites (N306) in its Fc domain. tisrelimumab is also described in: WHO Drug Information (International Nonproprietary Names for Medicinal Substances), Proposed INN: List 117, Volume 31, Issue 2, published on June 9, 2017 (see page 343).

[0235] As used herein, “administration” means the method of giving a subject a dose of a compound (e.g., an anti-TIGIT antagonist antibody or a PD-1 axis binding antagonist (e.g., an anti-PD-L1 antibody)) or a composition (e.g., a pharmaceutical composition, such as a pharmaceutical composition comprising an anti-TIGIT antibody and / or a PD-1 axis binding antagonist (e.g., an anti-PD-L1 antibody)). The compounds and / or compositions used in the methods described herein may be administered via, for example, intravenously (e.g., by intravenous infusion), subcutaneously, intramuscularly, intradermally, percutaneously, intra-arterially, intraperitoneally, intralesionally, intracranially, intra-articularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, locally, intratumorally, intraperitoneally, subconjunctivally, intracysticly, mucosally, intraperitoneally, intraumbilically, intraocularly, orally, locally, by inhalation, by injection, by infusion, by continuous infusion, by local perfusion of target cells, by catheter, by irrigation, as a cream, or as a lipid composition. The method of application can vary depending on a variety of factors (e.g., the compound or composition to be applied and the severity of the condition, disease, or illness to be treated).

[0236] As used herein, “systemic therapy” refers to a treatment that crosses the bloodstream and reaches multiple organ systems after a single application. The term “systemic therapy” is well known to those skilled in the art and is equivalent to systemic therapy.

[0237] The term "fixed dose" (flat dose) for a therapeutic agent (e.g., an anti-TIGIT antagonist antibody or a PD-1 axis binding antagonist (e.g., an anti-PD-L1 antibody)) in this article refers to the dose administered to the patient regardless of their weight or body surface area (BSA). Therefore, a fixed dose is not expressed in mg / kg or mg / m². 2 It is not provided in doses, but in absolute amounts as a therapeutic agent.

[0238] As used herein, the term "treatment" (or "treating") refers to a clinical intervention designed to alter the natural course of the treated individual or cells in a clinicopathological process. Ideal treatment outcomes include slowing or reducing the rate of disease progression, alleviating or reducing the severity of the disease state, and improving or alleviating prognosis. For example, an individual is successfully "treated" if one or more cancer-related symptoms are reduced or eliminated, including but not limited to reducing the proliferation (or destruction) of cancer cells, alleviating symptoms caused by the disease, improving the quality of life of a person with the disease, reducing the dosage of other medications required to treat the disease, slowing disease progression, and / or prolonging the individual's survival.

[0239] As used in this article, "in conjunction with" means applying another treatment in addition to one treatment. Thus, "in conjunction with" means applying another treatment before, during, or after applying one treatment to an individual.

[0240] "Disease" or "illness" means any condition in which treatment will benefit, including but not limited to diseases associated with some degree of abnormal cell proliferation, such as cancer, such as lung cancer, for example, non-small cell lung cancer (NSCLC).

[0241] The term "functional disorder" in immune dysfunction refers to a state of reduced immune response to antigen stimulation.

[0242] As used herein, the term “functional impairment” also includes refractory or unresponsiveness to antigen recognition, specifically, the impaired ability to translate antigen recognition into downstream T cell effector functions such as proliferation, production of cytokines (e.g., interferon gamma) and / or killing of target cells.

[0243] The terms “cancer” and “cancerous” refer to or describe a physiological condition in mammals typically characterized by uncontrolled cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More specific examples of such cancers include, but are not limited to, lung cancer, such as non-small cell lung cancer (NSCLC), which includes squamous or non-squamous NSCLC, including locally advanced unresectable NSCLC (e.g., stage IIIB NSCLC) or recurrent or metastatic NSCLC (e.g., stage IV NSCLC); lung adenocarcinoma or squamous cell carcinoma of the lung (e.g., epithelial squamous cell carcinoma of the lung (e.g., squamous cell carcinoma of the lung)); and small cell lung cancer (SCLC), which includes extensive-stage SCLC (ES-SCLC). Other examples of cancer include gastric / stomach cancer. Cancer, including gastrointestinal cancer, gastrointestinal stromal cancer, or gastroesophageal junction cancer; esophageal cancer; colon cancer; rectal cancer; colorectal cancer; peritoneal cancer; hepatocellular carcinoma; pancreatic cancer; glioblastoma; cervical cancer; ovarian cancer; liver cancer; bladder cancer (e.g., urothelial bladder cancer (UBC), muscle-invasive bladder cancer (MIBC), and BCG-refractory non-muscle-invasive bladder cancer (NMIBC)); urinary tract cancer; liver cancer; breast cancer (e.g., HER2+ breast cancer and triple-negative breast cancer (TN)). TNBC (Typoxetine-mediated nephropathy) refers to cancers that are negative for estrogen receptor (ER-), progesterone receptor (PR-), and HER2 (HER2-); endometrial cancer or uterine cancer; salivary gland cancer; kidney cancer or renal cell carcinoma (e.g., renal cell carcinoma (RCC)); prostate cancer; vulvar cancer; thyroid cancer; liver cancer; anal cancer; penile cancer; melanoma, including superficial diffuse melanoma, malignant lentigines melanoma, peripheral macular malignant melanoma, and nodular melanoma; multiple myeloma and B-cell lymphoma. (Including low-grade / follicular non-Hodgkin lymphoma (NHL); small lymphocytic (SL) NHL; intermediate / follicular NHL; intermediate diffuse NHL; high-grade immunoblastic NHL; high-grade lymphocytic NHL; high-grade small non-lytic cell NHL; giant mass NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); acute myeloid leukemia (AML); hairy cell leukemia; chronic myeloid leukemia (CML); post-transplant lymphoproliferative disorder (PTLD); and myelodysplastic syndromes (MDS), as well as abnormal angiogenesis associated with nevus hamartomatosis, edema (such as that associated with brain tumors), Meigs' syndrome, brain cancer, head and neck cancer, and related metastases.

[0244] The term "tumor" refers to all proliferative cell growth and proliferation, whether malignant or benign, as well as all precancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "proliferative disorder," "proliferative lesion," and "tumor" are not mutually exclusive in this document.

[0245] "Tumor immunity" refers to the process by which tumors evade immune recognition and clearance. Therefore, as a therapeutic concept, tumor immunity is "treated" when this evasion is mitigated and the tumor is recognized and attacked by the immune system. Examples of tumor recognition include tumor binding, tumor shrinkage, and tumor clearance.

[0246] As used in this article, “metastasis” refers to the spread of cancer from its primary site to other parts of the body. Cancer cells can detach from the primary tumor, infiltrate lymphatic vessels and blood vessels, circulate in the bloodstream, and grow in distant lesions in normal tissues elsewhere in the body (metastasis). Metastasis can be local or distant. Metastasis is a continuous process that depends on tumor cells detaching from the primary tumor, crossing the bloodstream, and stopping at a distant site. At the new site, the cells establish a blood supply and can grow to form a life-threatening mass. Excitatory and inhibitory molecular pathways within tumor cells regulate this behavior, and interactions between tumor cells and distant host cells are also important.

[0247] The term "anticancer therapy" refers to a therapy that can be used to treat cancer (e.g., lung cancer, such as NSCLC). Examples of anticancer therapeutic agents include, but are not limited to, immunomodulatory agents (e.g., agents that reduce or inhibit one or more immunosuppressive receptors (e.g., one or more immunosuppressive receptors selected from TIGIT, PD-L1, PD-1, CTLA-4, LAG3, TIM3, BTLA and / or VISTA), such as CTLA-4 antagonists, such as anti-CTLA-4 antagonist antibodies (e.g., ipilimumab). This includes, but is not limited to, anti-TIGIT antagonist antibodies or PD-1 axis binding antagonists (e.g., anti-PD-L1 antibodies), or agents that increase or activate one or more immune co-stimulatory receptors (e.g., one or more immune co-stimulatory receptors selected from CD226, OX-40, CD28, CD27, CD137, HVEM, and / or GITR, such as OX-40 agonists, e.g., OX-40 agonist antibodies), chemotherapeutic agents, growth inhibitors, cytotoxic agents, agents used in radiotherapy, anti-angiogenic agents, apoptosis agents, anti-microtubule agents, and other agents for treating cancer. Combinations of these are also included in this invention.

[0248] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents cell function and / or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., At). 211I 131 I 125 、Y 90 Re 186 Re 188 、Sm 153 Bi 212 P 32 Pb 212 Radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, doxorubicin, vinblastine alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitors; enzymes and fragments thereof such as lysozymes; antibiotics; toxins such as small molecule toxins or enzyme-active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof; and various antitumor or anticancer drugs disclosed below.

[0249] "Chemotherapy agents" include chemical compounds that can be used to treat cancer. Examples of chemotherapy agents include erlotinib (…). Genentech / OSIPharm.), Bortezomib Millennium Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), rhizobacterin, lactate dehydrogenase A (LDH-A), fulvestrant ( AstraZeneca), sunitib Pfizer / Sugen), letrozole ( Novartis, Imatinib Mesylate Novartis), finasunate Novartis, Oxaliplatin Sanofi), 5-FU (5-fluorouracil), leucovorin, rapamycin (Sirolimus), Wyeth, lapatinib GSK572016, Glaxo Smith Kline), Lonafamib (SCH 66336), Sorafenib ( Bayer Labs, Gefitinib ( AstraZeneca), AG1478, alkylating agents such as thiotepa and Cyclophosphamides; alkyl sulfonates, such as busulfan, indomethacin, and piperazine; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylmelamines, including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylomelamine; acetogen Inosine (especially bullatacin and bullatacinone); camptothecin (including topotecan and irinotecan); bryostatin; callystatin; CC-1065 (including its synthetic analogues adozelesin, carzelesin, and bizelesin); cryptophycins (especially nostocin 1 and nostocin 8); corticosteroids (including prednisone and prednisolone); cyproterone acetate; 5α-reductases (including finasteride and dutasteride); vorinostat, romidesin, ubiquistat, valproic acid, moxistatin, and dolastatin. dolastatin; interleukin, talc-based tyrosinase (including synthetic analogs KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards, such as chlorambucil, chlomaphazine, chlorphosphamide, estradiol, ifosfamide, methoxyfenozide, melphalan, neomycin, benzyl mustard cholesterol, prednimustine, trofosfamide, uracil mustard. Mustard; nitrosoureas, such as carmustine, chloramphenicol, timustine, lomustine, nimustine, and ranimnustine; antibiotics, such as enediyne antibiotics (such as calicheamicin, especially calicheamicin γ1I and calicheamicin ω1I (Angew Chem. Intl. Ed. Engl.)).1994 33:183-186); dynemicin, including dynemicin A; bisphosphonates, such as chlorophosphonates; esperamicin; and new dynemicin chromophores and related chromophore proteins (enediyne antibiotic chromophores), aclacnomysins, actinomycin, arbutycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, danomycin, detorubicin, 6-diazo-5-oxo-L-leucine. (Doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolidine-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin such as mitomycin C, mycophenolic acid, nogamycin, olivomycin, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozotocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues, such as folate (denofolate). Pterin, methotrexate, pteropterin, trimethotryxate; purine analogs, such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, fluxuridine; androgens, such as calcestrone, drotalbutone propionate, cyclothionol, meandranone, testrolide; antiadrenergics, such as aminoglutamine, mitotane, trilostane; folic acid supplements, such as folic acid; aceglatone; aldophosphamide glycosides. glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids, such as maytansine and ansamitocins;Mitoguazone; Mitoxantrone; Mopidamnol; Nitraerine; Pentostatin; Phenamet; Pirarubicin; Losoxantrone; Podophyllinic acid; 2-Ethylhydrazine; Methylbenzylhydrazine; Polysaccharide complexes (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2”-trichlorotriethylamine; trichothecene toxins (especially T-2 toxin, verracurin A, roridin A). A) and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactal; pipebroman; gacytosine; cytarabine (“Ara-C”); cyclophosphamide; thiotepa; taxanes, such as TAXOL (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ). (Excluding levofloxacin), albumin-engineered nanoparticle formulations of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.) and (Docetaxel, Sanofi-Aventis); Chloranmbucil; (Gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vincristine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; (Changchun Ruibin); Novantrone; Teniposide; Edatrexate; Daunomycin; Ampicillin; Capecitabine Ibandronate; CPT-11; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinols, such as retinoic acid; and any of the above-mentioned medicinal salts, acids and derivatives.

[0250] Chemotherapy agents also include: (i) anti-hormonal agents that modulate or inhibit the effects of hormones on tumors, such as anti-estrogens and selective estrogen receptor modulators (SERMs), including tamoxifen (including... Tamoxifen citrate, raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, raloxifene hydrochloride, LY117018, onapristone, and (ii) Inhibitors of aromatase, which regulates estrogen production in the adrenal glands, such as 4(5)-imidazole and aminoglutethimide. (medroxyprogesterone acetate) (Exemestane; Pfizer), Formestanie, Fadrozole) (vorozole) (letrozole; Novartis) and (iii) Antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, tripterelin, medroxyprogesterone acetate, diethylstilbestrol, Premarin, fluorometholone, all trans-retinoic acid, fenretinoin. (iv) protein kinase inhibitors (e.g., anaplastic lymphoma kinase (ALK) inhibitors, such as AF-802 (also known as CH-5424802 or alectinib)); (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, especially those that inhibit gene expression in signaling pathways associated with abnormal cell proliferation, such as PKC-α, Ralf, and H-Ras; (vii) ribozymes, such as VEGF expression inhibitors (e.g., trasatatabine (1,3-dioxane- ... (viii) HER2 expression inhibitors; (viii) vaccines, such as gene therapy vaccines, such as and rIL-2; topoisomerase 1 inhibitors, for example rmRH; and (ix) any of the above-mentioned medicinal salts, acids and derivatives.

[0251] Chemotherapy agents also include antibodies, such as alemtuzumab (Campath) and bevacizumab ( Genentech); Cetuximab ( Imclone; Panitumumab ( Amgen, rituximab Genentech / Biogen Idec, pertuzumab ( 2C4 (Genentech), trastuzumab ( Genentech, tositumomab (Bexxar, Corixia) and antibody drug conjugates, gemtuzumab ozomicin ( Wyeth). Other humanized monoclonal antibodies with therapeutic potential when used in combination with the compounds of the present invention as agents include: apolizumab, acelizumab, atlizumab, bapineuzumab, bivatuzumab mertansine, cantuzumab mertansine, cedelizumab, and pegylated certolizumab. pegol, cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin ozogamicin), ipilimumab, labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab, motovizumab, natazumab, nimotuzumab, nolovizumab, numavizumab, ocrelizumab, omalizumab, pallizumab, percoizumab, pecfusituzumab, pertuzumab, pexelizumab, ralivizumab, ranibizumab Reslivizumab, Reslizumab, Resyvizumab, Rovelizumab, Ruplizumab, Sibrotuzumab, Siplizumab, Sontuzumab, Tacatuzumab Tetraxetan, Tadocizumab, Talizumab, Tefibazumab, Tocilizumab, Toralizumab, TucotuzumabThe antibodies include celmoleukin, tucusituzumab, umavizumab, urtoxazumab, ustekinumab, visilizumab, and anti-interleukin-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), which is a genetically modified recombinant full-length IgG1λ antibody specifically designed for human sequences to recognize the interleukin-12p40 protein.

[0252] Chemotherapy agents also include “EGFR inhibitors,” which are compounds that bind to or directly interact with EGFR and inhibit or reduce its signaling activity, and are alternatively referred to as “EGFR antagonists.” Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see, U.S. Patent No. 4,943,533, Mendelsohn et al.) and their variants, such as chimeric 225 (C225 or cetuximab); ) and remodeled human 225 (H225) (see, WO 96 / 40210, Imclone Systems Inc.); IMC-11F8, a fully human antibody targeting EGFR (Imclone); antibodies binding type II mutant EGFR (US Patent No. 5,212,290); humanized and chimeric antibodies binding EGFR as described in US Patent No. 5,891,996; and human antibodies binding EGFR, such as ABX-EGF or panitumumab (see WO98 / 50433, Abgenix / Amgen); EMD 55900 (Stragliotto et al. Eur. J. Cancer) 32A:636-640 (1996)); EMD7200 (Maltozumab), a humanized EGFR antibody that competes with EGF and TGF-α for binding to EGFR (EMD / Merck); human EGFR antibody, HuMax-EGFR (GenMab); fully human antibodies, referred to as E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 and E7.6.3, and described in US 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 form immunoconjugates (see, for example, EP659,439A2, Merck Patent GmbH). EGFR antagonists include small molecules, such as those listed in U.S. Patent 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, Compounds described in WO98 / 14451, WO98 / 50038, WO99 / 09016, and WO99 / 24037, as well as 866,572, 6,399,602, 6,344,459, 6,344,455, 5,760,041, 6,002,008, and 5,747,498, and the following PCT publications: WO98 / 14451, WO98 / 50038, WO99 / 09016, and WO99 / 24037. Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib). Genentech / OSI Pharmaceuticals); PD183805 (CI 1033, 2-acrylamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib ( 4-(3'-chloro-4'-fluoroaniline)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca; ZM 105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluorophenyl)-N2-(1-methylpiperidin-4-yl)pyrimidinyl[5,4-d]pyrimidin-2,8-diamine, Boehringer Ingelheim) Ingelheim); PKI-166 ((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolidone[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidin); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide); EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolinyl]-4-(dimethylamino)-2-butenamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU 5271; Pfizer; dual EGFR / HER2 tyrosine kinase inhibitors, such as lapatinib ( GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2(methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine).

[0253] Chemotherapy agents also include "tyrosine kinase inhibitors," including the EGFR-targeted drugs mentioned above; insulin receptor tyrosine kinase inhibitors, including anaplastic lymphoma kinase (ALK) inhibitors, such as AF-802 (also known as CH-5424802 or alectinib), ASP3026, X396, LDK378, AP26113, and crizotinib. and ceritinib Small molecule HER2 tyrosine kinase inhibitors, such as TAK165 available from Takeda; CP-724,714, an orally selective inhibitor of ErbB2 receptor tyrosine kinase (Pfizer and OSI); dual HER inhibitors, such as EKB-569 (available from Wyeth), which preferentially binds to EGFR but simultaneously inhibits cells overexpressing HER2 and EGFR; lapatinib (GSK572016; available from GlaxoSmithKline), an orally administered HER2 and EGFR tyrosine kinase inhibitor; PKI-166 (available from Novartis); pan-HER inhibitors, such as canatinib (CI-1033; Pharmacia); Raf-1 inhibitors, such as the antisense agent ISIS-5132, which inhibits Raf-1 signaling, available from ISIS Pharmaceuticals; and non-HER-targeting TK inhibitors, such as imatinib mesylate (…). Available from GlaxoSmithKline); multi-target tyrosine kinase inhibitors, such as sunitinib ( Available from Pfizer); VEGF receptor tyrosine kinase inhibitors, such as vatalani (PTK787 / ZK222584, available from Novartis / Schering AG); MAPK extracellular kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines, such as PD 153035, 4-(3-chloroaniline)quinazoline; pyridopyrimidines; pyrimidines; pyrrolopyrimidines, such as CGP 59326, CGP 60261 and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin (difluoroformylmethane, 4,5-bis(4-fluoroaniline)phthalimide); tyrosine containing a nitrothiophene moiety; PD-0183805 (Warner-Lamber); antisense molecules (e.g., molecules that bind to HER-encoded nucleic acids); quinoxalines (US Patent No. 5,804,396); tyrosine phosphorylation inhibitors (US Patent No. 5,804,396); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schiring); pan-HER inhibitors, such as CI-1033 (Pfizer); Affinitac (ISIS 3521; ISIS / Lilly); imatinib mesylate PKI 166 (Novartis); GW2016 (GlaxoSmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); Sermatinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schiring); INC-1C11 (Imclone); Rapamycin (Sirolimus) ); or as described in any of the following patent publications: U.S. Patent Nos. 5,804,396, WO 1999 / 09016 (American Cyanamid), WO 1998 / 43960 (American Cyanamid), WO 1997 / 38983 (Warner Lambert), WO 1999 / 06378 (Warner Lambert), WO 1999 / 06396 (Warner Lambert), WO 1996 / 30347 (Pfizer, Inc.), WO 1996 / 33978 (Zeneca), WO 1996 / 3397 (Zeneca), and WO 1996 / 33980 (Zeneca).

[0254] Chemotherapy agents also include dexamethasone, interferon, colchicine, metoprine, cyclosporine, amphotericin B, metronidazole, alemtuzumab, alitretinoin, allopurinol, aifostine, arsenic trioxide, asparaginase, live BCG, bevacizumab, bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, and epioetin. Alfa, erlotinib, filgrastim, histrelinacetate, ibritumomab, interferon alpha-2a, interferon alpha-2b, lenalidomide, levamisole, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelvekin, palifermin, pamidronate, pegademase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium), quinacrine, rasburicase, sargramostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, ATRA, valrubicin, zoledronate, and zoledronic acid and their pharmaceutical salts.

