A method for determining the efficacy of treating lung cancer, comprising an anti-PD-L1 antagonist and an anti-TIGIT antagonist antibody

Abstract

The present invention relates to a prognostic diagnosis method and a treatment method 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

Technical Field

[0001] Sequence Listing This application includes a Sequence Listing submitted electronically in XML format, which is hereby incorporated by reference in its entirety. The XML copy created on June 1, 2023, is named 50474-290WO4_Sequence_Listing_6_1_23 and is 33,392 bytes in size.

[0002] Provided herein are prognostic 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.

Background Art

[0003] Cancer is characterized by the uncontrolled growth of cell subpopulations. Cancer is the leading cause of death in developed countries and the second most common cause of death in developing countries, with over 14 million new cancer cases diagnosed and over 8 million cancer deaths occurring each year. Therefore, cancer care represents a significant and growing societal burden.

[0004] Programmed cell death-1 / programmed cell death ligand-1 (PD-1 / PD-L1) blockade is effective across a wide range of malignancies. However, not all patients benefit, and a significant proportion of initial responders ultimately relapse. One approach to expand and extend the impact of cancer immunotherapy has been to target additional immune checkpoints. One such co-inhibitory checkpoint is TIGIT (T cell immunoreceptor with Ig and immunoreceptor tyrosine inhibitory motif (ITIM) domains).

[0005] Non-small cell lung cancer (NSCLC) is the major subtype of lung cancer, accounting for approximately 80% - 85% of all cases. In the case of progressive disease, the overall 5-year survival rate is 2% - 4%.

[0006] Despite improvements in the first-line treatment of patients with advanced NSCLC that have resulted in longer survival and a reduction in disease-related symptoms, almost all patients experience disease progression. Cancer immunotherapy, in particular, offers the potential for long-term disease control. In particular, NSCLC patients have been found to benefit from treatment with combinations including the PD-1 axis-binding antagonist (atezolizumab) and the anti-TIGIT antagonist antibody (tiragolumab).

[0007] Accordingly, there is an unmet need in the art for a robust prognostic method to identify patients likely to benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody for more effective management of the disease. SUMMARY OF THE INVENTION

[0008] In one aspect, the invention provides a method of identifying an individual having cancer who is likely to benefit from treatment with 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 therefrom a tumor-associated macrophage (TAM) signature score, wherein an individual having a TAM signature score above a reference TAM signature score is identified as an individual likely to benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0009] In another aspect, the present invention provides a method for selecting a treatment method 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 an individual having a TAM signature score above a reference TAM signature score is identified as an individual who may benefit from a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0010] In some aspects, the individual has a TAM signature score in the sample above the 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.

[0011] In another aspect, the present invention provides a method for treating an individual having cancer, 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 above 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 (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0012] In another aspect, the present invention provides a method of treating an individual having cancer, the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, wherein the individual is determined to have a TAM signature score that exceeds a reference TAM signature score, thereby identifying the individual as an individual who may benefit from treatment with the PD-1 axis-binding antagonist and the 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.

[0013] In some aspects, the sample is obtained from the individual prior to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

[0014] In some aspects, the benefit is an increase in progression-free survival (PFS), objective response rate (ORR), or overall survival (OS).

[0015] In some aspects, the reference TAM signature score is a pre-assigned TAM signature score.

[0016] In some aspects, the reference TAM signature score is the TAM signature score of a reference population. In some aspects, the TAM signature score in the reference population is the median of the TAM signature scores of the reference population. In some aspects, the reference population is a population of individuals having cancer.

[0017] In some embodiments, the TAM signature score is the average of the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from an individual. In some embodiments, the TAM signature score is the average of the normalized expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from an individual.

[0018] In some embodiments, the method further comprises detecting the expression level of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in a sample from an individual.

[0019] In some embodiments, 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 a sample from an individual. In some embodiments, the TAM signature score is the average of the normalized 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 a sample from an individual.

[0020] In some embodiments, the method further comprises detecting the expression level of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in a sample from an 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 a sample from an individual.

[0021] In some embodiments, the expression level of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD has been detected in a sample from an individual.

[0022] In some embodiments, 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 a sample from an individual.

[0023] In some embodiments, the expression level of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD has been detected in a sample from an individual, from which the TAM signature score is determined, and 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 a sample from an individual.