[0255] Chemotherapy agents also include hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivalate, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, betamethasone, betamethasone sodium phosphate, dexamethasone, and dexamethasone sodium phosphate. Phosphate), fluocortolone, hydrocortisone-17-butyrate, hydrocortisone-17-valerate, aclometasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasol-17-propionate, fluocortolone caproate, fluocortolone pivalate, and fluprednidene acetate. acetate); immunoselective anti-inflammatory peptides (ImSAIDs), such as phenylalanine-glutamine-glycine (FEG) and its D-isomer (feG) (IMULANBioTherapeutics, LLC);Antirheumatic drugs, such as azathioprine, cyclosporine (cyclosporine A), D-penicillamine, gold salts, hydroxychloroquine, leflunomideminocycline, sulfasalazine; tumor necrosis factor α (TNFα) blockers, such as etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), cetuzumab (Cimzia), golimumab (Simponi); interleukin-1 (IL-1) blockers, such as kinetin; T-cell co-stimulatory blockers, such as abatacept (Orencia); interleukin-6 (IL-6) blockers, such as tocilizumab. Interleukin-13 (IL-13) blockers, such as lebrikizumab; interferon-alpha (IFN) blockers, such as rontalizumab; β7 integrin blockers, such as rhuMAbβ7; and IgE pathway blockers, such as Anti-M1. Prime; secreted homotrimer LTa3 and membrane-bound heterotrimer LTa1 / β2 blockers, such as antilymphotoxin α (LTa); radioisotopes (e.g., radioisotopes of At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212 and Lu); miscellaneous research reagents, such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH3 or farnesyltransferase inhibitors (L-739749, L-744832); polyphenols, such as quercetin, resveratrol, leucine, epigallocatechine gallate, theaflavins, flavanols, proanthocyanidins, betulinic acid and their derivatives; autophagy inhibitors, such as chloroquine; δ-9-tetrahydrocannabinol (dronabinol, ); β-lapachone; laparol; colchicine; betulinic acid; acetylcamptothecin, scopolectin, and 9-aminocamptothecin; podophyllotoxin; tegafur Besarodine Bisphosphonates, such as chlorophosphonates (such as...) or etidronate NE-58095, Zoledronic Acid / Zoledronate alendronate pamidronate tiludronate or risephosphonates and epidermal growth factor receptor (EGF-R); vaccines, for example Vaccines; perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), protein body inhibitors (e.g., PS341); CCI-779; Tipifarnib (R11577); orafenib, ABT510; Bcl-2 inhibitors, such as oblimersen sodium. Pixantrone; farnesyltransferase inhibitors, such as lonafarnib (SCH 6636, SARASARTM); and any of the pharmaceutical salts, acids, or derivatives thereof; and combinations of two or more of the above, such as CHOP, an abbreviation for combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone; and FOLFOX, oxaliplatin. TM (This is an abbreviation for a treatment regimen combining 5-FU and leucovorin.)

[0256] Chemotherapy agents also include nonsteroidal anti-inflammatory drugs (NSAIDs) with analgesic, antipyretic, and anti-inflammatory effects. NSAIDs include non-selective inhibitors of cyclooxygenase. Specific examples of NSAIDs include aspirin, propionic acid derivatives (e.g., ibuprofen, fenolofen, ketoprofen, flurbiprofen, oxaprozin, and naproxen), acetic acid derivatives (e.g., indomethacin, sulindac, etodoxacin, diclofenac), enolic acid derivatives (e.g., piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, and itoxicam), fenamic acid derivatives (e.g., mefenamic acid, meclofenamic acid, flufenamic acid, and tofenamic acid), and COX-2 inhibitors (e.g., celecoxib, etoricoxib, lumiracoxib, parecoxib, rofecoxib, rofecoxib, and valdecoxib). NSAIDs can be used to relieve symptoms of conditions such as rheumatoid arthritis, osteoarthritis, inflammatory arthritis, ankylosing spondylitis, psoriatic arthritis, Rett syndrome, acute gout, dysmenorrhea, migraine, headache and migraine, postoperative pain, mild to moderate pain due to inflammation and tissue damage, fever, intestinal obstruction and renal colic.

[0257] An “effective amount” of a compound, such as an anti-TIGIT antagonist antibody or a PD-1 axis binding antagonist (e.g., an anti-PD-L1 antibody) or a combination thereof (e.g., a pharmaceutical composition), is at least the minimum amount required to achieve a desired therapeutic outcome, such as a measurable increase in total survival or progression-free survival in a particular disease or condition (e.g., cancer, such as lung cancer (e.g., NSCLC)). The effective amount described herein can vary depending on factors such as the patient’s disease status, age, sex, and weight, and the antibody’s ability to elicit the expected response in the subject. An effective amount is also the amount in which the beneficial effect of treatment outweighs any toxic or adverse effects of treatment. For prophylactic use, beneficial or expected outcomes include elimination or reduction of risk, lessening of severity, or delay of disease onset, including biochemical, histological, and / or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes that occur during disease development. For therapeutic use, beneficial or expected outcomes include clinical outcomes such as reduction of one or more symptoms caused by the disease (e.g., cancer-related pain, reduction or delay of symptomatic skeletal-related events (SSEs), according to the European Organisation for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire). The reduction of symptoms of the QLQ-C30 (e.g., fatigue, nausea, vomiting, pain, shortness of breath, insomnia, loss of appetite, constipation, diarrhea, or general levels of physical, emotional, cognitive, or social functioning), pain relief as measured by, for example, a 10-point Numerical Rating Scale for Pain Severity (measured at its worst) (NRS), and / or a reduction of lung cancer-related symptoms as assessed by the Lung Cancer (SILC) Scale according to the Health Related Quality of Life (HRQoL) questionnaire (e.g., time to onset (TTD) of cough, shortness of breath, and chest pain), improves the quality of life of people with the disease. The effective amount of a drug may improve quality of life, reduce the dosage of other medications required to treat the disease, such as by targeting to enhance the effect of another drug, delay disease progression (e.g., progression-free survival or radiographic progression-free survival (rPFS)); delay definitive clinical progression (e.g., progression of cancer-related pain, symptomatic skeletal events, deterioration of Eastern Cooperative Group Oncology Group (ECOG) performance status (PS) (e.g., how the disease affects a patient's ability to perform daily activities), and / or the initiation of the next systemic anticancer therapy), and / or delay the time of lung-specific antigen progression) and / or prolong survival. In the case of cancer or tumor, an effective amount of the drug may reduce the number of cancer cells; reduce tumor size; inhibit (i.e., to some extent slow down or expect to stop) the infiltration of cancer cells into surrounding organs; inhibit (i.e., to some extent slow down and expect to stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with the condition to some extent. The effective amount may be administered once or multiple times. For the purposes of this invention, the effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to directly or indirectly prevent or treat the disease.As understood in a clinical context, an effective amount of a drug, compound, or drug composition may or may not be achieved when combined with another drug, compound, or drug composition. Therefore, an "effective amount" may be considered when administering one or more therapeutic agents, and an effective amount of a single agent may be considered if the desired result can be obtained or achieved when combined with one or more other agents.

[0258] "Individual response" or "response" may be assessed using any endpoint that indicates benefit to the subject, including but not limited to (1) inhibiting disease progression (e.g., progression of cancer such as lung cancer (e.g., NSCLC)) to some extent, including slowing and completely blocking it; (2) reducing tumor size; (3) inhibiting (i.e., reducing, slowing or completely stopping) cancer cell infiltration into adjacent surrounding organs and / or tissues; (4) inhibiting (i.e., reducing, slowing or completely stopping) metastasis; (5) alleviating one or more symptoms associated with the disease or ailment (e.g., cancer such as lung cancer (e.g., NSCLC)) to some extent; (6) increasing or prolonging survival, including overall survival and progression-free survival; and / or (9) reducing mortality at a given time point after treatment.

[0259] As used in this article, "complete remission" or "CR" refers to the disappearance of all target lesions.

[0260] As used in this article, "partial remission" or "PR" means a reduction of at least 30% in the sum of the longest diameters (SLDs) of the target lesions at baseline.

[0261] As used in this article, "Objective Response Rate" (ORR) refers to the sum of the complete response (CR) rate and the partial response (PR) rate.

[0262] A subject’s “effective response” to a drug or treatment, or a subject’s “responsiveness” and similar terms, refers to a clinical or therapeutically beneficial effect conferred on a subject who is at risk of or has a disease or condition such as cancer. In one embodiment, such benefit includes one or more of the following: prolonged survival (including overall survival and progression-free survival); objective remission (including complete remission or partial remission); or improvement of signs or symptoms of cancer.

[0263] Subjects who do not respond to treatment are those who do not have any of the following: prolonged survival (including overall survival and progression-free survival); objective response (including complete or partial response); or improvement in signs or symptoms of cancer.

[0264] The term "survival" refers to a patient still being alive, and includes overall survival and progression-free survival.

[0265] As used in this article, “overall survival” (OS) refers to the percentage of subjects who survive a specific duration, such as 1 year or 5 years from the time of diagnosis or treatment.

[0266] As used in this article, “progression-free survival (PFS)” refers to the length of time during and after treatment when the treated disease (e.g., cancer, such as lung cancer (e.g., NSCLC)) does not worsen. PFS can include the amount of time a patient experiences a complete or partial remission, as well as the amount of time a patient experiences stable disease.

[0267] As used in this article, "stable disease" or "SD" refers to the minimum SLD since the start of treatment, where the target lesion has neither shrunk sufficiently to meet PR nor increased sufficiently to meet PD.

[0268] As used in this article, “progressive disease” or “PD” means an increase of at least 20% in the target lesion’s SLD, with reference to the minimum SLD recorded since the start of treatment or the appearance of one or more new lesions.

[0269] As used herein, “delayed progression” of a disease or ailment means the slowing, hindering, mitigating, delaying, stabilizing, and / or postponing of the development of a disease or ailment (e.g., cancer, such as lung cancer (e.g., NSCLC)). Such delay can vary in length depending on medical history and / or the subject to be treated. It will be apparent to those skilled in the art that adequate or significant delay can effectively encompass prevention, as the subject will not develop the disease. For example, in advanced cancer, the development of central nervous system (CNS) metastases may be delayed.

[0270] "Prolonged survival" refers to an increase in overall survival or progression-free survival in treated patients relative to untreated patients (e.g., patients not receiving drug treatment), or patients not expressing a biomarker at a specified level, and / or patients treated with an approved antitumor agent. Objective response refers to a measurable response, including complete response (CR) or partial response (PR).

[0271] As used herein, “hazard ratio” or “HR” is a statistical definition of the incidence of an event. For the purposes of this invention, the hazard ratio is defined as the probability of an event (e.g., PFS or OS) in the experimental (e.g., treatment) group / arm divided by the probability of the event in the control group / arm. An HR of 1 indicates that the relative risk of the endpoint (e.g., death) is equal in the treatment and control groups; a value greater than 1 indicates that the risk is greater in the treatment group than in the control group; a value less than 1 indicates that the risk is greater in the control group than in the treatment group. The “hazard ratio” in a progression-free survival analysis (i.e., PFS HR) summarizes the difference between two progression-free survival curves, representing the reduced risk of death with treatment compared to the control during the follow-up period. The “hazard ratio” in an overall survival analysis (i.e., OS HR) summarizes the difference between two overall survival curves, representing the reduced risk of death with treatment compared to the control during the follow-up period.

[0272] As used herein, the “Ventana SP263 IHC assay” (referred to herein as the Ventana SP263 CDx assay) was performed according to the Ventana PD-L1 (SP263) assay package insert (Tucson, AZ: Ventana Medical Systems, Inc.), which is incorporated herein by reference in its entirety.

[0273] As used herein, the “Ventana SP142 IHC assay” was performed according to the Ventana PD-L1 (SP142) assay package insert (Tucson, AZ: Ventana Medical Systems, Inc.), which is incorporated herein by reference in its entirety.

[0274] As used herein, the “pharmDx 22C3 IHC assay” was performed according to the PD-L1 IHC 22C3pharmDx package insert (Carpinteria, CA: Dako, Agilent Pathology Solutions), which is incorporated herein by reference in its entirety.

[0275] As used herein, “tumor-infiltrating immune cells” refers to any immune cell present in a tumor or its sample. Tumor-infiltrating immune cells include, but are not limited to, intratumoral immune cells, peritumoral immune cells, other tumor stromal cells (e.g., fibroblasts), or any combination thereof. Such tumor-infiltrating immune cells can be, for example, T lymphocytes (such as CD8+ T lymphocytes and / or CD4+ T lymphocytes), B lymphocytes or other bone marrow lineage cells, including granulocytes (e.g., neutrophils, eosinophils, and basophils), monocytes, macrophages, dendritic cells (e.g., finger dendritic cells), histiocytes, and natural killer cells.

[0276] As used herein, the term "biomarker" refers to an indicator that can be detected in a sample, such as predictive, diagnostic, and / or prognostic indicators. In some embodiments, the biomarker is a gene. Biomarkers include, but are not limited to, peptides, polynucleotides (e.g., DNA and / or RNA), polynucleotide copy number alterations (e.g., DNA copy number), peptide and polynucleotide modifications (e.g., post-translational modifications), carbohydrates, and / or glycolipid-based molecular markers.

[0277] The term "antibody" includes monoclonal antibodies (including full-length antibodies having the Fc region of an immunoglobulin), antibody compositions with multi-epitope specificity, multispecific antibodies (e.g., bispecific antibodies), bispecific antibodies, and single-chain molecules, as well as antibody fragments, including antigen-binding fragments such as Fab, F(ab')2, and Fv. The term "immunoglobulin" (Ig) is used interchangeably with "antibody" herein.

[0278] The basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. IgM antibodies consist of five basic heterotetrameric units plus an additional polypeptide called the J chain and contain 10 antigen-binding sites, while IgA antibodies contain 2–5 basic 4-chain units that can combine with the J chain to polymerize into multivalent conjugates. In the case of IgG, the 4-chain unit is typically about 150,000 Daltons. Each L chain is linked to the H chain by a covalent disulfide bond, and two H chains are linked to each other by one or more disulfide bonds depending on the homotype of the H chains. Each H and L chain also has regularly spaced intrachain disulfide bonds. Each H chain has a variable domain (V) at its N-terminus. H ), followed by three constant structure domains (C H (For each α chain and γ chain) and four C H Structural domains (for μ and ε types). Each L chain has a variable structural domain (V) at the N-terminus. L ), and has a constant structural domain at the other end. V L With V H Align, and CL With the first constant structural domain (C) of the heavy chain H 1) Alignment. Specific amino acid residues are believed to form interfaces between the variable domains of the light and heavy chains. V H and V L The pairs together form a single antigen-binding site. For information on the structure and properties of different classes of antibodies, see, for example, Basic and Clinical Immunology, 8th ed., Daniel P. Stites, Abba I. Terr, and Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994, p. 71 and Chapter 6. L-chains derived from any vertebrate, based on the amino acid sequence of their constant domain, can be classified into one of two distinct types, called kappa (κ) and lamuda (λ). Immunoglobulins, based on the amino acid sequence of their heavy chain constant domain (CH), can be classified into different classes or isotypes. Five classes of immunoglobulins exist: IgA, IgD, IgE, IgG, and IgM, each possessing a heavy chain called α, δ, ε, γ, and μ, respectively. The γ and α classes are further subdivided into subclasses based on relatively small differences in CH sequence and function. For example, humans express the following subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2.

[0279] As used herein, the term "hypervariant region" or "HVR" refers to the regions within the variable domain of an antibody that are hypervariable in sequence and determine antigen-binding specificity, such as the "complementarity-determining region" ("CDR").

[0280] Typically, an antibody contains six CDRs; three in the VH (CDR-H1, CDR-H2, CDR-H3) and three in the VL (CDR-L1, CDR-L2, CDR-L3). Exemplary CDRs in this document include:

[0281] (a) Highly variable rings appearing at the following amino acid residues: 26 to 32 (L1), 50 to 52 (L2), 91 to 96 (L3), 26 to 32 (H1), 53 to 55 (H2) and 96 to 101 (H3) (Chothia and Lesk, J. Mol. Biol. 196: 901-917 (1987));

[0282] (b) CDRs appearing at the following amino acid residues: 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991)); and

[0283] (c) Antigen contact sites appearing at the following amino acid residues: 27c to 36 (L1), 46 to 55 (L2), 89 to 96 (L3), 30 to 35b (H1), 47 to 58 (H2), and 93 to 101 (H3) (MacCallum et al., J.Mol.Biol.262:732-745 (1996)).

[0284] Unless otherwise specified, the CDR is determined according to the method described by Kabat et al., citing above. Those skilled in the art will understand that the CDR name may also be determined according to the method described by Chothia, citing above, the method described by McCallum, citing above, or any other scientifically accepted naming system.

[0285] The expressions “Kabat’s variable domain residue number” or “Kabat’s amino acid position number” and their variations refer to the numbering system for heavy chain or light chain variable domains used in antibody compilation in the aforementioned Kabat et al. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids, corresponding to shortening or insertion of the FR or HVR of the variable domain. For example, the heavy chain variable domain may include a single amino acid insertion after residue 52 of H2 (residue 52a according to Kabat numbering) and inserted residues after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c, etc. according to Kabat numbering). The Kabat number of residues for a given antibody can be determined by comparing the antibody sequence with homologous regions of a “standard” Kabat-numbered sequence.

[0286] The term "variable" refers to the significant differences in certain segments of the variable domain across antibody sequences. The V domain mediates antigen binding and defines the specificity of a particular antibody for its specific antigen. However, variability is not uniformly distributed across the entire span of the variable domain. Instead, it is concentrated in three segments called hypervariable regions (HVRs) within the variable domains of both the light and heavy chains. The more conserved portions of the variable domain are called frame regions (FRs). The variable domains of the natural heavy and light chains each contain four FRs, which primarily employ a β-sheet structure linked by three HVRs that form loops connecting the β-sheet structure and, in some cases, form part of the β-sheet structure. The HVRs in each chain are tightly held together by the FRs and, together with the HVRs in the other chain, contribute to the formation of the antibody's antigen-binding site (see Kabat et al., Sequences of Immunological Interest, 5th ed., National Institute of Health, Bethesda, MD (1991)). Constant domains do not directly participate in antibody-antigen binding but exhibit various effector functions, such as antibody involvement in antibody-dependent cytotoxicity.

[0287] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of either the heavy or light chain. The variable domains of the heavy and light chains are referred to as "VH" and "VL," respectively. These domains are typically the most variable parts of an antibody (relative to other antibodies in the same class) and contain the antigen-binding site.

[0288] "Frame" or "FR" refers to the variable domain residues other than the hypervariable region (HVR) residues. A variable domain FR typically consists of four FR domains: FR1, FR2, FR3, and FR4. Therefore, the HVR and FR sequences usually appear in the VH (or VL) as follows: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0289] The terms "full-length antibody," "intact antibody," and "whole antibody" are used interchangeably and refer to an antibody in its essentially complete form rather than an antibody fragment. Specifically, intact antibodies include those having both a heavy chain and a light chain that includes an Fc region. The constant domain can be a native sequence constant domain (e.g., a human native sequence constant domain) or a variant of its amino acid sequence. In some cases, intact antibodies may have one or more effector functions.

[0290] An "antibody fragment" comprises a portion of a complete antibody, preferably comprising the antigen-binding and / or variable regions of the complete antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; bisomatic antibodies; and linear antibodies (see U.S. Patent 5,641,870, Example 2; Zapata et al., Protein Eng.). 8(10) :1057-1062

[1995] ); single-chain antibody molecules and multispecific antibodies formed from antibody fragments. Papain digests the antibody to produce two identical antigen-binding fragments (called “Fab” fragments) and a residual “Fc” fragment (the name reflects its ability to crystallize easily). The Fab fragment consists of the intact L chain and the variable region domain (V) of the H chain. H ) and the first constant structural domain of a heavy chain (C H 1) Composition. Each Fab fragment is monovalent in terms of antigen binding, meaning it has a single antigen-binding site. Pepsin treatment of the antibody produces a single large F(ab')2 fragment, which roughly corresponds to two Fab fragments linked by disulfide bonds that have different antigen-binding activities and are still capable of cross-linking the antigen. The difference between the Fab' fragment and the Fab fragment lies in the fact that the Fab' fragment has a C... H Additional residues were added to the carboxyl terminus of the 1 domain, including one or more cysteine ​​residues from the antibody hinge region. Fab'-SH is the designation used herein for Fab' in which the cysteine ​​residues of 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 hinge cysteine ​​residue in between. Other chemical conjugations of antibody fragments are also known.

[0291] The Fc fragment comprises the carboxyl-terminal portions of two H chains linked together by disulfide bonds. The effector function of an antibody is determined by a sequence in the Fc region, which is also recognized by Fc receptors (FcRs) present on certain types of cells.

[0292] The "functional fragment" of the antibody of this invention comprises a portion of the complete antibody, typically including the antigen-binding region or variable region of the complete antibody, or the Fc region of an antibody that retains or has modified FcR binding ability. Examples of antibody fragments include linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0293] "Fv" is the smallest antibody fragment containing both a complete antigen recognition and binding site. This fragment consists of a tightly bound, non-covalently linked dimer of a heavy chain variable region domain and a light chain variable region domain. Folding of these two domains produces six hypervariable rings (three rings from the H chain and three from the L chain), which contribute amino acid residues to achieve antigen binding, thus giving the antibody antigen-binding specificity. However, even a single variable domain (or half of an Fv containing only three antigen-specific HVRs) can recognize and bind antigens, although with lower affinity than the complete binding site.

[0294] "Single-chain Fv", also abbreviated as "sFv" or "scFv", is a type of Fv containing a single polypeptide chain linked to a V-chain. H and V L Antibody fragments containing antibody domains. Preferably, the sFv polypeptide is in V H and V L The domains are further joined by polypeptide linkers, enabling sFv to form the desired antigen-binding structure. For a review of sFv, see Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, edited by Rosenburg and Moore, Springer-Verlag, New York, pp. 269-315, 1994.

[0295] The term "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain, which includes both native sequence Fc regions and variant Fc regions. Although the boundaries of the immunoglobulin heavy chain Fc region may vary, the human IgG heavy chain Fc region is generally defined as extending from an amino acid residue at position Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) may be removed, for example, during antibody production or purification or by recombinantly designing the nucleic acid encoding the antibody heavy chain. Therefore, compositions of complete antibodies may include antibody populations with all K447 residues removed, antibody populations with K447 residues not removed, and antibody populations having mixtures containing and without K447 residues. Suitable native sequence Fc regions for the antibodies of this invention include human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4. Unless otherwise stated herein, amino acid residues in the Fc region or constant region are numbered according to the EU numbering system (also known as the EU index), as described by Kabat et al. (Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991) (see above).

[0296] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. Preferred FcRs are native human FcRs. Furthermore, preferred FcRs are receptors that bind to IgG antibodies (γ receptors) and include subclasses of FcγRI, FcγRII, and FcγRIII, including allelic variants and alternative spliced ​​forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences, differing primarily in their cytoplasmic domains. The activating receptor FcγRIIA contains an activation motif (ITAM) based on the immunoreceptor tyrosine residue in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an inhibitory motif (ITIM) based on the immunoreceptor tyrosine residue in its cytoplasmic domain. (See M.) Annu.Rev.Immunol. 15 :203-234 (1997). For a review of FcR, see: Ravetch and Kinet, Annu. Rev. Immunol. 9 :457-92(1991); Capel et al., Immunomethods 4 :25-34 (1994); and deHaas et al., J.Lab.Clin.Med.126 :330-41 (1995). The term “FcR” in this paper covers other FcRs, including those to be identified in the future.

[0297] The term bisomatic antibody refers to a small antibody fragment prepared by constructing an sFv fragment (see previous paragraph), in which V... H and V L The domains are connected by short linkers (approximately 5-10 residues), enabling interchain rather than intrachain pairing of the V domains, resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites. Bispecific bimeric antibodies are heterodimers of two "crossed" sFv fragments, where the V domains of the two antibodies... H and V L The domains are located on different polypeptide chains. Biantibodies are described in more detail, for example, EP 404,097; WO 93 / 11161; Hollinger et al., Proc. Natl. Acad. Sci. USA 90 :6444-6448(1993).

[0298] The monoclonal antibodies described herein specifically include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy and / or light chains is identical or homologous to the corresponding sequences in antibodies from a specific species or belonging to a specific antibody class or subclass, while the remainder of one or more chains is identical or homologous to the corresponding sequences in antibodies from another species or belonging to another antibody class or subclass, as well as fragments of those antibodies, provided they exhibit the desired biological activity (US Patent No. 4,816,567; Morrison et al., Proc. Natl. Acad. Sci. USA). 81 :6851-6855(1984)). The purpose of this article is to include chimeric antibodies including Antibodies, wherein the antigen-binding region of an antibody is derived from an antibody produced, for example, by immunizing a macaque with a target antigen. As used herein, “humanized antibody” is used as a subset of “chimeric antibody”.

[0299] An antibody's "class" refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some of these antibodies can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The constant domains of the heavy chain corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0300] “Affinity” refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen, such as TIGIT or PD-L1). Unless otherwise stated, as used herein, “binding affinity” refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its partner Y can generally be determined by the dissociation constant (K). D ( ) indicates. Affinity can be measured by conventional methods known in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0301] "Human antibody" is an antibody having an amino acid sequence corresponding to that of an antibody produced by a human and / or prepared using any of the techniques disclosed herein for preparing human antibodies. This definition of human antibody specifically excludes humanized antibodies containing non-human antigen-binding residues. Human antibodies can be produced 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). Methods also available for preparing human monoclonal antibodies include those described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5 :368-74 (2001). Human antibodies can be prepared by administering an antigen to a transgenic animal that has been modified to produce such antibodies in response to antigen challenge, but whose endogenous loci have been deactivated, for example, in immunized xenogeneic mice (see, for example, on XENOMOUSE). TM The technology is referenced in U.S. patents 6,075,181 and 6,150,584. See also, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) concerning human antibodies produced via human B-cell hybridoma technology.

[0302] "Humanized" forms of non-human (e.g., mouse) antibodies are chimeric antibodies containing minimal sequences derived from non-human immunoglobulins. In one embodiment, a humanized antibody is a human immunoglobulin (receptor antibody) in which residues from the receptor HVR (as defined below) are replaced by residues from the HVR of a non-human species (donor antibody) such as mouse, rat, rabbit, or a non-human primate with the desired specificity, affinity, and / or ability. In some cases, framework ("FR") residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may contain residues not present in the receptor or donor antibody. These modifications can be made to further improve antibody performance, such as binding affinity. Generally, a humanized antibody will contain at least one, typically two variable domains, wherein all or substantially all of the hypervariable loops correspond to those of the non-human immunoglobulin sequence, and all or substantially all of the FR regions are those of the human immunoglobulin sequence, although the FR regions may include one or more individual FR residue substitutions to improve antibody performance, such as binding affinity, isomerization, immunogenicity, etc. The number of these amino acid substitutions in the FR is typically no more than 6 in the H chain and no more than 3 in the L chain. Humanized antibodies will also optionally contain at least a portion of the immunoglobulin constant region (Fc), which is typically a 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). See also, for example, 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.

[0303] When used to describe the various antibodies disclosed herein, the term "isolated antibody" refers to an antibody that has been identified and isolated and / or recovered from the cells or cell cultures in which it is expressed. Its natural environmental contaminant components are materials that typically interfere with the diagnostic or therapeutic use of the peptide and may include enzymes, hormones, and other protein or non-protein solutes. In some embodiments, the antibody is purified to a purity greater than 95% or 99%, as determined by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed-phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87 (2007). In preferred embodiments, the antibody will be purified to (1) a degree sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence using a rotary cup sequencer, or to (2) homogenized by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue or, preferably, silver staining. The isolated antibodies include recombinant intracellular in situ antibodies because at least one component of the peptide's native environment is absent. However, typically, the isolated peptide will be prepared through at least one purification step.

[0304] As used herein, the term "monoclonal antibody" refers to an antibody derived from a substantially homogeneous group of antibodies; for example, the individual antibodies contained in this group are identical, except for a small number of naturally occurring mutations and / or post-translational modifications (e.g., heteropolymerization, amidation). Monoclonal antibodies exhibit high specificity for a single antigenic site. In contrast to polyclonal antibody formulations, which typically comprise different antibodies targeting different determinants (epitopes), each monoclonal antibody targets a single determinant on the antigen. In addition to their specificity, monoclonal antibodies have the advantage of being synthesized via hybridoma culture without contamination from other immunoglobulins. The modifier "monoclonal" indicates that the antibody is derived from a substantially homogeneous group of antibodies and should not be interpreted as requiring the production of the antibody through any particular method. For example, the monoclonal antibodies used according to the present invention can be prepared by a variety of techniques, including, 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 edition, 1988); Hammerling et al., Monoclonal Antibodies and T-Cell Hybridomas). 563-681 (Elsevier, NY, 1981), recombinant DNA methods (see, for example, US Patent No. 4,816,567), phage display techniques (see, for example, Clackson et al., Nature, 352:624-628 (1991); Marks et al., J. Mol. Biol. 222:581-597 (1992); Sidhu et al., J. Mol. Biol. 338(2):299-310 (2004); Lee et al., J. Mol. Biol. 340(5):1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34):12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2):119-132(2004)) and techniques for producing human antibodies or human-like antibodies in animals having part or all of a human immunoglobulin locus or gene encoding a human immunoglobulin sequence (see, for example, WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 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); US 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 and Huszar, Intern. Rev. Immunol. 13:65-93 (1995).

[0305] The "percentage of amino acid sequence identity (%)" relative to a reference polypeptide sequence is defined as the percentage of amino acid residues in the candidate sequence that are identical to those in the reference polypeptide sequence after aligning the candidate sequence with the reference polypeptide sequence and introducing vacancies (if necessary) to achieve the maximum percentage of sequence identity, without considering any conserved substitutions as part of the sequence identity. Alignment used to determine the percentage of amino acid sequence identity can be performed in various ways within the scope of the art, such as using publicly available computer software, such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm required to achieve maximum alignment across the full length of the sequences being compared. However, for the purposes of this document, the sequence comparison computer program ALIGN-2 is used to generate the amino acid sequence identity value %. The ALIGN-2 sequence comparison computer program was written by Genentech, Inc., and the source code has been submitted with user documentation to the US Copyright Office, Washington DC, 20559, where it is registered under US Copyright Registry 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 should be compiled for use on UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and remain unchanged.

[0306] When using ALIGN-2 for amino acid sequence comparison, the percentage of identity between a given amino acid sequence A and or with respect to a given amino acid sequence B (which can be alternatively expressed as a given amino acid sequence A having or containing a percentage of identity with or with respect to a given amino acid sequence B) is calculated as follows:

[0307] 100 multiplied by the fraction X / Y

[0308] Where X is the number of amino acid residues that are scored as identical matches by the sequence alignment program ALIGN-2 in the alignment of A and B, and Y is the total number of amino acid residues in B. It should 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 % between A and B will not be equal to the amino acid sequence identity % between B and A. Unless otherwise specifically stated, all amino acid sequence identity % values ​​used herein were obtained using the ALIGN-2 computer program as described in the preceding paragraph.

[0309] As used herein, "subject" or "individual" means mammal, including but not limited to human or non-human mammals such as cattle, horses, dogs, sheep, or cats. In some implementations, the subject is a person. Patients are also subjects in this document.

[0310] As used herein, the term "sample" means a composition obtained or derived from a subject and / or individual intended for use, which contains, for example, cells and / or other molecular entities to be characterized and / or identified based on physical, biochemical, chemical, and / or physiological characteristics. For example, the phrases "tumor sample," "disease sample," and variations thereof refer to any sample obtained from a subject intended for use that is expected or known to contain cells and / or molecular entities to be characterized. In some embodiments, the sample is a tumor tissue sample (e.g., a lung cancer sample (e.g., an NSCLC 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, breast milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, feces, tumor lysis products, and tissue culture media, tissue extracts such as homogenized tissue, tumor tissue, cell extracts, and combinations thereof.

[0311] The terms “tissue sample” and “cell sample” refer to a collection of similar cells obtained from the tissue of a subject or individual. The source of a tissue or cell sample can be solid tissue from fresh, frozen, and / or preserved organs, tissue samples, biopsies, and / or aspirates; blood or any blood component, such as plasma; body fluids, such as cerebrospinal fluid, amniotic fluid, peritoneal fluid, or interstitial fluid; or cells from any time during the subject’s pregnancy or development. Tissue samples can also be primary or cultured cells or cell lines. Optionally, tissue or cell samples are obtained from diseased tissue / organ. Tissue samples may contain compounds that are naturally occurring and do not mix with tissues, such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, etc.

[0312] 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 disease-free portion (e.g., tissue or cell) of the same subject’s body. For example, healthy and / or disease-free cells or tissue are 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 body tissue and / or cells of the same 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 disease-free portion (e.g., tissue or cell) of a subject who is not that subject. In still 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 an individual body part that is not that subject.

[0313] Unless otherwise specified, the term "protein" as used herein means any naturally occurring protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term includes "full-length" unprocessed proteins, as well as any form of protein produced through cellular processing. The term also covers naturally occurring protein variants, such as splice variants or allele variants.

[0314] The terms “polynucleotide” or “nucleic acid” used interchangeably herein refer to a polymer of nucleotides of any length, including DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogues, or any substrate that can be incorporated into the polymer by DNA or RNA polymerase or by a synthetic reaction. Thus, for example, 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 DNA and RNA (which may be single-stranded, or more commonly double-stranded, or containing single-stranded and double-stranded regions). Additionally, the term “polynucleotide” as used herein refers to a triple-stranded region containing RNA or DNA, or both RNA and DNA. The strands in such regions may originate from the same molecule or from different molecules. A region may contain all of one or more of a molecule, but typically only a region of a portion of a molecule. One of the molecules having a triple-helical region is typically an oligonucleotide. The terms “polynucleotide” and “nucleic acid” specifically include mRNA and cDNA.

[0315] Polynucleotides can contain modified nucleotides, such as methylated nucleotides and their analogues. If present, the nucleotide structure can be modified before or after polymer assembly. The sequence of the nucleotide can be interrupted by non-nucleotide components. Polynucleotides can be further modified post-synthesized, for example, by conjugation with a tag. Other types of modifications include, for example, "capping," substitution of one or more naturally occurring nucleotides with analogues, internucleotide modifications such as those with non-electrically linked groups (e.g., methyl phosphate, triphosphate, phosphoamidate, carbamate, etc.) and those with electrically linked groups (e.g., thiophosphate, dithiophosphate, etc.), those containing overhanging moieties such as proteins (e.g., nucleases, toxins, antibodies, signal peptides, poly-L-lysine, etc.), those with intercalating agents (e.g., acridine, psoralen, etc.), those containing chelating agents (e.g., metals, radioactive metals, boron, oxidizing metals, etc.), those containing alkylating agents, those with modified links (e.g., α-terminal isomeric nucleic acids), and unmodified forms of polynucleotides. Furthermore, any hydroxyl groups typically present in the sugar can be substituted (e.g., phosphate ester groups, phosphate groups), protected by standard protecting groups, or activated to prepare additional linkages with other nucleotides, or can be conjugated to a solid or semi-solid support. The OH groups at the 5' and 3' ends can be phosphorylated or partially substituted by amine or organic end-capping groups of 1 to 20 carbon atoms. Other hydroxyl groups can also be derived as standard protecting groups. The polynucleotide can also contain similar forms of ribose or deoxyribose known in the art, including, for example, 2'-O-methyl-, 2'-O-allyl-, 2'-fluoro-, or 2'-azidoribose; carbocyclic sugar analogs; α-terminal isomers; epimeric sugars such as arabinose, xylose, or lythose; pyranoses; furanoses; sedoheptulose; acyclic analogs; and non-base nucleoside analogs such as methylribonucleotides. One or more phosphodiester bonds can be replaced by alternative linking groups. These alternative linking groups include, but are not limited to, aspects in which the phosphate is replaced by P(O)S (“thioester”), P(S)S (“dithioester”), (O)NR2 (“amic acid ester”), P(O)R, P(O)OR', CO, or CH2 (“methylal”), wherein each R or R' is independently H or a substituted or unsubstituted alkyl group (1-20C), optionally comprising an ether (-O-) linker, aryl, alkenyl, cycloalkyl, cycloalkenyl, or araldyl. Not all links in a polynucleotide need to be identical. The preceding description applies to all polynucleotides referred to herein, including RNA and DNA.

[0316] As used herein, “carrier” includes pharmaceutical carriers, excipients, or stabilizers that are non-toxic to cells or mammals exposed to them at the doses and concentrations employed. Physiologically acceptable carriers are typically pH-buffered aqueous solutions. Examples of physiologically acceptable carriers include: buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) peptides; 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. TM Polyethylene glycol (PEG) and PLURONICS TM .

[0317] The phrase “medicinal” means that the substance or composition must be chemically and / or toxicologically compatible with other ingredients in the preparation and / or with the mammals used for treatment.

[0318] The term "pharmaceutical formulation" refers to a formulation in which the bioactive ingredient contained therein is in a form in which the activity is effective and which does not contain any additional components that would have unacceptable toxicity to a subject to whom the formulation will be administered.

[0319] III. Prognostic methods and assays

[0320] A. Tumor-associated macrophage (TAM) genes and gene characteristics

[0321] Methods for identifying individuals who can benefit from treatment

[0322] (i) TAM gene

[0323] In one aspect, the present invention provides a method for identifying an individual with cancer (e.g., lung cancer, e.g., non-small cell lung cancer (NSCLC)) who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tireliumab)), the method comprising detecting one or more of the following tumor-associated macrophages (TAMs) (e.g., one, two, three, four of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO) in a sample from the individual. The expression levels of the following six genes (complement C1q subunit C (C1QC), macrophage scavenger receptor type I and II (MSR1), macrophage mannose receptor 1 (MRC1), V-set and immunoglobulin domain-containing protein 4 (VSIG4), secreted phosphoprotein 1 (SPP1), and collagen-structured macrophage receptor (MARCO) were used to identify individuals who could benefit from treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies.

[0324] (ii) TAM characteristic scores

[0325] On the other hand, the present invention provides a method for identifying an individual with cancer (e.g., lung cancer, such as NSCLC) who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tirelinumab)), the method comprising detecting the expression levels of at least two of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO (e.g., two, three, four, five, or all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO) in a sample from the individual and determining a tumor-associated macrophage (TAM) eigenvalue score therefrom, wherein a TAM eigenvalue score higher than a reference TAM eigenvalue score identifies the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist.

[0326] On the other hand, the present invention provides a method for identifying an individual with cancer (e.g., lung cancer, such as NSCLC) who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tirelinumab)), the method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from the individual and determining a TAM characteristic score therefrom, wherein a TAM characteristic score higher than a reference TAM characteristic score identifies the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0327] In some aspects, the individual has a TAM characteristic score higher than a reference TAM characteristic score in the sample, and the method further includes administering an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody to the individual. Exemplary methods for determining TAM characteristic scores and exemplary reference TAM characteristic scores are provided below. Exemplary PD-1 axis-binding antagonists, anti-TIGIT antagonist antibodies, and treatment methods comprising these agents are provided in Section IV.

[0328] In some respects of any of the methods presented in this article, the cancer is lung cancer, such as NSCLC, small cell lung cancer (SCLC), or lung carcinoid tumor. In some respects, the individual is a person.

[0329] Methods for selecting a therapy

[0330] (i) TAM gene

[0331] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising detecting the expression levels of one or more of the tumor-associated macrophage (TAM) genes C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO (e.g., one, two, three, four, five, or all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO) in a sample from the individual, wherein the expression levels of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO above their respective reference expression levels identify the individual as an individual who may benefit from treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0332] (ii) TAM characteristic scores

[0333] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising detecting the expression levels of at least two of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO (e.g., two, three, four, five, or all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO) in a sample from the individual and determining a TAM characteristic score therefrom, wherein a TAM characteristic score higher than a reference TAM characteristic score identifies the individual as an individual who may benefit from treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0334] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from the individual and determining a TAM characteristic score therefrom, wherein a TAM characteristic score higher than a reference TAM characteristic score identifies the individual as an individual who may benefit from treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0335] In some respects, the individual has a TAM characteristic score higher than the reference TAM characteristic score in the sample, and the method further includes administering an effective amount of PD-1 axis-binding antagonist and anti-TIGIT antagonist antibody to the individual.

[0336] Treatment

[0337] (i) TAM gene

[0338] On the other hand, the present invention provides a method for treating an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising (a) detecting the expression levels of one or more of the tumor-associated macrophage (TAM) genes C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO (e.g., one, two, three, four, five, or all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO) in a sample from the individual, wherein C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO are expressed in a sample from the individual. 1. The expression level of one or more of VSIG4, SPP1 and MARCO is higher than their respective reference expression levels, thereby identifying an individual as an individual who may benefit from treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies; and (b) administering to the individual an effective amount of PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies (e.g., PD-1 axis binding antagonists disclosed in Section IV of this document (e.g., atezolizumab) and anti-TIGIT antagonist antibodies disclosed in Section IV of this document (e.g., tislelizumab)).

[0339] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tisreliumab)), wherein the individual has been determined to have expression levels of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO above their respective reference expression levels, thereby identifying the individual as an individual who may benefit from treatment including the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

[0340] (ii) TAM characteristic scores

[0341] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising (a) detecting the expression levels of at least two of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO (e.g., two, three, four, five, or all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO) in a sample from the individual and determining a TAM characteristic score therefrom, wherein the TAM characteristic score is higher than a reference TAM characteristic score, and thereby identifying the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody; and (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0342] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising (a) detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from the individual and determining a TAM characteristic score therefrom, wherein the TAM characteristic score is higher than a reference TAM characteristic score, and thereby identifying the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody; and (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0343] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising administering to the individual a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tisreliumab)), wherein the individual has been determined to have a TAM characteristic score higher than a reference TAM characteristic score, thereby identifying the individual as an individual who may benefit from treatment including the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody, and wherein the TAM characteristic score is based on the expression level of at least two of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO (e.g., two, three, four, five or all six of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO) detected in a sample from the individual.

[0344] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tisreliumab)), wherein the individual has been determined to have a TAM characteristic score higher than a reference TAM characteristic score, thereby identifying the individual as an individual who may benefit from treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, and wherein the TAM characteristic score is based on the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO detected in a sample from the individual.

[0345] Benefit

[0346] Individuals who benefit from treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies may experience, for example: delaying or preventing the onset or recurrence of cancer (e.g., lung cancer, such as NSCLC), alleviating cancer symptoms, reducing any direct or indirect pathological consequences of cancer, preventing metastasis, slowing the rate of disease progression, improving or alleviating the condition, and alleviating or improving prognosis.

[0347] In some respects, the benefits achieved through treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies are increased progression-free survival (PFS) (e.g., an increase in the duration of PFS experienced by individuals treated according to this method, or an increase in the mean PFS of a population of individuals treated according to this method), increased objective response rate (ORR) (e.g., an increase in the ORR of a population of individuals treated according to this method), and / or increased overall survival (OS) (e.g., an increase in the duration of OS experienced by individuals treated according to this method, or an increase in the mean OS of a population of individuals treated according to this method).