[0024] In another aspect, the present invention provides a method for monitoring the response of an individual having cancer to a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, the method comprising detecting 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 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, 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 a respective reference expression level predicts an individual likely to respond to a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0025] In some aspects, 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 3 weeks after the start of treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0026] In some aspects, 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 6 weeks after the start of treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0027] In some embodiments, 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 level, thereby predicting that the individual is likely to respond to treatment with a PD-1 axis-binding antagonist and an 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.

[0028] In some embodiments, the response to treatment is an increase in PFS or OS.

[0029] In some embodiments, the reference expression level is the baseline expression level from a sample from the individual at a time point prior to the initiation of treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0030] In some embodiments, the present invention provides a method of identifying an individual having cancer who is likely to benefit from treatment with 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 an individual having a Treg signature score that exceeds a reference Treg signature score is identified as an individual who is likely to benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0031] In another aspect, the present invention provides a method for selecting a treatment method 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 an individual having a Treg signature score above a reference Treg signature score is identified as an individual who may benefit from a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0032] In some aspects, the individual has a Treg signature score in the sample above the reference Treg 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.

[0033] In another aspect, the present invention provides a method for treating an individual having cancer, 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 above a reference Treg signature score, whereby the individual is identified 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.

[0034] In another aspect, the present invention is a method of treating an individual having cancer, the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, wherein the individual is determined to have a Treg signature score that exceeds a reference Treg signature score, thereby identifying the individual as an individual who may benefit from treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, and wherein the Treg signature score is based on the expression levels of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 detected in a sample from the individual, and provides a method.

[0035] In some aspects, the sample is obtained from the individual prior to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

[0036] In some aspects, the benefit is an increase in PFS, ORR, or OS.

[0037] In some aspects, the reference Treg signature score is a pre-assigned Treg signature score.

[0038] In some aspects, the reference Treg signature score is the Treg signature score of a reference population. In some aspects, the Treg signature score in the reference population is the median of the Treg signature scores of the reference population. In some aspects, the reference population is a population of individuals having cancer.

[0039] In some aspects, the Treg signature score is the average of the expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual. In some aspects, the Treg signature score is the average of the normalized expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from the individual.

[0040] In some embodiments, the expression level is a nucleic acid expression level or a protein expression level.

[0041] In some embodiments, the expression level is a nucleic acid expression level. In some embodiments, 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.

[0042] In some embodiments, the nucleic acid expression level is an mRNA expression level. In some embodiments, the mRNA expression level is determined by RNA-seq.

[0043] In some embodiments, the expression level is a protein expression level. In some embodiments, the protein expression level is determined by mass spectrometry.

[0044] In some embodiments, the sample is a tissue sample, a tumor sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

[0045] In some embodiments, the sample is a tissue sample. In some embodiments, the tissue sample is a tumor tissue sample. In some embodiments, the tumor tissue sample is a biopsy material.

[0046] In some embodiments, the sample is a serum sample.

[0047] In some embodiments, the sample is an archival sample, a fresh sample, or a frozen sample.

[0048] In some embodiments, the sample has been determined to have a PD-L1 positive tumor cell fraction by immunohistochemistry (IHC) assay.

[0049] In some embodiments, the PD-L1 positive tumor cell fraction is determined by positive staining with an anti-PD-L1 antibody, and the anti-PD-L1 antibody is SP263, 22C3, SP142, or 28-8.

[0050] In some embodiments, when the PD-L1 positive tumor cell fraction is determined by positive staining with the anti-PD-L1 antibody SP263, it is 50% or more. In some embodiments, the PD-L1 positive tumor cell fraction is calculated using the Ventana SP263 IHC assay.

[0051] In some embodiments, when the PD-L1 positive tumor cell fraction is determined by positive staining with the anti-PD-L1 antibody 22C3, it is 50% or more. In some embodiments, the PD-L1 positive tumor cell fraction is calculated using the pharmDx 22C3 IHC assay.

[0052] In some embodiments, the cancer is lung cancer. In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC).

[0053] In some embodiments, 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).

[0054] In some embodiments, the anti-TIGIT antagonist antibody further comprises the following light chain variable region FRs: (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).

[0055] In some embodiments, the anti-TIGIT antagonist antibody further comprises the following heavy chain variable region FRs: (a) FR-H1 comprising the amino acid sequence of X1VQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 11), where X1 is Q or E in the sequence; (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).

[0056] In some embodiments, X1 is Q. In some embodiments, X1 is E.