[0348] Increased PFS, ORR, and / or OS can be determined by comparison with, for example, untreated reference individuals and / or reference groups; reference individuals and / or reference groups that have received control therapy (such as one or more previously approved treatments or commercially available products for the treatment of cancer); and / or reference individuals and / or reference groups that have been treated with a PD-1 axis binding antagonist (e.g., atezolizumab) or an anti-TIGIT antagonist antibody (e.g., tislelizumab) as monotherapy. In some aspects, increased PFS, ORR, and / or OS are determined relative to reference individuals and / or reference groups with cancer who have been treated with therapy including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, wherein each individual in the reference individuals and / or reference groups has a TAM characteristic score equal to or lower than a reference TAM characteristic score and / or has an expression level of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO equal to or lower than their respective reference expression levels. The reference TAM feature score described in this paper may be, for example, the median TAM feature score of a reference population of individuals with cancer (e.g., lung cancer, such as NSCLC) or the median expression level of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a reference population of individuals with cancer (e.g., lung cancer, such as NSCLC).

[0349] Those skilled in the art can readily determine whether a given clinical outcome is improved according to the present invention. For example, in this context, "improvement" means that the clinical outcome produced by treating an individual with expression levels of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO higher than their respective reference expression levels, or with a TAM characteristic score higher than a reference TAM characteristic score, by treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., atezolizumab and tireliumab), compared to the clinical outcome produced by the comparative treatment as described above, is at least 3%, at least 5%, at least 7%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 75%, at least 100%, or at least 120% higher.

[0350] For example, in some aspects, the duration of PFS or OS experienced by individuals treated according to the method, or the average PFS or OS of a population of individuals treated according to the method, is increased by at least 3%, at least 5%, at least 7%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 75%, at least 100%, or at least 120%.

[0351] In another instance, in some respects, the ORR of the individual population treated according to this method increased by at least 3%, at least 5%, at least 7%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 75%, at least 100%, or at least 120%.

[0352] Technicians can easily determine the timing for assessing clinical outcomes / endpoints. In principle, the timing is determined at the point when the difference in clinical outcomes / endpoints between the two treatments becomes apparent. This time can 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.

[0353] sample

[0354] The expression levels and / or TAM characteristic scores of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO can be determined from any suitable sample. Exemplary sample types include, but are not limited to, tissue samples, cell samples, whole blood samples, plasma samples, serum samples, and combinations thereof. Samples can be fresh, archived, or frozen.

[0355] In some respects, the sample is a tissue sample, such as a tumor tissue sample. In some respects, the tumor tissue sample is a biopsy section. In some respects where the cancer is lung cancer (e.g., NSCLC), the sample is a biopsy section of lung cancer tissue.

[0356] In some respects, the samples were obtained from the individual prior to treatment with the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody, for example, immediately before the first administration of the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody, or at least one day, at least one week, or at least one month before the first administration of the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody.

[0357] TAM feature scores

[0358] In some respects, determining the TAM characteristic score in a sample from an individual involves calculating the average expression levels of at least two of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual. Therefore, in some respects, the TAM characteristic score is the average (e.g., the average of normalized expression levels) of the expression levels of at least two of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual.

[0359] In some respects, determining the TAM characteristic score in a sample from an individual involves calculating the average expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual. Therefore, in some respects, the TAM characteristic score is the average (e.g., the average normalized expression level) of the expression levels of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual.

[0360] level of expression

[0361] The expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1 and / or MARCO detected in the methods provided herein can be, for example, nucleic acid expression levels or protein expression levels.

[0362] In some respects, expression levels are nucleic acid expression levels, such as mRNA expression levels. Nucleic acid expression levels can be detected using any suitable method known in the art, for example by RNA-seq, quantitative reverse transcriptase PCR (RT-qPCR), quantitative PCR (qPCR), real-time PCR, quantitative real-time PCR (qRT-PCR), multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, in situ hybridization (ISH), or combinations thereof. Other amplification-based methods include, for example, transcription-mediated amplification (TMA), strand substitution amplification (SDA), nucleic acid sequence-based amplification (NASBA), and signal amplification methods such as bDNA.

[0363] In some cases, the nucleic acid expression level of the genes described in this article can be determined using sequencing-based techniques such as RNA-seq, gene expression serial analysis (SAGE), high-throughput sequencing (e.g., massively parallel sequencing), and Sequenom. Nucleic acid expression levels can also be measured using techniques such as NanoString nCounter and High Coverage Expression Profiling (HiCEP). Other protocols for assessing the status of genes and gene products can be found, for example, Ausubel et al., eds., Current Protocols in Molecular Biology, 1995, Units 2 (Northern Blotting), 4 (Southern Blotting), 15 (Immunoblotting), and 18 (PCR Analysis).

[0364] Other methods for detecting the nucleic acid levels of the genes described herein include protocols for examining or detecting mRNA (such as target mRNA) in tissue or cell samples using microarray technology.

[0365] Other methods for detecting the nucleic acid expression levels of the genes described herein include electrophoresis, Northern and Southern blotting, in situ hybridization (e.g., single or multiplex nucleic acid in situ hybridization), RNase protection assays, and microarrays (e.g., Illumina BEADARRAY). TM (Technology); Bead arrays (BADGE) for detecting gene expression.

[0366] In some respects, expression level refers to protein expression level, such as the protein expression level determined by mass spectrometry, Western blotting, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blot, immunoassay methods, surface plasmon resonance, spectroscopy, mass spectrometry, or HPLC.

[0367] Normalization of expression level

[0368] In some respects, the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, and / or MARCO are normalized expression levels; for example, the TAM characteristic score is the average of the normalized expression levels of one or more genes in a sample from an individual.

[0369] In some respects, the TAM characteristic score is the average of the normalized expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in samples from individuals.

[0370] The detected expression levels of genes can be normalized using any of the standard normalization methods known in the art. Those skilled in the art will understand that the normalization method used can depend on the gene expression method employed (e.g., in the case of RT-qPCR, one or more housekeeping genes may be used for normalization, but in the case of RNA-seq, the whole genome or substantially the whole genome may be used as the normalization baseline). For example, the detected expression levels of each gene being measured can be normalized for differences in the amount of one or more genes being measured, variability in the quality of the samples used, and / or variability between assay runs.

[0371] In some cases, normalization can be achieved by detecting the expression of specific one or more normalized genes (including one or more reference genes, such as housekeeping genes (e.g., β-actin)). For example, in some cases, nucleic acid expression levels detected using the methods described herein 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, such as housekeeping genes (e.g., β-actin)). Alternatively, normalization can be based on the mean signal or the median signal of all genes tested. On a gene-by-gene basis, the normalized amount of mRNA measured can be compared to the amount found in reference expression levels. The presence and / or expression level / amount measured in a particular subject sample will be at a percentile within this range, which can be determined by methods well known in the art.

[0372] In other cases, the expression levels of each tested gene were not normalized in order to determine expression levels.

[0373] Any statistical method known in the art can be used to determine the expression level of each gene. For example, the expression level can reflect the median expression level, the median normalized expression level, the average expression level, or the average normalized expression level.

[0374] In some respects, the TAM characteristic score is a numerical value that reflects the aggregated Z-score expression level of the gene combination being measured (e.g., a combination of two or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO, or, for example, a combination of all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO).

[0375] Detection of additional genes

[0376] Any of the methods provided above may further include detecting additional genes in samples from an individual, for example, including detecting the expression levels of at least one of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO, as well as one or more additional genes. In some aspects, the method includes further detecting the expression levels of one or more of the following in samples from an individual: tartrate-resistant acid phosphatase type 5 (ACP5), mast cell-expressed membrane protein 1 (MCEMP1), sterol 27-hydroxylase (CYP27A1), oxidized low-density lipoprotein receptor 1 (OLR1), granulin precursor (GRN), glioma pathogenesis-associated protein 2 (GLIPR2), protein 4 containing a repressor domain (ARRDC4), apolipoprotein E (APOE), folate receptor β (FOLR2), and cathepsin D (CTSD). In some cases, the method includes further detection of the expression levels of one, two, three, four, five, six, seven, eight, nine, or all ten of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD.

[0377] In some respects, determining TAM characteristic scores in samples from individuals includes further testing the expression levels of one, two, three, four, five, six, seven, eight, nine, or all ten of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD.

[0378] For example, in some aspects, the TAM characteristic score is the average (e.g., normalized average) of the expression levels of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, and ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in samples from an individual. The measurement and normalization of expression levels can be performed as described above. For example, in some aspects, the TAM characteristic score is a numerical value of the aggregated Z-score expression level reflecting one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO, and one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD.

[0379] In some aspects, the method includes further detecting the expression levels of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in samples from an individual, and determining a TAM characteristic score from it, wherein the TAM characteristic score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in samples from an individual.

[0380] In this study, individuals were identified as having TAM characteristic scores higher than the reference TAM characteristic score, thereby identifying individuals who could benefit from treatments including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies. The expression levels of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD were detected in samples from the individuals.

[0381] Individuals were identified as having TAM characteristic scores higher than the reference TAM characteristic score, thereby identifying them as individuals who could benefit from treatments including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies. The expression levels of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD were detected in samples from the individuals, and TAM characteristic scores were determined from them. The TAM characteristic score is the average expression level of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in samples from the individuals.

[0382] Reference expression level and TAM feature score

[0383] The terms “reference expression level” and “reference TAM characteristic score” refer to an expression level or TAM characteristic score that is compared with another expression level or TAM characteristic score, for example, to make a diagnostic, predictive, prognostic, and / or treatment decision.

[0384] In some respects, the reference expression level or reference TAM feature score is a pre-specified reference expression level or reference TAM feature score.

[0385] In some respects, the reference expression level or reference TAM characteristic score is the expression level or TAM characteristic score of a reference population (e.g., a population of individuals with cancer, such as a population of individuals with lung cancer (e.g., NSCLC)).

[0386] In some respects, the expression level or TAM characteristic score in the reference population is the median expression level or TAM characteristic score of the reference population.

[0387] In other respects, the expression level or TAM characteristic score in the reference population is the average expression level or TAM characteristic score of the reference population.

[0388] In other respects, the expression level or TAM characteristic score was defined as the 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, and 39th percentiles of the expression level or TAM characteristic score in the reference population. 40th percentile, 41st percentile, 42nd percentile, 43rd percentile, 44th percentile, 45th percentile, 46th percentile, 47th percentile, 48th percentile, 49th percentile, 50th percentile, 51st percentile, 52nd percentile, 53rd percentile, 54th percentile, 55th percentile, 56th percentile, 57th percentile, 58th percentile, 59th percentile 60th percentile, 61st percentile, 62nd percentile, 63rd percentile, 64th percentile, 65th percentile, 66th percentile, 67th percentile, 68th percentile, 69th percentile, 70th percentile, 71st percentile, 72nd percentile, 73rd percentile, 74th percentile, 75th percentile, 76th percentile, 77th percentile, 78th percentile, 79th percentile The 80th, 81st, 82nd, 83rd, 84th, 85th, 86th, 87th, 88th, 89th percentile, 90th, 91st, 92nd, 93rd, 94th, 95th, 96th, 97th, 98th, or 99th percentile.

[0389] In some cases, the reference expression level or reference TAM characteristic score is based on the significant difference in individual responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies between values ​​above, equal to, or below a cutoff value, significantly distinguishing between cutoff values ​​for the first and second subsets of individuals in the same reference population who have been treated with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies (e.g., atezolizumab and tislelizumab). In some respects, an individual's responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies is significantly improved relative to an individual's responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies being equal to or above a cutoff value.

[0390] PD-L1 status

[0391] In some respects, PD-L1 expression levels have been assessed in samples from subjects described herein. In some respects, samples have been identified as having a PD-L1-positive tumor cell fraction (e.g., by immunohistochemistry (IHC) assay, such as by positive staining with an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody is SP263, 22C3, SP142, or 28-8).

[0392] In some respects, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, determined by positive staining with anti-PD-L1 antibody SP263 (e.g., calculated using Ventana SP263 IHC assay).

[0393] In some respects, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with anti-PD-L1 antibody 22C3 (e.g., calculated using pharmDx 22C3 IHC assay).

[0394] Section III(E) provides an exemplary method for assessing PD-L1 expression levels.

[0395] B. Bone marrow markers in serum samples during treatment

[0396] Methods for monitoring response to treatment

[0397] On the other hand, the present invention provides a method for monitoring the response of an individual with cancer (e.g., lung cancer, such as NSCLC) to treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)), the method comprising detecting the expression levels of one or more of the following bone marrow markers (e.g., gene expression levels, ...) in a sample from the individual at time points during or after administration of the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody. For example, protein expression levels or nucleic acid expression levels: macrophage receptor with collagen structure (MARCO), cathepsin inhibitor antimicrobial peptide (CAMP), CD5 antigen-like (CD5L), scavenger receptor cysteine-rich type 1 protein M130 (CD163), neutrophil gelatinase-associated lipotransferase (NGAL), macrophage colony-stimulating factor 1 receptor (CSF1R), CD44 antigen (CD44), apolipoprotein C-II (APOC2), apolipoprotein C-III (APOC3), apolipoprotein C-IV (APOC4), apolipoprotein A-II (APOA2), apolipoprotein C-II, apolipoprotein C-III (APOC3), apolipoprotein C-IV (APOC4), apolipoprotein A-II (APOA2), apolipoprotein C-II (APOC4 ... Protein E (APOE), lactoferrin (TRFL), vascular cell adhesion protein 1 (VCAM1), PERM, β-2-microglobulin (B2MG), LYSC, LYAM1, LCAT, and LIRA3, including one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 (e.g., MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3). Increased expression levels of 63, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or all 20) relative to their respective reference expression levels predict individuals likely to respond to treatment, including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies.

[0398] In some respects, the expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 are increased relative to their respective reference expression levels, thereby predicting that an individual may respond to treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies, and the method further includes administering additional doses of PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies to the individual.

[0399] In some respects, an increase in the expression levels of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 (e.g., gene expression levels, such as protein expression levels or nucleic acid expression levels) is defined as an increase of at least 1.2-fold relative to a reference level (e.g., an increase of 1.3-fold, 1.4-fold, 1.5-fold, 1.6-fold, 1.7-fold, 1.8-fold, 1.9-fold, 2-fold, or more than 2-fold). For example, in some respects, an increase in expression level is between 1.2-fold and 5-fold (e.g., between 1.2-fold and 4-fold, between 1.2-fold and 3-fold, between 1.2-fold and 2-fold, between 1.2-fold and 1.9-fold, between 1.2-fold and 1.7-fold, or between 1.2-fold and 1.5-fold).

[0400] In some respects, the response to treatment is an increase in progression-free survival (PFS) or overall survival (OS).

[0401] In some respects of any of the methods presented in this article, the cancer is lung cancer, such as NSCLC. In some respects, the individual is a person.

[0402] level of expression

[0403] The expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 detected in the methods provided herein can be gene expression levels, such as nucleic acid expression levels or protein expression levels (e.g., protein levels determined by mass spectrometry). Exemplary methods for detecting and normalizing nucleic acid expression levels are provided in Section IIIA above.

[0404] In some contexts, expression level refers to protein expression level, such as that determined by mass spectrometry, Western blotting, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blot, immunoassay methods, surface plasmon resonance, spectroscopy, mass spectrometry, or HPLC. In other contexts, it measures protein expression levels in serum samples.

[0405] In other respects, expression levels are nucleic acid expression levels, such as mRNA expression levels. On one hand, the expression levels of one or more of these are detected in cells derived from an individual's blood sample (e.g., in peripheral blood mononuclear cells (PBMCs) derived from an individual's blood sample).

[0406] sample

[0407] The expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 can be determined from any suitable sample. Exemplary sample types include, but are not limited to, tissue samples, cell samples, whole blood samples, plasma samples, serum samples, and combinations thereof. Samples can be fresh, archived, or frozen.

[0408] In some respects, the sample is a serum sample.

[0409] Samples (e.g., serum samples) can be collected from individuals and can be used to detect the expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 at any appropriate time point following the first administration of treatment to an individual, including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies. For example, treatment can be initiated at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days after starting treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies. Samples are collected for 1 day, 38 days, 39 days, 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 ​​days, 49 days, 50 days, 51 days, 52 days, 53 days, 54 days, 55 days, 56 days, 57 days, 58 days, 59 days, or 60 days (e.g., 1-10 days, 5-15 days, 10-20 days, 15-25 days, 20-30 days, 25-35 days, 30-40 days, 35-45 days, 40-50 days, 45-55 days, or 50-60 days). In other instances, samples were collected approximately one week, two weeks, three weeks, four weeks, five weeks, six weeks, seven weeks, eight weeks, or more than eight weeks after the initiation of treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., samples were collected one, two, three, four, five, six, seven, eight, or more than eight weeks after the initiation of treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody).

[0410] In some respects, the expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 were detected three weeks after the start of treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., samples were collected from individuals three weeks after the start of treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., serum samples), and the expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 were detected therein).

[0411] In some respects, the expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 were detected six weeks after the start of treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., samples were collected from individuals six weeks after the start of treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., serum samples), and the expression levels of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 were detected therein).

[0412] Reference expression level

[0413] The term "reference expression level" refers to an expression level (e.g., protein expression level) that is compared to another expression level, for example, to make diagnostic, predictive, prognostic, and / or treatment decisions.

[0414] In some respects, the reference expression level is the baseline expression level of a sample from an individual at a time point prior to the initiation of treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., a sample collected from the individual immediately before the first administration of the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody, or a sample obtained from the individual at least one day, at least one week, or at least one month prior to the first administration of the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody).

[0415] In some respects, the reference level is a pre-specified reference level.

[0416] In some respects, the reference expression level is the expression level of a reference population (e.g., a population of individuals with cancer, such as a population of individuals with lung cancer (e.g., NSCLC)).

[0417] In some respects, the expression level in the reference group is the median expression level of the reference group.

[0418] In other respects, the expression level in the reference group is the average expression level of the reference group.

[0419] In other respects, expression level was defined as the 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, 39th, 40th, and 41st percentiles of expression level in the reference group. 12th percentile, 42nd percentile, 43rd percentile, 44th percentile, 45th percentile, 46th percentile, 47th percentile, 48th percentile, 49th percentile, 50th percentile, 51st percentile, 52nd percentile, 53rd percentile, 54th percentile, 55th percentile, 56th percentile, 57th percentile, 58th percentile, 59th percentile, 60th percentile 61st percentile, 62nd percentile, 63rd percentile, 64th percentile, 65th percentile, 66th percentile, 67th percentile, 68th percentile, 69th percentile, 70th percentile, 71st percentile, 72nd percentile, 73rd percentile, 74th percentile, 75th percentile, 76th percentile, 77th percentile, 78th percentile, 79th percentile, 80th percentile 100th percentile, 81st percentile, 82nd percentile, 83rd percentile, 84th percentile, 85th percentile, 86th percentile, 87th percentile, 88th percentile, 89th percentile, 90th percentile, 91st percentile, 92nd percentile, 93rd percentile, 94th percentile, 95th percentile, 96th percentile, 97th percentile, 98th percentile, or 99th percentile.

[0420] In some cases, the reference expression level is based on significant differences in individual responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies between values ​​above, equal to, or below a cutoff value, significantly distinguishing between cutoff values ​​for the first and second subsets of individuals in the same reference population who have been treated with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies (e.g., atezolizumab and tislelizumab). In some respects, an individual's responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies is significantly improved relative to an individual's responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies being equal to or above the cutoff value.

[0421] PD-L1 status

[0422] In some respects, PD-L1 expression levels have been assessed in samples from subjects described herein. In some respects, samples have been identified as having a PD-L1-positive tumor cell fraction (e.g., by immunohistochemistry (IHC) assay, such as by positive staining with an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody is SP263, 22C3, SP142, or 28-8).

[0423] In some respects, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, determined by positive staining with anti-PD-L1 antibody SP263 (e.g., calculated using Ventana SP263 IHC assay).

[0424] In some respects, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with anti-PD-L1 antibody 22C3 (e.g., calculated using pharmDx 22C3 IHC assay).

[0425] Section III(E) provides an exemplary method for assessing PD-L1 expression levels.

[0426] C. Regulatory T cell (Treg) genes and characteristics

[0427] Methods for identifying individuals who can benefit from treatment

[0428] (i) Treg gene

[0429] In one aspect, the present invention provides a method for identifying an individual with cancer (e.g., lung cancer, e.g., non-small cell lung cancer (NSCLC)) who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tirelinumab)), the method comprising detecting one or more of the following Treg-related genes (e.g., one, two, three, four, five, or six of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2) in a sample from the individual. The expression levels of the following seven proteins were used to identify individuals who could benefit from treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies: forkhead box protein P3 (FOXP3), cytotoxic T lymphocyte protein 4 (CTLA4), interleukin-10 (IL10), tumor necrosis factor receptor superfamily member 18 (TNFRSF18), CC chemokine receptor type 8 (CCR8), zinc finger protein Eos (IKZF4), and zinc finger protein Helios (IKZF2). Individuals with expression levels of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 above their respective reference expression levels were identified as individuals who could benefit from treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies.

[0430] (ii) Treg characteristic scores

[0431] On the other hand, the present invention provides a method for identifying an individual with cancer (e.g., lung cancer, such as NSCLC) who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)), the method comprising detecting FOXP3, CTLA4, IL10, TNFRSF1 in a sample from the individual. 8. The expression levels of at least two of CCR8, IKZF4, and IKZF2 (e.g., two, three, four, five, six, or all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2) and the determination of regulatory T cell (Treg) characteristic scores from them, wherein Treg characteristic scores higher than a reference Treg characteristic score identify individuals as individuals who may benefit from treatment including PD-1 axis binding antagonists and anti-TIGIT antagonists.

[0432] On the other hand, the present invention provides a method for identifying an individual with cancer (e.g., lung cancer, such as NSCLC) who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tisreliumab)), the method comprising detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual and determining a Treg characteristic score therefrom, wherein a Treg characteristic score higher than a reference Treg characteristic score identifies the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0433] In some aspects, the individual has a Treg eigenvalue score higher than that of a reference Treg eigenvalue score in the sample, and the method further includes administering an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody to the individual. Exemplary methods for determining Treg eigenvalue scores and exemplary reference Treg eigenvalue scores are provided below. Exemplary PD-1 axis-binding antagonists, anti-TIGIT antagonist antibodies, and treatment methods comprising these agents are provided in Section IV.

[0434] In some respects of any of the methods presented in this article, the cancer is lung cancer, such as NSCLC. In some respects, the individual is a person.

[0435] Methods for selecting a therapy

[0436] (i) Treg gene

[0437] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising detecting one or more of the Treg-related genes FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual (e.g., one, two, three, four, five, six, or all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2). The expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2, which are higher than their respective reference expression levels, will identify individuals as those who may benefit from treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies (e.g., PD-1 axis binding antagonists disclosed in Section IV of this document (e.g., atezolizumab) and anti-TIGIT antagonist antibodies disclosed in Section IV of this document (e.g., tislelizumab)).

[0438] (ii) Treg characteristic scores

[0439] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising detecting the expression levels of at least two of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 (e.g., one, two, three, four, five, six, or all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2) in a sample from the individual and determining a Treg characteristic score therefrom, wherein a Treg characteristic score higher than a reference Treg characteristic score identifies the individual as an individual who may benefit from treatment including a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0440] On the other hand, the present invention provides a method for selecting a therapy for an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual and determining a Treg eigenvalue score therefrom, wherein a Treg eigenvalue score higher than a reference Treg eigenvalue score identifies the individual as an individual who may benefit from treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0441] In some respects, the individual has a Treg characteristic score higher than that of a reference Treg characteristic score in the sample, and the method further includes administering an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody to the individual.

[0442] Treatment

[0443] (i) Treg gene

[0444] On the other hand, the present invention provides a method for treating an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising (a) detecting the expression levels of one or more of the Treg-related genes FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 (e.g., one, two, three, four, five, six, or all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2) in a sample from the individual, wherein FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2, are present in a sample from the individual. The expression levels of one or more of A4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 are higher than their respective reference expression levels, thereby identifying the individual as an individual who may benefit from treatment including PD-1 axis-binding antagonists and anti-TIGIT antagonist antibodies; and (b) administering to the individual an effective amount of PD-1 axis-binding antagonists and anti-TIGIT antagonist antibodies (e.g., PD-1 axis-binding antagonists disclosed in Section IV of this document (e.g., atezolizumab) and anti-TIGIT antagonist antibodies disclosed in Section IV of this document (e.g., tislelizumab)).

[0445] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tisreliumab)), wherein the individual has been determined to have expression levels of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 above their respective reference expression levels, thereby identifying the individual as an individual who may benefit from treatment including the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

[0446] (ii) Treg characteristic scores

[0447] On the other hand, the present invention provides a method for treating an individual suffering from cancer (e.g., lung cancer, such as NSCLC), the method comprising (a) detecting the expression levels of at least two of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 (e.g., two, three, four, five, six, or all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2) in a sample from the individual and from... It determines a Treg eigenvalue, wherein the Treg eigenvalue is higher than a reference Treg eigenvalue, and thereby identifies the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody; and (b) administers to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0448] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising (a) detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual and determining a Treg characteristic score therefrom, wherein the Treg characteristic score is higher than a reference Treg characteristic score, and thereby identifying the individual as an individual who may benefit from treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody; and (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tislelizumab)).

[0449] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tisreliumab)), wherein the individual has been determined to have a Treg eigenvalue score higher than a reference Treg eigenvalue score, thereby identifying the individual as Individuals who may benefit from treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies, wherein the Treg characteristic score is based on the expression level of at least two of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 (e.g., two, three, four, five, six or all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2) detected in samples from the individual.

[0450] On the other hand, the present invention provides a method for treating an individual with cancer (e.g., lung cancer, such as NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., the PD-1 axis-binding antagonist disclosed in Section IV of this document (e.g., atezolizumab) and the anti-TIGIT antagonist antibody disclosed in Section IV of this document (e.g., tisreliumab)), wherein the individual has been determined to have a Treg eigenvalue score higher than a reference Treg eigenvalue score, thereby identifying the individual as an individual who may benefit from treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, and wherein the Treg eigenvalue score is based on the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 detected in a sample from the individual.

[0451] Benefit

[0452] In some respects, the benefits achieved through treatment including PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies are increased progression-free survival (PFS) (e.g., an increase in the duration of PFS experienced by individuals treated according to this method, or an increase in the mean PFS of a population of individuals treated according to this method), increased objective response rate (ORR) (e.g., an increase in the ORR of a population of individuals treated according to this method), and / or increased overall survival (OS) (e.g., an increase in the duration of OS experienced by individuals treated according to this method, or an increase in the mean OS of a population of individuals treated according to this method).

[0453] Increased PFS, ORR, and / or OS can be determined by comparison with, for example, untreated reference individuals and / or reference groups; reference individuals and / or reference groups that have received control therapy (such as one or more previously approved treatments or commercially available products for the treatment of cancer); and / or reference individuals and / or reference groups that have been treated with a PD-1 axis binding antagonist (e.g., atezolizumab) or an anti-TIGIT antagonist antibody (e.g., tislelizumab) as monotherapy. In some respects, the increased PFS, ORR, and / or OS are determined relative to reference individuals and / or reference groups of individuals with cancer who have been treated with therapy including PD-1 axis-binding antagonists and anti-TIGIT antagonist antibodies, wherein each individual in the reference individuals and / or reference groups has a Treg eigenvalue equal to or lower than a reference Treg eigenvalue and / or has an expression level of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 equal to or lower than their respective reference expression levels. The reference Treg eigenvalue is described herein and may be, for example, the median Treg eigenvalue of a reference group of individuals with cancer (e.g., lung cancer, such as NSCLC) or the median expression level of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a reference group of individuals with cancer (e.g., lung cancer, such as NSCLC).

[0454] Section III(A) provides an exemplary method for determining whether a given clinical outcome is improved according to the invention.

[0455] sample

[0456] The expression levels of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2, and / or Treg characteristic scores, can be determined from any suitable sample. Exemplary sample types include, but are not limited to, tissue samples, cell samples, whole blood samples, plasma samples, serum samples, and combinations thereof. Samples can be fresh, archived, or frozen.

[0457] In some respects, the sample is a tissue sample, such as a tumor tissue sample. In some respects, the tumor tissue sample is a biopsy section. In some respects where the cancer is lung cancer (e.g., NSCLC), the sample is a biopsy section of lung cancer tissue.

[0458] In some respects, the samples were obtained from the individual prior to treatment with the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody, for example, immediately before the first administration of the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody, or at least one day, at least one week, or at least one month before the first administration of the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody.

[0459] Treg feature scores

[0460] In some aspects, determining the Treg characteristic score in a sample from an individual involves calculating the average expression levels of at least two of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in the sample from the individual. Therefore, in some aspects, the Treg characteristic score is the average (e.g., the average of normalized expression levels) of the expression levels of at least two of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in the sample from the individual.

[0461] In some respects, determining the Treg characteristic score in a sample from an individual involves calculating the average expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in the sample from the individual. Therefore, in some respects, the TAM characteristic score is the average (e.g., the average normalized expression level) of the expression levels of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in the sample from the individual.

[0462] level of expression

[0463] The expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and / or IKZF2 detected in the methods provided herein can be, for example, nucleic acid expression levels or protein expression levels (e.g., protein levels determined by mass spectrometry).

[0464] In some respects, expression levels are nucleic acid expression levels, such as mRNA expression levels. Nucleic acid expression levels can be detected using any suitable method known in the art, for example by RNA-seq, RT-qPCR, qPCR, real-time PCR, quantitative real-time PCR (qRT-PCR), multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, in situ hybridization (ISH), or combinations thereof. Section III(A) provides further exemplary methods for measuring nucleic acid expression levels.

[0465] In some respects, expression level refers to protein expression level, such as the protein expression level determined by mass spectrometry, Western blotting, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blot, immunoassay methods, surface plasmon resonance, spectroscopy, mass spectrometry, or HPLC.

[0466] Normalization of expression level

[0467] In some respects, the expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and / or IKZF2 are normalized expression levels; for example, the Treg eigenvalue is the average of the normalized expression levels of one or more genes in a sample from an individual.

[0468] In some respects, the Treg characteristic score is the average of the normalized expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in samples from an individual.

[0469] Section III(A) provides an exemplary method for normalizing gene expression levels.

[0470] In some respects, the Treg characteristic score is a numerical value that reflects the aggregated Z-score expression level of a combination of genes being measured (e.g., a combination of two or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2, or a combination of all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2).

[0471] Reference expression level and Treg feature score

[0472] The terms “reference expression level” and “reference Treg characteristic score” refer to an expression level or Treg characteristic score that is compared with another expression level or Treg characteristic score, for example, to make a diagnostic, predictive, prognostic, and / or treatment decision.

[0473] In some respects, the reference expression level or reference Treg feature score is a pre-specified reference expression level or reference Treg feature score.

[0474] In some respects, the reference expression level or reference Treg characteristic score is the expression level or Treg characteristic score of a reference population (e.g., a population of individuals with cancer, such as a population of individuals with lung cancer (e.g., NSCLC)).

[0475] In some respects, the expression level or Treg characteristic score in the reference population is the median expression level or Treg characteristic score of the reference population.

[0476] In other respects, the expression level or Treg characteristic score in the reference population is the average expression level or Treg characteristic score of the reference population.

[0477] In other respects, the expression level or Treg characteristic score was defined as the 25th, 26th, 27th, 28th, 29th, 30th, 31st, 32nd, 33rd, 34th, 35th, 36th, 37th, 38th, and 39th percentiles of the expression level or Treg characteristic score in the reference population. Number, 40th percentile, 41st percentile, 42nd percentile, 43rd percentile, 44th percentile, 45th percentile, 46th percentile, 47th percentile, 48th percentile, 49th percentile, 50th percentile, 51st percentile, 52nd percentile, 53rd percentile, 54th percentile, 55th percentile, 56th percentile, 57th percentile, 58th percentile, 59th percentile 60th percentile, 61st percentile, 62nd percentile, 63rd percentile, 64th percentile, 65th percentile, 66th percentile, 67th percentile, 68th percentile, 69th percentile, 70th percentile, 71st percentile, 72nd percentile, 73rd percentile, 74th percentile, 75th percentile, 76th percentile, 77th percentile, 78th percentile, 79th percentile The 80th percentile, 81st percentile, 82nd percentile, 83rd percentile, 84th percentile, 85th percentile, 86th percentile, 87th percentile, 88th percentile, 89th percentile, 90th percentile, 91st percentile, 92nd percentile, 93rd percentile, 94th percentile, 95th percentile, 96th percentile, 97th percentile, 98th percentile, or 99th percentile.

[0478] In some cases, the reference expression level or reference Treg characteristic score is based on significant differences in individual responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies between values ​​above, equal to, or below a cutoff value, significantly distinguishing between cutoff values ​​for the first and second subsets of individuals in the same reference population who have been treated with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies (e.g., atezolizumab and tislelizumab). In some respects, an individual's responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies is significantly improved relative to an individual's responsiveness to treatment with PD-1 axis binding antagonists and anti-TIGIT antagonist antibodies being equal to or above the cutoff value.

[0479] PD-L1 status

[0480] In some respects, PD-L1 expression levels have been assessed in samples from subjects described herein. In some respects, samples have been identified as having a PD-L1-positive tumor cell fraction (e.g., by immunohistochemistry (IHC) assay, such as by positive staining with an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody is SP263, 22C3, SP142, or 28-8).

[0481] In some respects, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, determined by positive staining with anti-PD-L1 antibody SP263 (e.g., calculated using Ventana SP263 IHC assay).

[0482] In some respects, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with anti-PD-L1 antibody 22C3 (e.g., calculated using pharmDx 22C3 IHC assay).

[0483] Section III(E) provides an exemplary method for assessing PD-L1 expression levels.

[0484] D. TIGIT expression assessment

[0485] In some respects, TIGIT expression in the individuals described herein is assessed. The methods provided herein may include determining the level of TIGIT expression in a biological sample (e.g., a tumor sample) obtained from the individual. In other instances, the level of TIGIT expression in a biological sample (e.g., a tumor sample) obtained from the individual has been determined before or after the initiation of treatment. TIGIT expression can be determined using any suitable method. Any suitable tumor sample can be used, such as formalin-fixed and paraffin-embedded (FFPE) tumor samples, archived tumor samples, fresh tumor samples, or frozen tumor samples.

[0486] For example, TIGIT expression can be determined as the percentage of a tumor sample occupied by tumor-infiltrating immune cells expressing a detectable level of TIGIT, as the percentage of tumor-infiltrating immune cells expressing a detectable level of TIGIT in the tumor sample, and / or as the percentage of tumor cells expressing a detectable level of TIGIT in the tumor sample. It should be understood that in any of the foregoing examples, the percentage of a tumor sample occupied by tumor-infiltrating immune cells can be the percentage of the tumor area covered by tumor-infiltrating immune cells in a slice of a tumor sample obtained from an individual, for example, as assessed by IHC using an anti-TIGIT antagonist antibody. Any suitable anti-TIGIT antagonist antibody can be used. In some examples, the anti-TIGIT antagonist antibody is 10A7 (WO 2009 / 126688A3; US Patent No.: 9,499,596). In other examples, the anti-TIGIT antagonist antibody is the anti-human TIGIT rabbit monoclonal antibody clone SP410 (Roche Tissue Diagnostics, Pleasanton, CA). In some respects, the VENTANA OptiView DAB IHC assay kit was used to detect TIGIT antagonist antibodies (e.g., SP410) on the automated VENTANA BenchMark ULTRA platform.

[0487] Assessment of E.PD-L1 expression

[0488] In some aspects, PD-L1 expression in the individuals described herein is assessed. The methods provided herein may include determining the level of PD-L1 expression in a biological sample (e.g., a tumor sample) obtained from the individual. In other examples, the level of PD-L1 expression in a biological sample (e.g., a tumor sample) obtained from the individual has been determined before or after the initiation of treatment. PD-L1 expression can be determined using any suitable method. For example, PD-L1 expression can be determined as described in U.S. Patent Application Publications US20180030138A1 and US20180037655A1. Any suitable tumor sample can be used, such as formalin-fixed and paraffin-embedded (FFPE) tumor samples, archived tumor samples, fresh tumor samples, or frozen tumor samples.

[0489] For example, PD-L1 expression can be determined as the percentage of tumor-infiltrating immune cells expressing a detectable level of PD-L1 in a tumor sample, as the percentage of tumor-infiltrating immune cells expressing a detectable level of PD-L1 in the tumor sample, and / or as the percentage of tumor cells expressing a detectable level of PD-L1 in the tumor sample. It should be understood that in any of the foregoing examples, the percentage of tumor-infiltrating immune cells in a tumor sample can be the percentage of the tumor area covered by tumor-infiltrating immune cells in a slice of a tumor sample obtained from an individual, for example, as assessed by IHC using an anti-PD-L1 antibody (e.g., SP142 antibody). Any suitable anti-PD-L1 antibody can be used, including, for example, SP142 (Ventana), SP263 (Ventana), 22C3 (Dako), 28-8 (Dako), E1L3N (CellSignaling Technology), 4059 (ProSci, Inc.), h5H1 (Advanced Cell Diagnostics), and 9A11. In some examples, the anti-PD-L1 antibody is SP142. In other examples, the anti-PD-L1 antibody is SP263. In some examples, the anti-PD-L1 antibody is 22C3. In some instances, the anti-PD-L1 antibody is 28-8.

[0490] In some instances, tumor samples obtained from individuals showed detectable PD-L1 expression levels in less than 1% of tumor cells, 1% or more of tumor cells, 1% to less than 5% of tumor cells, 5% or more of tumor cells, 5% to less than 50% of tumor cells, or 50% or more of tumor cells.

[0491] In some instances, tumor samples obtained from individuals showed detectable PD-L1 expression levels in tumor-infiltrating immune cells that comprised less than 1% of the tumor sample, more than 1% of the tumor sample, 1% to less than 5% of the tumor sample, more than 5% of the tumor sample, 5% to less than 10% of the tumor sample, or more than 10% of the tumor sample.

[0492] In some aspects, tumor samples obtained from an individual have detectable PD-L1 expression levels in tumor-infiltrating immune cells comprising 5% to 19% (e.g., TIC 5% to 19%) of the tumor sample; for example, having low PD-L1 expression levels. In some aspects, tumor samples obtained from an individual have detectable PD-L1 expression levels in tumor-infiltrating immune cells comprising ≥20% (e.g., TIC ≥20%) of the tumor sample; for example, having high PD-L1 expression levels. In some embodiments, tumor samples with a TIC greater than or equal to 5% have been identified as equivalent to CPS greater than or equal to 1.

[0493] In some embodiments, tumor samples can be scored based on PD-L1 positivity in tumor-infiltrating immune cells and / or tumor cells according to the diagnostic assessment criteria shown in Table 1 and / or Table 2, respectively.

[0494] Table 1. Diagnostic criteria for tumor-infiltrating immune cells (IC) using IHC

[0495]

[0496]

[0497] Table 2. Diagnostic criteria for tumor cell (TC) IHC

[0498]

[0499] In some cases, in any of the methods, uses, or compositions described herein, the individual has a PD-L1-selective tumor (e.g., a tumor sample in which the proportion of the tumor area occupied by PD-L1-expressing tumor-infiltrating immune cells (ICs) is greater than or equal to 5%, as determined by IHC using an SP142 antibody). In some cases, a PD-L1-selective tumor is a tumor in which the proportion of the tumor area occupied by PD-L1-expressing immune cells (ICs) is greater than or equal to 5%, as determined by immunohistochemical (IHC) assay. In some cases, the IHC assay uses anti-PD-L1 antibodies SP142, SP263, 22C3, or 28-8. In some cases, the IHC assay uses anti-PD-L1 antibody SP142. In some cases, the IHC assay uses anti-PD-L1 antibody SP263. In some cases, the IHC assay uses anti-PD-L1 antibody 22C3. In some cases, the IHC assay uses anti-PD-L1 antibody 22C3. In some cases, the IHC assay uses anti-PD-L1 antibody 28-8.

[0500] In some cases, the IC score has been determined to be greater than or equal to 5% (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some cases, the IC score has been determined to be 2 or 3 (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some cases, the IC score has been determined to be greater than or equal to 1% (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some cases, the IC score has been determined to be greater than or equal to 10% (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some cases, the IC score has been determined to be greater than or equal to 1% and less than 50% (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some cases, the IC score has been determined to be greater than or equal to 1% and less than 30% (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay).

[0501] In some cases, in any of the methods, uses, or compositions described herein, tumor samples obtained from an individual have detectable levels of PD-L1 protein expression. In some cases, the detectable PD-L1 protein expression level has been determined by IHC assay. In some cases, the IHC assay uses the anti-PD-L1 antibody SP142. In some cases, a tumor sample has been identified as having detectable PD-L1 expression levels in ≥5% of the tumor-infiltrating immune cells. In some cases, a tumor sample has been identified as having detectable PD-L1 expression levels in ≥1% of the tumor-infiltrating immune cells. In some cases, a tumor sample has been identified as having detectable PD-L1 expression levels in ≥1% and <5% of the tumor-infiltrating immune cells. In some cases, a tumor sample has been identified as having detectable PD-L1 expression levels in ≥5% and <10% of the tumor-infiltrating immune cells. In some cases, a tumor sample is identified as having detectable PD-L1 expression levels in ≥10% of the tumor-infiltrating immune cells. In some cases, a tumor sample is identified as having detectable PD-L1 expression levels in ≥1% of the tumor cells. In some cases, a tumor sample is identified as having detectable PD-L1 expression levels in ≥1% and <5% of the tumor cells. In some cases, a tumor sample is identified as having detectable PD-L1 expression levels in ≥5% and <50% of the tumor cells. In some cases, a tumor sample is identified as having detectable PD-L1 expression levels in ≥50% of the tumor cells.

[0502] In some cases, in any of the methods, uses, or compositions described herein, the individual has a PD-L1-selective tumor (e.g., a PD-L1 "high"-selective tumor (e.g., a PD-L1 tumor proportion fraction (TPS) greater than or equal to 50% in a tumor sample, determined by using an IHC antibody against SP263). In some cases, the PD-L1-selective tumor is a PD-L1 "high"-selective tumor. In some cases, the PD-L1-selective tumor is a tumor that has been identified by immunohistochemical (IHC) assay. The TPS is greater than or equal to 50%. In some cases, the IHC assay uses anti-PD-L1 antibodies SP263, SP142, 22C3, or 28-8. In some cases, the IHC assay uses anti-PD-L1 antibody SP263. In some cases, the IHC assay uses anti-PD-L1 antibody SP142. In some cases, the IHC assay uses anti-PD-L1 antibody 22C3. In some cases, the TPS has been determined to be greater than or equal to 50% (e.g., as with Ventana (SP263) PD-L1). In some cases, TPS has been determined to be less than 50% (e.g., as determined using the Ventana (SP263) PD-L1 IHC assay). In some cases, TPS has been determined to be greater than or equal to 1% (e.g., as determined using the Ventana (SP263) PD-L1 IHC assay). In some cases, TPS has been determined to be greater than or equal to 1% and less than 50% (e.g., as determined using the Ventana (SP263) PD-L1 IHC assay).