[0057] In some embodiments, the anti-TIGIT antagonist antibody comprises: (a) a VH domain comprising an amino acid sequence having at least 95% sequence identity with the amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO: 17) or QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO: 18); (b) a VL domain comprising an amino acid sequence having at least 95% sequence identity with the amino acid sequence of DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAWYQQKPGQPPNLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPFTFGPGTKVEIK (SEQ ID NO: 19); or (c) a VH domain as in (a) and a VL domain as in (b).

[0058] In some embodiments, 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.

[0059] In some embodiments, 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.

[0060] In some embodiments, 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.

[0061] In some embodiments, the anti-TIGIT antagonist antibody is a monoclonal antibody.

[0062] In some embodiments, the anti-TIGIT antagonist antibody is a human antibody.

[0063] In some embodiments, the anti-TIGIT antagonist antibody is a full-length antibody.

[0064] In some embodiments, the anti-TIGIT antagonist antibody exhibits effector function.

[0065] In some embodiments, the anti-TIGIT antagonist antibody comprises an Fc domain that can interact with an Fc gamma receptor (FcγR).

[0066] In some embodiments, the anti-TIGIT antagonist antibody is an IgG class antibody. In some embodiments, the anti-IgG antibody is an IgG1 subclass antibody.

[0067] In some embodiments, the anti-TIGIT antagonist antibody is tiragolumab.

[0068] In some embodiments, the anti-TIGIT antagonist antibody is an antibody fragment that binds to TIGIT selected from the group consisting of Fab, Fab’, Fab’-SH, Fv, single-chain variable fragment (scFv), and (Fab’)2 fragments.

[0069] In some embodiments, the anti-TIGIT antagonist antibody is vibostolimab, etigilimab, EOS084448, SGN-TGT, TJ-T6, BGB-A1217, or AB308.

[0070] In some embodiments, 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.

[0071] In some embodiments, the PD-1 axis-binding antagonist is a PD-L1-binding antagonist.

[0072] In some embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to one or more of its ligand binding partners. In some embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1, B7-1, or both PD-1 and B7-1.

[0073] In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antagonist antibody. In some embodiments, the anti-PD-L1 antagonist antibody is atezolizumab, MDX-1105, durvalumab, avelumab, SHR-1316, CS1001, enoblituzumab, TQB2450, ZKAB001, LP-002, CX-072, IMC-001, KL-A167, APL-502, cosibelimab, rodaplimab, FAZ053, TG-1501, BGB-A333, BCD-135, AK-106, LDP, GR1405, HLX20, MSB2311, RC98, PDL-GEX, KD036, KY1003, YBL-007, or HS-636.

[0074] In some embodiments, the anti-PD-L1 antagonist antibody is atezolizumab.

[0075] In some embodiments, 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); (f) an HVR-L3 sequence comprising the amino acid sequence of QQYLYHPAT (SEQ ID NO: 25).

[0076] In some embodiments, 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) comprises the VH domain described in (a) and the VL domain described in (b).

[0077] In some embodiments, 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.

[0078] In some embodiments, 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.

[0079] In some embodiments, the anti-PD-L1 antagonist antibody is a monoclonal antibody.

[0080] In some embodiments, the anti-PD-L1 antagonist antibody is a humanized antibody.

[0081] In some embodiments, the anti-PD-L1 antagonist antibody is a full-length antibody.

[0082] In some embodiments, the anti-PD-L1 antagonist antibody is an antibody fragment that binds to PD-L1 selected from the group consisting of Fab, Fab’, Fab’-SH, Fv, single-chain variable fragment (scFv), and (Fab’)2 fragments.

[0083] In some embodiments, the anti-PD-L1 antagonist antibody is an IgG-class antibody. In some embodiments, the anti-IgG antibody is an IgG1 subclass antibody.

[0084] In some embodiments, the PD-1 axis-binding antagonist is a PD-1-binding antagonist. In some embodiments, the PD-1-binding antagonist inhibits the binding of PD-1 to one or more of its ligand-binding partners. In some embodiments, the PD-1-binding antagonist inhibits the binding of PD-1 to PD-L1, PD-L2, or both PD-L1 and PD-L2.