[0503] In some cases, in any of the methods, uses, or compositions described herein, tumor samples obtained from an individual have detectable levels of PD-L1 protein expression. In some cases, the detectable PD-L1 protein expression level has been determined by IHC assay. In some cases, the IHC assay uses the anti-PD-L1 antibody SP263. In some cases, the tumor sample has been determined to have a PD-L1-positive tumor cell fraction greater than or equal to 50% of the tumor sample. In some cases, the tumor sample has been determined to have a PD-L1-positive tumor cell fraction less than 50% of the tumor sample. In some cases, the tumor sample has been determined to have a PD-L1-positive tumor cell fraction greater than or equal to 1% and less than 50% of the tumor sample.

[0504] In some cases, the IHC assay uses anti-PD-L1 antibody 22C3. In some cases, the IHC assay is the pharmDx 22C3 IHC assay. In some cases, the PD-L1 positive tumor cell fraction is greater than or equal to 50%, determined by positive staining with anti-PD-L1 antibody 22C3. In some embodiments, a tumor sample has been determined to have a composite positive score (CPS) greater than or equal to 10 or a tumor proportion score (TPS) greater than or equal to 1% of the tumor sample, for example, as determined by using anti-PD-L1 antibody 22C3 as part of the pharmDx 22C3 IHC assay. In some embodiments, a tumor sample has been determined to have a CPS greater than or equal to 10 or a TPS greater than or equal to 1% and less than 50% of the tumor sample, for example, as determined by using anti-PD-L1 antibody 22C3 as part of the pharmDx 22C3 IHC assay. In some embodiments, a tumor sample has been determined to have a CPS greater than or equal to 20 or a TPS greater than or equal to 50% in the tumor sample, for example, as determined by using anti-PD-L1 antibody 22C3 as part of the pharmDx 22C3 IHC assay. In some embodiments, a tumor sample having a CPS greater than or equal to 1 has been determined to be equivalent to a TIC greater than or equal to 5%.

[0505] In some cases, the IHC assay uses anti-PD-L1 antibody 28-8. In other cases, the IHC assay is the pharmDx 28-8 IHC assay. In some cases, the fraction of PD-L1 positive tumor cells is greater than or equal to 50%, determined by positive staining with anti-PD-L1 antibody 28-8.

[0506] In some cases, in any of the methods, uses, or compositions described herein, tumor samples obtained from an individual have detectable levels of PD-L1 nucleic acid expression. In some cases, the detectable PD-L1 nucleic acid expression level has been determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or combinations thereof. In some cases, the sample is selected from the group consisting of tissue samples, whole blood samples, serum samples, and plasma samples. In some cases, the tissue sample is a tumor sample. In some cases, the tumor sample contains infiltrating tumor immune cells, tumor cells, stromal cells, and any combination thereof.

[0507] IV. Exemplary anti-TIGIT antagonist antibodies and PD-1 axis binding antagonists

[0508] This document describes exemplary anti-TIGIT antagonist antibodies and PD-1 axis binding antagonists of the methods, uses and compositions provided for use according to the present invention, which may be used to treat individuals (e.g., humans) with cancer.

[0509] A. Exemplary anti-TIGIT antagonist antibody

[0510] The present invention provides an anti-TIGIT antagonist antibody that can be used to treat cancer in subjects (e.g., humans).

[0511] In some cases, the anti-TIGIT antagonist antibody is tisrelimumab (CAS Registry No.: 1918185-84-8). Tirelimumab (Genentech) is also known as MTIG7192A.

[0512] In certain circumstances, anti-TIGIT antagonist antibodies comprise at least one, two, three, four, five, or six HVRs selected from the following: (a) HVR-H1 containing the amino acid sequence of SNSAAWN (SEQ ID NO:1); (b) HVR-H2 containing the amino acid sequence of KTYYRFKWYSDYAVSVKG (SEQ ID NO:2); (c) HVR-H3 containing the amino acid sequence of ESTTYDLLAGPFDY (SEQ ID NO:3); (d) HVR-L1 containing the amino acid sequence of KSSQTVLYSSNNKKYLA (SEQ ID NO:4); (e) HVR-L2 containing the amino acid sequence of WASTRES (SEQ ID NO:5); and / or (f) HVR-L3 containing the amino acid sequence of QQYYSTPFT (SEQ ID NO:6), or one or more of the above HVRs and HVRs containing the amino acid sequence of SEQ ID NO:6. Any combination of one or more variants of IDNO:1-6 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity).

[0513] In some cases, anti-TIGIT antagonist antibodies may include: (a) HVR-H1, which contains the amino acid sequence of SNSAAWN (SEQ ID NO: 1); (b) HVR-H2, which contains

[0514] The amino acid sequence of KTYYRFKWYSDYAVSVKG (SEQ ID NO:2); (c) HVR-H3, which contains the amino acid sequence of ESTTYDLLAGPFDY (SEQ ID NO:3); (d) HVR-L1, which contains the amino acid sequence of KSSQTVLYSSNNKKYLA (SEQ ID NO:4); (e) HVR-L2, which contains the amino acid sequence of WASTRES (SEQ ID NO:5); and (f) HVR-L3, which contains the amino acid sequence of QQYYSTPFT (SEQ ID NO:6). In some cases, anti-TIGIT antagonist antibodies have: a VH domain containing the amino acid sequence of KTYYRFKWYSDYAVSVKG (SEQ ID NO:2); (c) HVR-H3, which contains the amino acid sequence of KTYYRFKWYSDYAVSVKG (SEQ ID NO:3); (d) HVR-L1, which contains the amino acid sequence of KSSQTVLYSSNNKKYLA (SEQ ID NO:4); (e) HVR-L2, which contains the amino acid sequence of WASTRES (SEQ ID NO:5); and (f) HVR-L3, which contains the amino acid sequence of QQYYSTPFT (SEQ ID NO:6).

[0515] The amino acid sequence EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO:17) has at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) or the sequence of SEQ ID NO:17, or is identical to QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO:17). NO:18) has an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) or the sequence of SEQ ID NO:18; and / or a VL domain containing an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity); ...

[0516] DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAWYQQKPGQPPNLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPFTFGPGTKVEIK (SEQ ID NO:19) has an amino acid sequence with at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) or the sequence of SEQ ID NO:19. In some cases, the anti-TIGIT antagonist antibody has: a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with sequence SEQ ID NO:17, or the sequence of SEQ ID NO:17; and / or a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with sequence SEQ ID NO:19, or the sequence of SEQ ID NO:19. In some cases, the anti-TIGIT antagonist antibody has: a VH domain comprising the amino acid sequence of SEQ ID NO:17; and a VL domain comprising the amino acid sequence of SEQ ID NO:19. In some cases, the anti-TIGIT antagonist antibody has: a VH domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with sequence SEQ ID NO:18, or the sequence of SEQ ID NO:18; and / or a VL domain comprising an amino acid sequence having at least 90% sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) with sequence SEQ ID NO:19, or the sequence of SEQ ID NO:19. In some cases, the anti-TIGIT antagonist antibody has: a VH domain comprising the amino acid sequence of SEQ ID NO:18; and a VL domain comprising the amino acid sequence of SEQ ID NO:19.

[0517] In some cases, anti-TIGIT antagonist antibodies comprise both heavy and light chain sequences, wherein: (a) the heavy chain contains the following amino acid sequence:

[0518] EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWL

[0519] GKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCT

[0520] RESTTYDLLAGPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAA

[0521] LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS

[0522] SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF

[0523] LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK

[0524] PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA

[0525] KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN

[0526] NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(SEQ ID NO: 33); and (b) the light chain contains the following amino acid sequence: DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAWYQQKPGQPPNLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYST PFTFGPGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:34). In some respects, the anti-TIGIT antagonist antibody comprises: (a) a heavy chain containing the amino acid sequence of SEQ ID NO:33, and (b) a light chain containing the amino acid sequence of SEQ ID NO:34.

[0527] In some cases, the anti-TIGIT antagonist antibody further comprises at least one, two, three, or four of the following light chain variable region (FR) framework regions: FR-L1, which contains the amino acid sequence DIVMTQSPDSLAVSLGERATINC (SEQ ID NO:7); FR-L2, which contains the amino acid sequence WYQQKPGQPPNLLIY (SEQ ID NO:8); FR-L3, which contains the amino acid sequence GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO:9); and / or FR-L4, which contains the amino acid sequence FGPGTKVEIK (SEQ ID NO:10), or a combination of one or more of the above FRs with one or more variants thereof, which have at least about 90% sequence identity with any of SEQ ID NO:7 to 10 (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). In some cases, the antibody further comprises, for example: FR-L1, which contains the amino acid sequence DIVMTQSPDSLAVSLGERATINC (SEQ ID NO:7); FR-L2, which contains the amino acid sequence WYQQKPGQPPNLLIY (SEQ ID NO:8); FR-L3, which contains the amino acid sequence GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO:9); and FR-L4, which contains the amino acid sequence FGPGTKVEIK (SEQ ID NO:10).

[0528] In some cases, anti-TIGIT antagonist antibodies further comprise at least one, two, three, or four of the following heavy chain variable regions FR: FR-H1, which contains

[0529] The amino acid sequence X1VQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO:11), where X1 is E or Q; FR-H2, which contains the amino acid sequence WIRQSPSRGLEWLG (SEQ ID NO:12); FR-H3, which contains

[0530] The amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO:13); and / or FR-H4, which contains the amino acid sequence of WGQGTLVTVSS (SEQ ID NO:14), or a combination of one or more of the above FRs with one or more variants thereof, which one or more variants thereof having at least about 90% sequence identity with any of SEQ ID NO:11 to 14 (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity). The anti-TIGIT antagonist antibody may further comprise, for example, at least one, two, three, or four of the following heavy chain variable regions FR: FR-H1, which contains

[0531] The amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO:15); FR-H2, which contains the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO:12); FR-H3, which contains the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO:13); and / or FR-H4, which contains the amino acid sequence of WGQGTLVTVSS (SEQ ID NO:14), or a combination of one or more of the above FRs with one or more variants thereof, which have at least about 90% sequence identity with any of SEQ ID NO:12 to 15 (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity). In some cases, anti-TIGIT antagonist antibodies include: FR-H1, which contains the amino acid sequence EVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO:15); FR-H2, which contains the amino acid sequence WIRQSPSRGLEWLG (SEQ ID NO:12); FR-H3, which contains

[0532] The amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO:13); and FR-H4, which contains the amino acid sequence of WGQGTLVTVSS (SEQ ID NO:14). In another case, for example, the anti-TIGIT antagonist antibody may further comprise at least one, two, three, or four of the following heavy chain variable regions FR: FR-H1, which contains

[0533] The amino acid sequence of QVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO:16); FR-H2, which contains the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO:12); FR-H3, which contains the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO:13); and / or FR-H4, which contains the amino acid sequence of WGQGTLVTVSS (SEQ ID NO:14), or a combination of one or more of the above FRs with one or more variants thereof, which have at least about 90% sequence identity with any of SEQ ID NO:12 to 14 and 16 (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity). In some cases, anti-TIGIT antagonist antibodies include: FR-H1, which contains the amino acid sequence QVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO:16); FR-H2, which contains the amino acid sequence WIRQSPSRGLEWLG (SEQ ID NO:12); FR-H3, which contains

[0534] The amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO:13); and FR-H4, which contains the amino acid sequence of WGQGTLVTVSS (SEQ ID NO:14).

[0535] On the other hand, an anti-TIGIT antagonist antibody is provided, wherein the antibody comprises VH as provided above and VL as provided above, wherein one or both variable domain sequences include post-translational modifications.

[0536] In some cases, any of the aforementioned anti-TIGIT antagonist antibodies can bind to both human TIGIT and rabbit TIGIT. In some cases, any of the aforementioned anti-TIGIT antagonist antibodies can bind to both human TIGIT and cynomolgus monkey (cynomolgus monkey) TIGIT. In some cases, any of the aforementioned anti-TIGIT antagonist antibodies can bind to human TIGIT, cyno TIGIT, and rabbit TIGIT. In some cases, any of the aforementioned anti-TIGIT antagonist antibodies can bind to human TIGIT, cyno TIGIT, and rabbit TIGIT, but not to mouse TIGIT.

[0537] In some cases, anti-TIGIT antagonist antibodies are used at approximately 10 nM or lower K+. DCombined with human TIGIT and at approximately 10 nM or lower K D Combined with cyno TIGIT (e.g., K at approximately 0.1 nM to approximately 1 nM) D It binds to human TIGIT at a concentration of approximately 0.5 nM to approximately 1 nM K. D Combined with cyno TIGIT, for example at approximately 0.1 nM or lower K D It binds to human TIGIT at a concentration of approximately 0.5 nM or lower. D (Combined with Cyno TIGIT).

[0538] In some cases, anti-TIGIT antagonist antibodies specifically bind to TIGIT and inhibit or block the interaction between TIGIT and the poliovirus receptor (PVR) (e.g., the antagonist antibody inhibits intracellular signaling mediated by TIGIT-PVR binding). In some cases, the antagonist antibody inhibits or blocks the binding of human TIGIT to human PVR with an IC50 value of 10 nM or less (e.g., 1 nM to about 10 nM). In some cases, anti-TIGIT antagonist antibodies specifically bind to TIGIT and inhibit or block the interaction between TIGIT and PVR without affecting the PVR-CD226 interaction. In some cases, the antagonist antibody inhibits or blocks the binding of cyno TIGIT to cyno PVR with an IC50 value of 50 nM or less (e.g., 1 nM to about 50 nM, such as 1 nM to about 5 nM). In some cases, anti-TIGIT antagonist antibodies inhibit and / or block the interaction between CD226 and TIGIT. In some cases, anti-TIGIT antagonist antibodies inhibit and / or block the ability of TIGIT to disrupt CD226 homodimerization.

[0539] In some cases, the methods or uses described herein may include the use or administration of isolated anti-TIGIT antagonist antibodies that compete with any of the anti-TIGIT antagonist antibodies described above for binding to TIGIT. For example, the method may include administering an isolated anti-TIGIT antagonist antibody that competitively binds to TIGIT with an anti-TIGIT antagonist antibody having the following six HVRs: (a) HVR-H1, which contains the amino acid sequence SNSAAWN (SEQ ID NO:1); (b) HVR-H2, which contains the amino acid sequence KTYYRFKWYSDYAVSVKG (SEQ ID NO:2); (c) HVR-H3, which contains the amino acid sequence ESTTYDLLAGPFDY (SEQ ID NO:3); (d) HVR-L1, which contains the amino acid sequence KSSQTVLYSSNNKKYLA (SEQ ID NO:4); (e) HVR-L2, which contains the amino acid sequence WASTRES (SEQ ID NO:5); and (f) HVR-L3, which contains the amino acid sequence QQYYSTPFT (SEQ ID NO:6). The method described herein may also include administering an isolated anti-TIGIT antagonist antibody that binds to the same epitope as the aforementioned anti-TIGIT antagonist antibody.

[0540] In some respects, anti-TIGIT antagonist antibodies are antibodies with full Fc-mediated effector function (e.g., tirelimumab, vimbrolizumab, atelimumab, EOS084448, or TJ-T6) or enhanced effector function (e.g., SGN-TGT).

[0541] In some aspects, anti-TIGIT antagonist antibodies contain an Fc domain capable of interacting with (e.g., activating) the Fcγ receptor (FcγR). In other aspects, anti-TIGIT antagonist antibodies contain an Fc domain capable of interacting with (e.g., activating) the FcγR of one or more bone marrow cell types (e.g., cDC1, macrophages, neutrophils, and circulating monocytes).

[0542] In some respects, anti-TIGIT antagonist antibodies can Fc-dependently activate one or more bone marrow cell types, such as one or more of intratumoral type 1 conventional dendritic cells (cDC1), macrophages, neutrophils, and circulating monocytes.

[0543] In some respects, anti-TIGIT antagonist antibodies can interact with FcγR of one or more bone marrow cell types (e.g., one or more of cDC1, macrophages, neutrophils, and circulating monocytes) and can induce the mobilization of CD8+ T cells in the blood and / or the expansion of proliferating CD8+ T cells in the tumor bed.

[0544] In some aspects, anti-TIGIT antagonist antibodies are capable of interacting with the MYC-targeting pathway (e.g., upregulating the MYC-targeting pathway). In some aspects, the invention includes detecting levels (e.g., gene expression levels, such as protein or nucleic acid levels) of one or more members of the MYC-targeting pathway (e.g., MYC) in samples from an individual, for example, including detecting levels of one or more members of the MYC-targeting pathway in samples from an individual at time points during or after administration of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and comparing the detected levels with reference expression levels, for example, baseline expression levels in samples from the individual at time points prior to the initiation of treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0545] In some respects, anti-TIGIT antagonist antibodies are IgG1 antibodies, such as tisrelimumab, velbolimab, atelimumab, BGB-A1217, SGN-TGT, EOS084448 (EOS-448), TJ-T6, or AB308. Tirelimumab is an anti-TIGIT monoclonal antibody (mAb) with IgG1 / κFc.

[0546] In other respects, anti-TIGIT antagonist antibodies are IgG4 class antibodies.

[0547] Anti-TIGIT antagonist antibodies (e.g., tirelinumab) that can be used in this invention include compositions containing such antibodies that can bind to PD-1 axis antagonists (e.g., PD-L1 binding antagonists (e.g., anti-PD-L1 antagonist antibodies, such as atezolizumab), PD-1 binding antagonists (e.g., anti-PD-1 antagonist antibodies, such as pembrolizumab) and PD-L2 binding antagonists (e.g., anti-PD-L2 antagonist antibodies)).

[0548] In some embodiments, anti-TIGIT antagonist antibodies inhibit TIGIT signaling. In some embodiments, anti-TIGIT antagonist antibodies inhibit the binding of TIGIT to its binding partner. Exemplary TIGIT binding partners include CD155 (PVR), CD112 (PVRL2 or Nectin-2), and CD113 (PVRL3 or Nectin-3). In some embodiments, anti-TIGIT antagonist antibodies can inhibit the binding between TIGIT and CD155. In some embodiments, anti-TIGIT antagonist antibodies can inhibit the binding between TIGIT and CD112. In some embodiments, anti-TIGIT antagonist antibodies inhibit the binding between TIGIT and CD113. In some embodiments, anti-TIGIT antagonist antibodies inhibit TIGIT-mediated cell signaling in immune cells. In some embodiments, anti-TIGIT antagonist antibodies inhibit TIGIT by consuming regulatory T cells (e.g., when binding to FcγR).

[0549] In some embodiments, the anti-TIGIT antibody is a monoclonal antibody. In some embodiments, the anti-TIGIT antibody is an antibody fragment selected from the group consisting of Fab, Fab'-SH, Fv, scFv, and (Fab')2 fragments. In some embodiments, the anti-TIGIT antibody is a humanized antibody. In some embodiments, the anti-TIGIT antibody is a human antibody. In some embodiments, the anti-TIGIT antibody described herein binds to human TIGIT. In some embodiments, the anti-TIGIT antibody is an Fc fusion protein.

[0550] In some embodiments, the anti-TIGIT antibody is selected from the group consisting of: tirelinumab (MTIG7192A, RG6058 or RO7092284), vimbrolizumab (MK-7684), EOS884448 (EOS-448), SEA-TGT (SGN-TGT)), BGB-A1217, IBI939, M6223, AB308, AB154, TJ-T6, MG1131, NB6253, HLX301, HLX53, SL-9258 (TIGIT-Fc-LIGHT), STW264 and YBL-012. In some embodiments, the anti-TIGIT antibody is selected from the group consisting of: tirelinumab (MTIG7192A, RG6058, or RO7092284), vimbrolizumab (MK-7684), EOS-448, and SEA-TGT (SGN-TGT). The anti-TIGIT antibody may be tirelinumab (MTIG7192A, RG6058, or RO7092284).

[0551] Non-limiting examples of anti-TIGIT antibodies that can be used with the methods disclosed herein and methods for their preparation are disclosed in PCT publications WO2018183889A1, WO2019129261A1, WO2016106302A9, WO2018033798A1, WO2020020281A1, WO2019023504A1, WO2017152088A1, WO2016028656A1, WO2017030823A2, WO2018204405A1, WO2019152574A1 and WO2020041541A2; and US Patents US10,189,902, US10,213,505, and US10,213,505. The patents described in US 10,124,061, 10,537,633, and 10,618,958; and US U.S. Patent Publications 2020 / 0095324, 2019 / 0112375, 2018 / 0371083, and 2020 / 0062859, each of which is incorporated herein by reference in its entirety.Other non-limiting examples of anti-TIGIT antibodies that can be used with the methods disclosed herein and their preparation methods are disclosed in PCT publications WO2018204363A1, WO2018047139A1, WO2019175799A2, WO2018022946A1, WO2015143343A2, WO2018218056A1, WO2019232484A1, WO2019079777A1, WO2018128939A1, WO2017196867A1, WO2019154415A1, WO2019062832A1, WO2018234793A3, WO2018102536A1, and WO2019137548A1. 1. WO2019129221A1, WO2018102746A1, WO2018160704A9, WO2020041541A2, WO2019094637A9, WO2017037707A1, WO2019168382A1, WO2006124667A3, WO2017021526A1, WO2017184619A2, WO2017048824A1, WO2019032619A9, WO2018157162A1, WO2020176718A1, WO2020047329A1, WO2020047329A1, WO2018220446A9; US Patent No. US 9,617,338, US 9,567,399, US 10,604,576 and US 9,994,637; and US Patent Publication No. US2018 / 0355040, US 2019 / 0175654、US2019 / 0040154、US2019 / 0382477、US2019 / 0010246、US2020 / 0164071、US2020 / 0131267、US2019 / 0338032、US 2019 / 0330351, US2019 / 0202917, US2019 / 0284269, US2018 / 0155422, US2020 / 0040082, US2019 / 0263909, US2018 / 0185480, US The descriptions in 2019 / 0375843, US2017 / 0037133, US2019 / 0077869, US2019 / 0367579, US2020 / 0222503, US2020 / 0283496, CN109734806A and CN110818795A are each incorporated herein by reference in their entirety.

[0552] Anti-TIGIT antibodies that can be used in the methods disclosed herein include BGB-A1217, M6223, IBI939, EOS-448, vimbrolizumab (MK-7684), and SEA-TGT (SGN-TGT). Other anti-TIGIT antibodies that can be used with the methods disclosed herein include AAGEN1307; AAGEN1777; antibody clones pab2197 and pab2196 (Agenus Inc.); antibody clones TBB8, TDC8, 3TB3, 5TB10 and D1Y1A (Anhui Anke Biotechnology Group Co., Ltd.); antibody clones MAB1, MAB2, MAB3, MAB4, MAB5, MAB6, MAB7, MAB8, MAB9, MAB10, MAB11, MAB12, MAB13, MAB14, MAB15, MAB16, MAB17, MAB18, MAB19, MAB20, and MAB21 (Astellas Pharmaceuticals). Pharma / Potenza Therapeutics), antibody clones hu1217-1-1 and hu1217-2-2 (BeiGene), antibody clones 4D4 and 19G (Brigham & Women's Hospital), antibody clones 11G11, 10D7, 15A6, 22G2, TIGIT G2a, and TIGIT G1 D265A, including such antibodies with modified heavy chain constant regions (Bristol-Myers Squibb). Squibb); antibody clones 10A7, CPA.9.086, CPA.9.083.H4 (S241P), CPA.9.086.H4 (S241P), CHA.9.547.7.H4 (S241P) and CHA.9.547.13.H4 (S241P) (Compugen); anti-PVRIG / anti-TIGIT bispecific antibody (Compugen), antibody clones 315293, 328189, 350426, 326504 and 331672 (Fred Hutchinson Cancer Research Center); antibody clones T-01, T-02, T-03, T-04, T-05, T-06, T-07, T-08, T-09 and T-10 (Gensun BioPharma Inc.);Antibody clones 1H6, 2B11, 3A10, 4A5, 4A9, 4H5, 6A2, 6B7, 7F4, 8E1, 8G3, 9F4, 9G6, 10C1, 10F10, 11G4, 12B7, 12C8, 15E9, 16C11, 16D6, and 16E10 (Hefei Ruida Immunotherapy Research Institute Co., Ltd.); Antibody clones h3C5H1, h3C5H2, h3C5H3, h3C5H4, h3C5H3-1, h3C5H3-2, h3C5H3-3, h3C5L1, and h3C5L2 (IGM Biosciences). Inc.); antibody clones 90D9, 101E1, 116H8, 118A12, 131A12, 143B6, 167F7, 221F11, 222H4, 327C9, 342A9, 344F2, 349H6 and 350D10 (I-Mab Biopharma); antibody clones ADI-27238, ADI-30263, ADI-30267, ADI-30268, ADI-27243, ADI-30302, ADI-30336, ADI-272 78. ADI-30193, ADI-30296, ADI-27291, ADI-30283, ADI-30286, ADI-30288, ADI27297, ADI-30272, ADI-30278, AD I-27301, ADI-30306, and ADI-30311 (Innovent Biologics, Inc.); antibody clones 26518, 29478, 26452, 29487, 29489, 31282, 26486, 29494, 29499, 26521, 29513, 26493, 29520, 29523, 29527, 31288, 32919, 32931, 26432, and 32959 (iTeos) Therapeutics); antibody clones m1707, m1708, m1709, m1710, m1711, h1707, h1708, h1709, h1710 and h1711 (Jiangsu Hengrui Medicine Co., Ltd.); antibody clones TIG1, TIG2 and TIG3 (JNBiosciences LLC); antibody clones (e.g., KY01, KY02, KY03, KY04, KY05, KY06, KY07, KY08, KY09, KY10, K11, K12, K13, K14, K15, K16, K17, K18, K19, K20, K21, K22, K23), Kymab TIGIT (antibody 2) and Tool TIGIT (antibody 4) (Kymab Limited);Bispecific antibodies 1D05 / internal anti-TIGIT (Bispecific 1) containing the 1D05 (anti-PD-L1) natural variable domain and the Kymab TIGIT antigen-binding site (ABS) domain; internal anti-TIGIT / 1D05 (Bispecific 2) containing the Kymab TIGIT natural variable domain and the 1D05 ABS domain; Tool anti-TIGIT / Tool anti-PD-L1 (Bispecific 3) containing the Toon anti-TIGIT natural variable domain and the Tool anti-PD-L1 ABS domain; and Tool anti-PD-L1 / Tool anti-TIGIT (Bispecific 4) containing the Tool anti-PD-L1 natural variable domain and the Tool anti-TIGIT ABS domain (Kymab TIGIT / Tool anti-TIGIT / Tool anti-TIGIT / Tool anti-TIGIT / Tool anti-TIGIT / Tool anti-TIGIT / Tool anti-TIGIT (Bispecific 4) containing the Kymab TIGIT natural variable domain and the Tool anti-TIGIT ABS domain. Limited); antibody clones and clone variants 14D7, 26B10, Hu14D7, Hu26B10, 14A6, Hu14A6, 28H5, 31C6, Hu31C6, 25G10, MBS43, 37D10, 18G10, 11A11, c18G10 and LB155.14A6.G2.A8 (Merck); etelidomide Anti-(OMP-313M32) (MereoBioPharma); Antibody clones 64G1E9B4, 100C4E7D11, 83G5H11C12, 92E9D4B4, 104G12E12G2, 121C2F10B5, 128E3F10F3F2, 70A11A8E6, 11D8E124A, 16F10H12C11, 8 F2D8E7, 48B5G4E12, 139E2C2D2, 128E3G7F5, AS19584, AS19852, AS19858, AS19886, AS1 9887, AS19888, AS20160, AS19584VH26, AS19584VH29, AS19584VH30, AS19584VH31, AS1 9886VH5, AS19886VH8, AS19886VH9, AS19886VH10, AS19886VH19, AS19886VH20, AS19584VH28-Fc, AS19886VH5-Fc, AS19886VH8-Fc, AS19584-Fc and AS19886-Fc (Nanjing Legend Biotechnology Co., Ltd.);Antibody clones ARE clones: Ab58, Ab69, Ab75, Ab133, Ab177, Ab122, Ab86, Ab180, Ab83, Ab26, Ab20, Ab147, Ab12, Ab66, Ab176, Ab96, Ab123, Ab109, Ab149, Ab34, Ab61, Ab64, Ab105, Ab108, Ab178, Ab166, Ab2 9. Ab135, Ab171, Ab194, Ab184, Ab164, Ab183, Ab158, Ab55, Ab136, Ab39, Ab159, Ab151, Ab139, A b107, Ab36, Ab193, Ab115, Ab106, Ab13f8, Ab127, Ab165, Ab155, Ab19, Ab6, Ab187, Ab179, Ab65, A b114, Ab102, Ab94, Ab163, Ab110, Ab80, Ab92, Ab117, Ab162, Ab121, Ab195, Ab84, Ab161, Ab198, Ab24, Ab98, Ab116, Ab174, Ab196, Ab51, Ab91, Ab185, Ab23, Ab7, Ab95, Ab100, Ab140, Ab145, Ab1 50, Ab168, Ab54, Ab77, Ab43, Ab160, Ab82, Ab189, Ab17, Ab103, Ab18, Ab130, Ab132, Ab134, Ab144; ARG clones: Ab2, Ab47, Ab49, Ab31, Ab53, Ab40, Ab5, Ab9, Ab48, Ab4, Ab10, Ab37, Ab33, Ab42, Ab45;ARV clones: Ab44, Ab97, Ab81, Ab188, Ab186, Ab62, Ab57, Ab192, Ab73, Ab60, Ab28, Ab32, Ab78, Ab14, Ab152, Ab72, Ab137, Ab128, Ab169, Ab87, Ab74, Ab 172, Ab153, Ab120, Ab13, Ab113, Ab16, Ab56, Ab129, Ab50, Ab90, Ab99, Ab3, Ab148, Ab124, Ab22, Ab41, Ab119, Ab157, Ab27, Ab15, Ab191, Ab190, Ab79 Ab181, Ab146, Ab167, Ab88, Ab199, Ab71, Ab85, Ab59, Ab141, Ab68, Ab143, Ab46, Ab197, Ab175, Ab156, Ab63, Ab11, Ab182, Ab89, Ab8, Ab101, Ab25, Ab154, Ab21, Ab111, Ab118, Ab173, Ab38, Ab76, Ab131, Ab1, Ab67, Ab70, Ab170, Ab30, Ab93, Ab142, Ab104, Ab112, Ab35, Ab126, and Ab125 (Rigel Pharmaceuticals). Pharmaceuticals, Inc.); CASC-674 (Seattle Genetics); Antibody clones 2, 2C, 3, 5, 13, 13A, 13B, 13C, 13D, 14, 16, 16C, 16D, 16E, 18, 21, 22, 25, 25A, 25B, 25C, 25D, 25E, 27, 54, 13IgG2a (fucosylated), 13hIgG1 (wild-type), and 13LALA-PG (Seattle Genetics); JS006 (Shanghai Junshi Biosciences Co., Ltd.); Anti-TIGIT Fc antibody and bispecific antibody PD1 xTIGIT (Xencor), antibody clones VSIG9#1 (Vsig9.01) and 258-CS1#4 (#4) (Yissum Research Development Company of The Hebrew University of Jerusalem). Ltd.)); YH29143 (Yuhan Co,Ltd.);Antibody clones S02, S03, S04, S05, S06, S11, S12, S14, S19, S32, S39, S43, S62, S64, F01, F02, F03, F04, 32D7, 101H3, 10A7, and 1F4 (Yuhan Co., Ltd.); anti-zB7R1 clones 318.4.1.1 (E9310), 318.28.2.1 (E9296), 318.39.1.1 (E9311), 318.59.3.1 (E9400), and 318.77.1.10 (ZymoGenetics, Inc.).

[0553] In some embodiments, the anti-TIGIT antibody is selected from the group consisting of tirelinumab, BGB-A1217, M6223, IBI939, EOS884448 (EOS-448), vimbrolizumab (MK-7684), and SEA-TGT (SGN-TGT). ASP874 (PTZ-201) is the anti-TIGIT monoclonal antibody described in PCT Publication No. WO2018183889A1 and US Patent Publication No. 2020 / 0095324. BGB-A1217 is the anti-TIGIT antibody as described in PCT Publication No. WO2019129261A1. IBI939 is the anti-TIGIT antibody as described in PCT Publication No. WO2020020281A1. EOS884448 (EOS-448) is the anti-TIGIT antibody described in PCT Publication No. WO2019023504A1. Vimbalimab (MK-7684) is the anti-TIGIT antibody described in PCT Publications Nos. WO2016028656A1, WO2017030823A2, WO2018204405A1 and / or WO2019152574A1, US Patent No. 10,618,958 and US Patent Publication No. 2018 / 0371083. SEA-TGT (SGN-TGT) is the anti-TIGIT antibody as described in PCT Publication No. WO2020041541A2 and US Patent Publication No. 2020 / 0062859.

[0554] In some embodiments, the anti-TIGIT antagonist antibody is tirelimumab (CAS Registry No.: 1918185-84-8). Tirelimumab (Genentech) is also known as MTIG7192A, RG6058, or RO7092284. Tirelimumab is also known by PCT publication numbers WO2003072305A8, WO2004024068A3, WO2004024072A3, WO2009126688A2, WO2015009856A2, WO2016011264A1, WO2016109546A2, WO2017053748A2, and WO2019165434A1, as well as US patent publication numbers 2017 / 0044256 and 2017 / 00371. 27. The anti-TIGIT antagonist monoclonal antibodies described in US patent numbers US9873740B2, US10626174B2, US10611836B2, US9499596B2, US8431350B2, US10047158B2 and US10017572B2.

[0555] In some embodiments, an anti-TIGIT antibody comprises at least one, two, three, four, five, or six complementarity-determining regions (CDRs) of any anti-TIGIT antibody disclosed herein. In some embodiments, an anti-TIGIT antibody comprises six CDRs of any anti-TIGIT antibody disclosed herein. In some embodiments, an anti-TIGIT antibody comprises six CDRs of any antibody selected from the group consisting of tirelinumab, BGB-A1217, M6223, IBI939, EOS884448 (EOS-448), vimbrolizumab (MK-7684), and SEA-TGT (SGN-TGT).

[0556] In some embodiments, the anti-TIGIT antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the heavy chain variable region (VH) sequence of any of the anti-TIGIT antibodies disclosed herein, and the light chain comprises the light chain variable region (VL) of the same antibody. In some embodiments, the anti-TIGIT antibody comprises the VH and VL of an anti-TIGIT antibody selected from the group consisting of tirelinumab, BGB-A1217, M6223, IBI939, EOS884448 (EOS-448), vimbrolizumab (MK-7684), and SEA-TGT (SGN-TGT).

[0557] In some embodiments, the anti-TIGIT antibody comprises the heavy and light chains of any anti-TIGIT antibody disclosed herein. In some embodiments, the anti-TIGIT antibody comprises the heavy and light chains of an anti-TIGIT antibody selected from the group consisting of tirelenumab, BGB-A1217, M6223, IBI939, EOS884448 (EOS-448), vimbrolizumab (MK-7684), and SEA-TGT (SGN-TGT).

[0558] In some embodiments, the anti-TIGIT antagonist antibody according to any of the embodiments described herein may combine, alone or in combination, any of the features described in Section IV(C) below.

[0559] B. PD-1 axis binding antagonists

[0560] This article provides a method for treating cancer in a subject (e.g., a human), the method comprising administering to the subject an effective amount of a PD-1 axis binding antagonist. PD-1 axis binding antagonists may include PD-L1 binding antagonists, PD-1 binding antagonists, and PD-L2 binding antagonists. Any suitable PD-1 axis binding antagonist may be used.

[0561] 1. PD-L1 binding antagonists

[0562] In some cases, PD-L1 binding antagonists inhibit the binding of PD-L1 to one or more of its ligand-binding partners. In other cases, PD-L1 binding antagonists inhibit the binding of PD-L1 to PD-1. In still other cases, PD-L1 binding antagonists inhibit the binding of PD-L1 to B7-1. In some cases, PD-L1 binding antagonists inhibit the binding of PD-L1 to both PD-1 and B7-1. PD-L1 binding antagonists can be, but are not limited to, antibodies, their antigen-binding fragments, immunoadhesins, fusion proteins, oligopeptides, or small molecules. In some cases, PD-L1 binding antagonists are small molecules that inhibit PD-L1 (e.g., GS-4224, INCB086550, MAX-10181, INCB090244, CA-170, or ABSK041). In some cases, PD-L1 binding antagonists are small molecules that inhibit both PD-L1 and VISTA. In some cases, the PD-L1 binding antagonist is CA-170 ...

Claims

1. A method for identifying an individual with cancer who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, the method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a tumor-associated macrophage (TAM) signature score therefrom, wherein a TAM signature score higher than a reference TAM signature score identifies the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

2. A method for selecting a therapy for an individual having cancer, the method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a TAM signature score therefrom, wherein a TAM signature score higher than a reference TAM signature score identifies the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

3. The method of claim 1 or 2, wherein the individual has a TAM signature score in the sample that is higher than a reference TAM signature score, and the method further comprises administering to the individual an effective amount of a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

4. A method of treating an individual suffering from cancer, the method comprising: (a) detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a TAM signature score therefrom, wherein the TAM signature score is higher than a reference TAM signature score and thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody; and (b) administering to the individual an effective amount of a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

5. A method for treating an individual having cancer, the method comprising administering a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody to the individual, wherein the individual has been determined to have a TAM signature score that is higher than a reference TAM signature score, thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, and wherein the TAM signature score is based on the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO detected in a sample from the individual.

6. The method of any one of claims 1 to 5, wherein the sample is obtained from the individual prior to treatment with the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody.

7. The method according to any one of claims 1 to 6, wherein the benefit is an increase in progression-free survival (PFS), objective response rate (ORR), or overall survival (OS).

8. The method according to any one of claims 1 to 7, wherein the reference TAM signature score is a pre-specified TAM signature score.

9. The method according to any one of claims 1 to 8, wherein the reference TAM signature score is a TAM signature score in a reference population.

10. The method of claim 9, wherein the TAM signature score in the reference population is the median TAM signature score of the reference population.

11. The method of claim 9 or 10, wherein the reference population is a population of individuals suffering from the cancer.

12. The method according to any one of claims 1 to 11, wherein the TAM signature score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in the sample from the individual.

13. The method of claim 12, wherein the TAM signature score is the average of the normalized expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual.

14. The method according to any one of claims 1 to 4 and 6 to 13, wherein the method comprises further detecting the expression level of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD in the sample from the individual.

15. The method of claim 14, wherein the TAM signature score is the average of the expression levels of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, and ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in the sample from the individual.

16. The method of claim 14 or 15, wherein the method comprises further detecting the expression level of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in the sample from the individual and determining the TAM signature score therefrom, wherein the TAM signature score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in the sample from the individual.

17. The method of claim 5, wherein the expression level of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD has been detected in the sample from the individual.

18. The method of claim 17, wherein the TAM signature score is the average of the expression levels of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, and ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in the sample from the individual.

19. The method of claim 17 or 18, wherein the expression level of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD has been detected in the sample from the individual and the TAM signature score has been determined therefrom, wherein the TAM signature score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD in the sample from the individual.

20. A method for monitoring the response of an individual with cancer to a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, the method comprising detecting MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, An expression level of one or more of LCAT and LIRA3, wherein an increase in the expression level of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT and LIRA3 relative to their respective reference expression levels predicts an individual who may respond to the treatment comprising the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody.

21. The method of claim 20, wherein the expression level of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT and LIRA3 is detected three weeks after starting the treatment comprising the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody.

22. The method of claim 20 or 21, wherein the expression level of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 is detected six weeks after starting the treatment comprising the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody.

23. The method of any one of claims 20 to 22, wherein the expression level of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT and LIRA3 is increased relative to the respective reference expression levels, thereby predicting that the individual may respond to the treatment comprising the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody, and the method further comprises administering an additional dose of the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody to the individual.

24. The method of any one of claims 20 to 23, wherein the response to treatment is an increase in PFS or OS.

25. The method of any one of claims 20 to 24, wherein the reference expression level is a baseline expression level of a sample from the individual at a time point prior to starting the treatment comprising the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody.

26. A method for identifying an individual with cancer who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, the method comprising detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a regulatory T cell (Treg) signature score therefrom, wherein a Treg signature score higher than a reference Treg signature score identifies the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

27. A method for selecting a therapy for an individual having cancer, the method comprising detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a Treg signature score therefrom, wherein a Treg signature score higher than a reference Treg signature score identifies the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

28. The method of claim 26 or 27, wherein the individual has a Treg characteristic score in the sample that is higher than a reference Treg characteristic score, and the method further comprises administering to the individual an effective amount of a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

29. A method of treating an individual suffering from cancer, the method comprising: (a) detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a Treg signature score therefrom, wherein the Treg signature score is higher than a reference Treg signature score and thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody; and (b) administering to the individual an effective amount of a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

30. A method for treating an individual having cancer, the method comprising administering a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody to the individual, wherein the individual has been determined to have a Treg signature score that is higher than a reference Treg signature score, thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, and wherein the Treg signature score is based on the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in a sample from the individual.

31. The method of any one of claims 26 to 30, wherein the sample is obtained from the individual prior to treatment with a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

32. The method of any one of claims 26 to 31, wherein the benefit is an increase in PFS, ORR, or OS.

33. The method of any one of claims 26 to 32, wherein the reference Treg signature score is a pre-specified Treg signature score.

34. The method of any one of claims 26 to 33, wherein the reference Treg signature score is a Treg signature score in a reference population.

35. The method of claim 34, wherein the Treg signature score in the reference population is the median Treg signature score of the reference population.

36. The method of claim 34 or 35, wherein the reference population is a population of individuals suffering from the cancer.

37. The method of any one of claims 26 to 36, wherein the Treg signature score is the average of the expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in the sample from the individual.

38. The method of claim 37, wherein the Treg signature score is the average of normalized expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in the sample from the individual.

39. The method according to any one of claims 1 to 38, wherein the expression level is a nucleic acid expression level or a protein expression level.

40. The method of claim 39, wherein the expression level is a nucleic acid expression level.

41. The method of claim 40, 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.

42. The method of claim 40 or 41, wherein the nucleic acid expression level is mRNA expression level.

43. The method of claim 42, wherein the mRNA expression level is determined by RNA-seq.

44. The method of claim 39, wherein the expression level is a protein expression level.

45. The method of claim 44, wherein the protein expression level is determined by mass spectrometry.

46. ​​The method of any one of claims 1 to 19 and 26 to 38, wherein the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

47. The method of claim 46, wherein the sample is a tissue sample.

48. The method of claim 47, wherein the tissue sample is a tumor tissue sample.

49. The method of claim 48, wherein the tumor tissue sample is a biopsy.

50. The method of any one of claims 20 to 25, wherein the sample is a serum sample.

51. The method of any one of claims 46 to 50, wherein the sample is an archived sample, a fresh sample, or a frozen sample.