[0085] In some embodiments, the PD-1-binding antagonist is an anti-PD-1 antagonist antibody. In some embodiments, the anti-PD-1 antagonist antibody is nivolumab, pembrolizumab, MEDI-0680, spartalizumab, semaprilumab, BGB-108, prorgolimab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostarlimab, retyphanlimab, sasanalimab, pemprilumab, CS1003, HLX10, SCT-I10A, zinberelimab, balsilimab, genolimzumab, BI754091, cetrelimab, YBL-006, BAT1306, HX008, budigalimab, AMG404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, AM0001, ENUM 244C8, ENUM 388D4, STI-1110, AK-103 or hAb21.

[0086] In some embodiments, the PD-1-binding antagonist is an Fc fusion protein. In some embodiments, the Fc fusion protein is AMP-224.

[0087] In some embodiments, the individual is human.

[0088] In some embodiments, the anti-TIGIT antagonist antibody is capable of Fc-dependent activation of myeloid cells, and optionally, the myeloid cells are cells selected from the group consisting of intratumoral type 1 conventional dendritic cells (cDC1), macrophages, neutrophils, and circulating monocytes.

[0089] In some embodiments, the anti-TIGIT antagonist antibody can interact with the Fc gamma receptor (FcγR) on myeloid cells and can induce CD8+ T cell mobilization in the blood and / or expansion of proliferating CD8+ T cells within the tumor bed.

[0090] In another aspect, the present invention provides a method of treating an individual having cancer, comprising administering an anti-TIGIT antagonist antibody to the individual, wherein the anti-TIGIT antagonist antibody is capable of Fc-dependent activation of myeloid cells, and optionally, the myeloid cells are cells selected from the group consisting of intratumoral type 1 conventional dendritic cells (cDC1), macrophages, neutrophils, and circulating monocytes.

[0091] In another aspect, the present invention provides the use of an anti-TIGIT antagonist antibody in the manufacture of a medicament for treating cancer, wherein the anti-TIGIT antagonist antibody is capable of Fc-dependent activation of myeloid cells, and optionally, the myeloid cells are cells selected from the group consisting of intratumoral cDC1, macrophages, neutrophils, and circulating monocytes.

[0092] In another aspect, the present invention provides a method of treating an individual having cancer, comprising administering an anti-TIGIT antagonist antibody to the individual, wherein the anti-TIGIT antagonist antibody can interact with the Fc gamma receptor (FcγR) on myeloid cells and can induce mobilization of CD8+ T cells in the blood and / or expansion of proliferating CD8+ T cells within the tumor bed.

[0093] In another aspect, the present invention provides the use of an anti-TIGIT antagonist antibody in the manufacture of a medicament for treating cancer, wherein the anti-TIGIT antagonist antibody can interact with FcγR and can induce mobilization of CD8+ T cells in the blood and / or expansion of proliferating CD8+ T cells within the tumor bed.

[0094] In another aspect, the present invention provides a method for identifying an individual having cancer who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting 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 therefrom a tumor-associated macrophage (TAM) signature score, wherein an individual having a TAM signature score above a reference TAM signature score is identified as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0095] In another aspect, the present invention provides a method for selecting a treatment method 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 therefrom a TAM signature score, wherein an individual having a TAM signature score above a reference TAM signature score is identified as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0096] In some aspects, the individual has a TAM signature score in the sample above the 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 exhibiting effector function.

[0097] In another aspect, the present invention is a method of treating an individual having cancer, 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 exceeds a reference TAM signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, 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; 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.

[0098] In another aspect, the present invention is a method of treating an individual having 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 signature score that exceeds a reference TAM signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting 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.

[0099] In another aspect, the present invention provides a method for monitoring the response of an individual having cancer to a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, the method comprising detecting 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 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 exhibiting effector function, 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 each respective reference expression level predicts an individual likely to respond to a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0100] In another aspect, the present invention provides a method for identifying an individual having cancer who may benefit from a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting 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 above 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 exhibiting effector function.

[0101] In another aspect, the present invention provides a method for selecting a treatment method 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 an individual having a Treg signature score above a reference Treg signature score is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0102] In some aspects, the individual has a Treg signature score in the sample above the reference Treg 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 exhibiting effector function.

[0103] In another aspect, the present invention provides a method for treating an individual having cancer, 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 above a reference Treg signature score, whereby the individual is identified as an individual who may benefit from treatment with 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 a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

[0104] In another aspect, the present invention is a method of 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 exceeds a reference Treg signature score, whereby the individual is identified as an individual who may benefit from treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody that exhibits effector function, and 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, and provides a method.