52. The method of any one of claims 1 to 51, wherein the sample has been determined to have a PD-L1 positive tumor cell fraction by immunohistochemistry (IHC) assay.

53. The method of claim 52, wherein the PD-L1 positive tumor cell fraction is determined by positive staining with an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody is SP263, 22C3, SP142 or 28-8.

54. The method of claim 53, wherein the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with the anti-PD-L1 antibody SP263.

55. The method of claim 54, wherein the PD-L1 positive tumor cell fraction is calculated using the Ventana SP263 IHC assay.

56. The method of claim 53, wherein the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with the anti-PD-L1 antibody 22C3.

57. The method of claim 56, wherein the PD-L1 positive tumor cell fraction is calculated using the pharmDx 22C3 IHC assay.

58. The method of any one of claims 1 to 57, wherein the cancer is lung cancer.

59. The method of claim 58, wherein the lung cancer is non-small cell lung cancer (NSCLC).

60. The method of any one of claims 1 to 59, wherein the anti-TIGIT antagonist antibody comprises the following hypervariable regions (HVRs): (a) HVR-H1 comprising the amino acid sequence of SNSAAWN (SEQ ID NO: 1); (b) HVR-H2 comprising the amino acid sequence of KTYYRFKWYSDYAVSVKG (SEQ ID NO: 2); (c) HVR-H3 comprising the amino acid sequence of ESTTYDLLAGPFDY (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KSSQTVLYSSNNKKYLA (SEQ ID NO: 4); (e) HVR-L2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 5); and (f) HVR-L3 comprising the amino acid sequence of QQYYSTPFT (SEQ ID NO: 6).

61. The method of claim 60, wherein the anti-TIGIT antagonist antibody further comprises the following light chain variable region FR: (a) FR-L1 comprising the amino acid sequence of DIVMTQSPDSLAVSLGERATINC (SEQ ID NO: 7); (b) FR-L2 comprising the amino acid sequence of WYQQKPGQPPNLLIY (SEQ ID NO: 8); (c) FR-L3 comprising the amino acid sequence of GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 9); and (d) FR-L4 comprising the amino acid sequence of FGPGTKVEIK (SEQ ID NO: 10).

62. The method of claim 60 or 61, wherein the anti-TIGIT antagonist antibody further comprises the following heavy chain variable region FR: (a) FR-H1, comprising an amino acid sequence of X1VQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 11), wherein X1 is Q or E; (b) FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); (c) FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and (d) FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14).

63. The method of claim 62, wherein X1 is Q.

64. The method of claim 62, wherein X1 is E.

65. The method of any one of claims 60 to 64, wherein the anti-TIGIT antagonist antibody comprises: (a) a VH domain comprising EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSL QLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS(SEQ ID NO:17) or QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSL an amino acid sequence having at least 95% sequence identity to the amino acid sequence of QLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO: 18); (b) a VL domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAWYQQKPGQPPNLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPFTFGPGTKVEIK (SEQ ID NO: 19); or (c) VH domain as in (a) and VL domain as in (b).

66. The method of claim 65, wherein the anti-TIGIT antagonist antibody comprises: (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 17 or 18; and (b) A VL domain comprising the amino acid sequence of SEQ ID NO:

19.

67. The method of claim 66, wherein the anti-TIGIT antagonist antibody comprises: (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 17; and (b) A VL domain comprising the amino acid sequence of SEQ ID NO:

19.

68. The method of any one of claims 1 to 62 and 64 to 67, wherein the anti-TIGIT antagonist antibody comprises: (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 33; and (b) a light chain comprising the amino acid sequence of SEQ ID NO:

34.

69. The method of any one of claims 1 to 68, wherein the anti-TIGIT antagonist antibody is a monoclonal antibody.

70. The method of any one of claims 1 to 69, wherein the anti-TIGIT antagonist antibody is a human antibody.

71. The method of any one of claims 1 to 70, wherein the anti-TIGIT antagonist antibody is a full-length antibody.

72. The method of any one of claims 1 to 71, wherein the anti-TIGIT antagonist antibody exhibits effector function.

73. The method of any one of claims 1 to 72, wherein the anti-TIGIT antagonist antibody comprises an Fc domain capable of interacting with an Fcγ receptor (FcγR).

74. The method of any one of claims 1 to 73, wherein the anti-TIGIT antagonist antibody is an IgG class antibody.

75. The method of claim 74, wherein the IgG class antibody is an IgG1 subclass antibody.

76. The method of any one of claims 1 to 62 and 64 to 75, wherein the anti-TIGIT antagonist antibody is tirilimumab.

77. The method of any one of claims 1 to 67, 69 and 70, wherein the anti-TIGIT antagonist antibody is a TIGIT-binding antibody fragment selected from the group consisting of Fab, Fab', Fab'-SH, Fv, single-chain variable fragment (scFv) and (Fab')2 fragment.

78. The method of any one of claims 1 to 59, wherein the anti-TIGIT antagonist antibody is vibriostat, etilizumab, EOS084448, SGN-TGT, TJ-T6, BGB-A1217, or AB308.

79. The method of any one of claims 1 to 78, wherein the PD-1 axis binding antagonist is selected from the group consisting of a PD-L1 binding antagonist, a PD-1 binding antagonist, and a PD-L2 binding antagonist.

80. The method of claim 79, wherein the PD-1 axis binding antagonist is a PD-L1 binding antagonist.

81. The method of claim 80, wherein the PD-L1 binding antagonist inhibits binding of PD-L1 to one or more of its ligand binding partners.

82. The method of claim 81, wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1, B7-1, or to both PD-1 and B7-1.

83. The method of any one of claims 80 to 82, wherein the PD-L1 binding antagonist is an anti-PD-L1 antagonist antibody.

84. The method of claim 83, wherein the anti-PD-L1 antagonist antibody is atezolizumab, MDX-1105, durvalumab, avelumab, SHR-1316, CS1001, envolimab, TQB2450, ZKAB001, LP-002, CX-072, IMC-001, KL-A167, APL-502, coxilimab, lodalimab, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, RC98, PDL-GEX, KD036, KY1003, YBL-007, or HS-636.

85. The method of claim 84, wherein the anti-PD-L1 antagonist antibody is atezolizumab.

86. The method of claim 83, wherein the anti-PD-L1 antagonist antibody comprises the following HVRs: (a) an HVR-H1 sequence comprising the amino acid sequence of GFTFSDSWIH (SEQ ID NO: 20); (b) an HVR-H2 sequence comprising AWISPYGGSTYYADSVKG (SEQ ID NO: 21) amino acid sequence; (c) an HVR-H3 sequence comprising the amino acid sequence of RHWPGGFDY (SEQ ID NO: 22); (d) an HVR-L1 sequence comprising the amino acid sequence of RASQDVSTAVA (SEQ ID NO: 23); (e) an HVR-L2 sequence comprising the amino acid sequence of SASFLYS (SEQ ID NO: 24); and (f) an HVR-L3 sequence comprising the amino acid sequence of QQYLYHPAT (SEQ ID NO: 25).

87. The method of claim 86, wherein the anti-PD-L1 antagonist antibody comprises: (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 26; (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 27; or (c) VH domain as in (a) and VL domain as in (b).

88. The method of claim 85, wherein the anti-PD-L1 antagonist antibody comprises: (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 26; and (b) a VL domain comprising the amino acid sequence of SEQ ID NO:

27.

89. The method of claim 88, wherein the anti-PD-L1 antagonist antibody comprises: (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 28; and (b) a light chain comprising the amino acid sequence of SEQ ID NO:

29.

90. The method of any one of claims 86 to 89, wherein the anti-PD-L1 antagonist antibody is a monoclonal antibody.

91. The method of any one of claims 86 to 90, wherein the anti-PD-L1 antagonist antibody is a humanized antibody.

92. The method of any one of claims 86 to 91, wherein the anti-PD-L1 antagonist antibody is a full-length antibody.

93. The method of any one of claims 86 to 88, 90 and 91, wherein the anti-PD-L1 antagonist antibody is a PD-L1 binding antibody fragment selected from the group consisting of Fab, Fab', Fab'-SH, Fv, scFv and (Fab')2 fragments.

94. The method of any one of claims 86 to 92, wherein the anti-PD-L1 antagonist antibody is an IgG class antibody.

95. The method of claim 94, wherein the IgG class antibody is an IgG1 subclass antibody.

96. The method of claim 79, wherein the PD-1 axis binding antagonist is a PD-1 binding antagonist.

97. The method of claim 96, wherein the PD-1 binding antagonist inhibits binding of PD-1 to one or more of its ligand binding partners.

98. The method of claim 97, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1, PD-L2, or to both PD-L1 and PD-L2.

99. The method of any one of claims 96 to 98, wherein the PD-1 binding antagonist is an anti-PD-1 antagonist antibody.

100. The method of claim 99, wherein the anti-PD-1 antagonist antibody is nivolumab, pembrolizumab, MEDI-0680, spartalizumab, cemiplizumab, BGB-108, palolizumab, carrelizumab, sintilimab, tislelizumab, toripalimab, dotalimumab, rivulimab, sazanlimab, penampalimab, CS1003, HLX10, SCT-I10A, sepalizumab, batilizumab, genoluzumab, BI 754091, silimumab, YBL-006, BAT1306, HX008, brigalimumab, AMG 404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, AM0001, ENUM 244C8, ENUM 388D4, STI-1110, AK-103, or hAb21.

101. The method of any one of claims 79 and 96 to 98, wherein the PD-1 binding antagonist is an Fc fusion protein.

102. The method of claim 101, wherein the Fc fusion protein is AMP-224.

103. The method of any one of claims 1 to 102, wherein the individual is a human.

104. Use of a PD-1 axis binding antagonist and / or an anti-TIGIT antagonist antibody in the manufacture of a medicament for treating an individual having cancer, wherein the individual has been determined to have a TAM signature score that is higher than a reference TAM signature score, thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, and wherein the TAM signature score is based on the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO detected in a sample from the individual.

105. The use according to claim 104, wherein the sample is obtained from the individual prior to treatment with the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody.

106. The use according to claim 104 or 105, wherein the benefit is an increase in progression-free survival (PFS), objective response rate (ORR) or overall survival (OS).

107. The use according to any one of claims 104 to 106, wherein the reference TAM signature score is a pre-specified TAM signature score.

108. The use according to any one of claims 104 to 107, wherein the reference TAM signature score is the TAM signature score in a reference population.

109. The use according to claim 108, wherein the TAM signature score in the reference population is the median TAM signature score of the reference population.

110. The use according to claim 108 or 109, wherein the reference population is a population of individuals suffering from the cancer.

111. The use according to any one of claims 104 to 110, wherein the TAM signature score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in the sample from the individual.

112. The use according to claim 111, wherein the TAM signature score is the average of the normalized expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in the sample from the individual.

113. The use according to claim 104, wherein the expression level of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD has been detected in the sample from the individual.

114. The use according to claim 113, wherein the TAM signature score is the average of the expression levels of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, and ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in the sample from the individual.

115. The use according to claim 113 or 114, wherein the expression level of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD has been detected in the sample from the individual and the TAM signature score has been determined therefrom, wherein the TAM signature score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD in the sample from the individual.

116. Use of a PD-1 axis binding antagonist and / or an anti-TIGIT antagonist antibody in the manufacture of a medicament for treating an individual having cancer, wherein the individual has been determined to have a Treg signature score that is higher than a reference Treg signature score, thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, and wherein the Treg signature score is based on the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in a sample from the individual.

117. The use according to claim 116, wherein the sample is obtained from the individual prior to treatment with a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody.

118. The use according to claim 116 or 117, wherein the benefit is an increase in PFS, ORR or OS.

119. The use according to any one of claims 116 to 118, wherein the reference Treg signature score is a pre-specified Treg signature score.

120. The use according to any one of claims 116 to 119, wherein the reference Treg signature score is the Treg signature score in a reference population.

121. The use according to claim 120, wherein the Treg signature score in the reference population is the median Treg signature score of the reference population.

122. The use according to claim 120 or 121, wherein the reference population is a population of individuals suffering from the cancer.

123. The use according to any one of claims 116 to 122, wherein the Treg signature score is the average of the expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in the sample from the individual.

124. The use according to claim 123, wherein the Treg signature score is the average of the normalized expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in the sample from the individual.

125. The use according to any one of claims 104 to 124, wherein the expression level is a nucleic acid expression level or a protein expression level.

126. The use according to claim 125, wherein the expression level is a nucleic acid expression level.

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 claim 125, wherein the expression level is a protein expression level.

131. The use according to claim 130, wherein the protein expression level is determined by mass spectrometry.

132. The use according to any one of claims 104 to 131, wherein the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

133. The use according to claim 132, wherein the sample is a serum sample.

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

135. The use according to claim 134, wherein the tissue sample is a tumor tissue sample.

136. The use according to claim 135, wherein the tumor tissue sample is a biopsy.

137. The use according to any one of claims 132 to 136, wherein the sample is an archived sample, a fresh sample or a frozen sample.

138. The use according to any one of claims 104 to 137, wherein the sample has been determined to have a PD-L1 positive tumor cell fraction by immunohistochemistry (IHC) assay.

139. The use according to claim 138, wherein the PD-L1 positive tumor cell fraction is determined by positive staining with an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody is SP263, 22C3, SP142 or 28-8.

140. The use according to claim 139, wherein the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with the anti-PD-L1 antibody SP263.

141. The use according to claim 140, wherein the PD-L1 positive tumor cell fraction is calculated using the Ventana SP263 IHC assay.

142. The use according to claim 139, wherein the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with the anti-PD-L1 antibody 22C3.

143. The use according to claim 142, wherein the PD-L1 positive tumor cell fraction is calculated using the pharmDx22C3 IHC assay.

144. The use according to any one of claims 104 to 143, wherein the cancer is lung cancer.

145. The use according to claim 144, wherein the lung cancer is non-small cell lung cancer (NSCLC).

146. A PD-1 axis binding antagonist and / or an anti-TIGIT antagonist antibody for use in the treatment of an individual having cancer, wherein the individual has been determined to have a TAM signature score that is higher than a reference TAM signature score, thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, and wherein the TAM signature score is based on the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO detected in a sample from the individual.

147. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 146, wherein the sample is obtained from the individual prior to treatment with the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody.

148. A PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 146 or 147, wherein the benefit is an increase in progression-free survival (PFS), objective response rate (ORR) or overall survival (OS).

149. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 148, wherein the reference TAM signature score is a pre-specified TAM signature score.

150. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 149, wherein the reference TAM signature score is the TAM signature score in a reference population.

151. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 150, wherein the TAM characteristic score in the reference population is the median TAM characteristic score of the reference population.

152. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 150 or 151, wherein the reference population is a population of individuals suffering from the cancer.

153. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 152, wherein the TAM signature score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in the sample from the individual.

154. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 153, wherein the TAM signature score is the average of the normalized expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in the sample from the individual.

155. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 146, wherein the expression level of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD has been detected in the sample from the individual.

156. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 155, wherein the TAM signature score is the average of the expression levels of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, and ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in the sample from the individual.

157. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 155 or 156, wherein the expression level of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD has been detected in the sample from the individual and the TAM signature score has been determined therefrom, wherein the TAM signature score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, MARCO, ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2 and CTSD in the sample from the individual.

158. A PD-1 axis binding antagonist and / or an anti-TIGIT antagonist antibody for use in the treatment of an individual having cancer, wherein the individual has been determined to have a Treg signature score that is higher than a reference Treg signature score, thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody, and wherein the Treg signature score is based on the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in a sample from the individual.

159. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 158, wherein the sample is obtained from the individual prior to treatment with the PD-1 axis binding antagonist and anti-TIGIT antagonist antibody.

160. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 158 or 159, wherein the benefit is an increase in PFS, ORR or OS.

161. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 158 to 160, wherein the reference Treg characteristic score is a pre-specified Treg characteristic score.

162. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 158 to 161, wherein the reference Treg characteristic score is the Treg characteristic score in a reference population.

163. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 162, wherein the Treg characteristic score in the reference population is the median Treg characteristic score of the reference population.

164. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 162 or 163, wherein the reference population is a population of individuals suffering from the cancer.

165. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 158 to 164, wherein the Treg characteristic score is the average of the expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in the sample from the individual.

166. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 165, wherein the Treg characteristic score is the average of the normalized expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in the sample from the individual.

167. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 166, wherein the expression level is a nucleic acid expression level or a protein expression level.

168. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 167, wherein the expression level is a nucleic acid expression level.

169. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody used according to claim 168, 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.

170. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 168 or 169, wherein the nucleic acid expression level is the mRNA expression level.

171. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 170, wherein the mRNA expression level is determined by RNA-seq.

172. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 167, wherein the expression level is a protein expression level.

173. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 172, wherein the protein expression level is determined by mass spectrometry.

174. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 173, wherein the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample or a combination thereof.

175. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 174, wherein the sample is a tissue sample.

176. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 175, wherein the tissue sample is a tumor tissue sample.

177. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 176, wherein the tumor tissue sample is a biopsy.

178. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 174 to 177, wherein the sample is an archived sample, a fresh sample or a frozen sample.

179. A PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 178, wherein the sample has been determined to have a PD-L1 positive tumor cell fraction by immunohistochemistry (IHC) assay.

180. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 179, wherein the PD-L1 positive tumor cell fraction is determined by positive staining with an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody is SP263, 22C3, SP142 or 28-8.

181. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 180, wherein the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with the anti-PD-L1 antibody SP263.

182. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 181, wherein the PD-L1 positive tumor cell fraction is calculated using the Ventana SP263 IHC assay.

183. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 180, wherein the PD-L1 positive tumor cell fraction is greater than or equal to 50%, as determined by positive staining with the anti-PD-L1 antibody 22C3.

184. A PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 183, wherein the PD-L1 positive tumor cell fraction is calculated using the pharmDx 22C3 IHC assay.

185. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 184, wherein the cancer is lung cancer.

186. The PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to claim 185, wherein the lung cancer is non-small cell lung cancer (NSCLC).

187. A method according to any one of claims 1 to 103, a use according to any one of claims 104 to 145, or a PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 186, wherein the anti-TIGIT antagonist antibody is capable of Fc-dependent activation of bone marrow cells, optionally wherein the bone marrow cells are cells selected from the group consisting of intratumoral type 1 conventional dendritic cells (cDC1), macrophages, neutrophils and circulating monocytes.

188. A method according to any one of claims 1 to 103, a use according to any one of claims 104 to 145, or a PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 186, wherein the anti-TIGIT antagonist antibody is capable of interacting with Fcγ receptors (FcγR) on bone marrow cells and is capable of inducing CD8+T cell mobilization in the blood and / or expansion of proliferating CD8+T cells in the tumor bed.

189. A method for identifying an individual with cancer who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function, the method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a tumor-associated macrophage (TAM) signature score therefrom, wherein a TAM signature score higher than a reference TAM signature score identifies the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function.

190. A method for selecting a therapy for an individual having cancer, the method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a TAM signature score therefrom, wherein a TAM signature score higher than a reference TAM signature score identifies the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function.

191. The method of claim 189 or 190, wherein the individual has a TAM signature score in the sample that is higher than a reference TAM signature score, and the method further comprises administering to the individual an effective amount of a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function.

192. A method of treating an individual suffering from cancer, the method comprising: (a) detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from the individual and determining a TAM signature score therefrom, wherein the TAM signature score is higher than a reference TAM signature score and thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function; and (b) administering to the individual an effective amount of a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

193. A method for treating an individual having cancer, the method comprising administering to the individual a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function, wherein the individual has been determined to have a TAM signature score that is higher than a reference TAM signature score, thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function, and wherein the TAM signature score is based on the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO detected in a sample from the individual.

194. A method for monitoring the response of an individual with cancer to a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function, the method comprising detecting MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, L in a sample from the individual at a time point during or after administration of the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody that exhibits effector function. An expression level of one or more of YAM1, LCAT and LIRA3, wherein an increase in the expression level of one or more of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT and LIRA3 relative to their respective reference expression levels predicts an individual who may respond to the treatment comprising the PD-1 axis binding antagonist and the anti-TIGIT antagonist antibody exhibiting effector function.

195. A method for identifying an individual with cancer who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function, the method comprising detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a regulatory T cell (Treg) signature score therefrom, wherein a Treg signature score that is higher than a reference Treg signature score identifies the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function.

196. A method for selecting a therapy for an individual having cancer, the method comprising detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a Treg signature score therefrom, wherein a Treg signature score higher than a reference Treg signature score identifies the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function.

197. The method of claim 195 or 196, wherein the individual has a Treg characteristic score in the sample that is higher than a reference Treg characteristic score, and the method further comprises administering to the individual an effective amount of a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function.

198. A method of treating an individual suffering from cancer, the method comprising: (a) detecting the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from the individual and determining a Treg signature score therefrom, wherein the Treg signature score is higher than a reference Treg signature score and thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function; and (b) administering to the individual an effective amount of a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

199. A method for treating an individual having cancer, the method comprising administering to the individual a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function, wherein the individual has been determined to have a Treg signature score that is higher than a reference Treg signature score, thereby identifying the individual as an individual who may benefit from a treatment comprising a PD-1 axis binding antagonist and an anti-TIGIT antagonist antibody that exhibits effector function, and wherein the Treg signature score is based on the expression level of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in a sample from the individual.

200. The method of any one of claims 189 to 199, wherein the anti-TIGIT antagonist antibody comprises an Fc domain capable of interacting with an Fcγ receptor (FcγR).

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