[0105] In some aspects, the anti-TIGIT antagonist antibody comprises an Fc domain capable of interacting with an Fc gamma receptor (FcγR).

Brief Description of the Drawings

[0106]

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Modes for Carrying Out the Invention

[0107] I. Overview The present invention is based at least in part on the surprising finding that a greater abundance of immunosuppressive cells, particularly tumor-associated macrophages (TAMs) and regulatory T cells (Tregs), is associated with improved objective response rate (ORR), overall survival (OS), and progression-free survival (PFS) for the combination therapy of tirzolgomab + atezolizumab, but not for atezolizumab monotherapy. In particular, analysis of gene expression in tumor samples from patients in the Phase 2 CITYSCAPE study (GO30103) found that median TAM and Treg gene signature scores were each associated with improved outcomes for the combination therapy of tirzolgomab + atezolizumab. Furthermore, analysis of pre-treatment and on-treatment serum samples (Cycle 2, Day 1 (C2D1) and Cycle 3, Day 1 (C3D1)) collected from CITYSCAPE patients showed a statistically significant increase in myeloid-related protein peptides such as MARCO (macrophage receptor with collagenous structure), CSF1R, CD163, CAMP, CD5L, and apolipoproteins (APOC2 / 3 / 4) in the combination treatment arm of tirzolgomab + atezolizumab compared to baseline and 3 weeks post-treatment (C2D1), indicating that myeloid activation is a treatment-specific effect. Surprisingly, it has now also been found that the increase in the levels of these myeloid proteins is associated with longer PFS and OS in patients receiving the combination therapy of tirzolgomab + atezolizumab compared to patients receiving atezolizumab monotherapy for OS in patients with serum myeloid protein increases above the median. Thus, the combination treatment shows a transient increase in serum myeloid proteins, which is differentially associated with improved PFS and OS in the combination treatment arm of tirzolgomab + atezolizumab, indicating that myeloid cells are expected to play an important role in enhancing the antitumor effect of tirzolgomab + atezolizumab.

[0108] Furthermore, in this specification, there is no report on atezolizumab monotherapy including the MYC targeting pathway shown to regulate macrophage polarization, and a novel pathway upregulated in monocytes specific to the tirzgolumab + atezolizumab combination therapy is also discovered.

[0109] Surprisingly, it is now also discovered that the observed myeloid activation requires the tirzgolumab Fc domain interaction with the Fcγ receptor.

[0110] II. General Techniques and Definitions The techniques and procedures described or referenced herein are generally well understood, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual 3d edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Current Protocols in Molecular Biology (F.M. Ausubel, et al. eds., (2003)); Methods in Enzymology series (Academic Press, Inc.): PCR 2: A Practical Approach (M.J. MacPherson, B.D. Hames and G.R. Taylor eds. (1995)), Harlow and Lane, eds. (1988) Antibodies, A Laboratory Manual, and Animal Cell Culture (R.I. Freshney, ed. (1987)); Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney), ed., 1987); Introduction to Cell and Tissue Culture (J.P. Mather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J.B. Griffiths, and D.G. Newell, eds., 1993 - 8) J.Wiley and Sons; Handbook of Experimental Immunology (D.M. Weir and C.C. Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J.M. Miller and M.P.The conventional methodologies such as those widely used methodologies described in Calos, eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Current Protocols in Immunology (J.E. Coligan et al., eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway 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, eds., Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J.D. Capra, eds., Harwood Academic Publishers, 1995); and Cancer: Principles and Practice of Oncology (V.T. DeVita et al., eds., J.B. Lippincott Company, 1993) are commonly used by those skilled in the art.

[0111] It is understood that the aspects and embodiments of the invention described herein include those aspects and embodiments "comprising", "consisting of", and "consisting essentially of". As used herein, the singular forms "a", "an", and "the" include plural referents unless specifically stated otherwise.

[0112] As used herein, the term "about" refers to the normal error range of each value, which can be easily understood by those skilled in the art of this technology. References to values or parameters following "about" in this specification include (and describe) embodiments directed to the value or parameter itself. For example, a description referring to "about X" includes a description of "X".

[0113] As used interchangeably herein, the "amount", "level", or "expression level" of a biomarker is a detectable level in a biological sample. "Expression" generally refers to the process by which information (e.g., gene code information and / or epigenetic information) is present in a cell and converted into a structure that functions therein. Thus, as used herein, "expression" may refer to transcription into a polynucleotide, translation into a polypeptide, or further modification of a polynucleotide and / or polypeptide (e.g., post-translational modification of a polypeptide). A fragment of a transcribed polynucleotide, a translated polypeptide, or a polynucleotide and / or polypeptide modification (e.g., post-translational modification of a polypeptide), whether it is derived from a transcript generated by alternative splicing or a degraded transcript or, for example, from post-translational processing of a polypeptide by proteolysis, should be considered expressed. An "expressed gene" includes those that are transcribed into a polynucleotide as mRNA and then translated into a polypeptide, as well as those that are transcribed into RNA but not translated into a polypeptide (e.g., transfer and ribosomal RNA). Expression levels are known to those skilled in the art and can be measured by methods disclosed herein as well.

[0114] The terms "detect" and "detection" are used herein in the broadest sense to include both qualitative and quantitative measurements of a target molecule. Detection includes simply identifying the presence of a target molecule in a sample and determining whether the target molecule is present in the sample at a detectable level. Detection may be direct or indirect.

[0115] The presence and / or expression level / amount of the various biomarkers described herein in a sample can be analyzed by several methodologies, many of which are known in the art and understood by those of ordinary skill in the art, including immunohistochemistry (“IHC”), Western blot analysis, immunoprecipitation, molecular binding assays, ELISA, ELIFA, fluorescence-activated cell sorting (“FACS”), MassARRAY, proteomics, quantitative blood-based assays (e.g., serum ELISA), biochemical enzyme activity assays, in situ hybridization, fluorescence in situ hybridization (FISH), Southern analysis, Northern analysis, whole genome sequencing, massively parallel DNA sequencing (e.g., next-generation sequencing), NANOSTRING® (e.g., branched DNA, SISBA, TMA, etc.), polymerase chain reaction (PCR) including quantitative real-time PCR (qRT-PCR) and other amplification-based detection methods, RNA-Seq, microarray analysis, gene expression profiling, and / or serial analysis of gene expression (“SAGE”), and / or any one of a variety of assays that can be performed by protein, gene, and / or tissue array analysis, including but not limited to these. Typical protocols for assessing the status of genes and gene products can be found, for example, in Ausubel et al., eds., 1995, Current Protocols In Molecular Biology, Part 2 (Northern blotting), Part 4 (Southern blotting), Part 15 (immunoblotting), and Part 18 (PCR analysis). Multiplexed immunoassays such as those available from Rules Based Medicine or Meso Scale Discovery (“MSD”) can also be used.

[0116] As used herein, the term "complement C1q subcomponent subunit C" or "C1QC" broadly refers to any native C1QC from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length C1QC and isolated regions or domains of C1QC, such as C1QC ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of C1QC that do not affect the function and / or activity of C1QC, are also contemplated by the present invention.

[0117] As used herein, the term "macrophage scavenger receptor type I and type II" or "MSR1" broadly refers to any native MSR1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length MSR1 and isolated regions or domains of MSR1, such as MSR1 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of MSR1 that do not affect the function and / or activity of MSR1, are also contemplated by the present invention.

[0118] As used herein, the term "macrophage mannose receptor 1" or "MRC1" broadly refers to any native MRC1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length MRC1 and isolated regions or domains of MRC1, such as MRC1 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of MRC1 that do not affect the function and / or activity of MRC1, are also contemplated by the present invention.

[0119] As used herein, the term "V-set and immunoglobulin domain-containing protein 4" or "VSIG4" broadly refers to any native VSIG4 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length VSIG4 and isolated regions or domains of VSIG4, such as VSIG4 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of VSIG4 that do not affect the function and / or activity of VSIG4, are also contemplated by the present invention.

[0120] As used herein, the term "secreted phosphoprotein 1" or "SPP1" broadly refers to any native SPP1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length SPP1 and isolated regions or domains of SPP1, such as the SPP1 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of SPP1 that do not affect the function and / or activity of SPP1, are also contemplated by the present invention.

[0121] As used herein, the term "macrophage receptor with a collagenous structure" or "MARCO" broadly refers to any native MARCO from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length MARCO and isolated regions or domains of MARCO, such as the MARCO ECD. This term also encompasses naturally occurring variants of MARCO, such as splice variants or allelic variants. An exemplary amino acid sequence of human MARCO is shown under UniProt accession number Q9UEW3. Minor sequence variations, particularly conservative amino acid substitutions of MARCO that do not affect the function and / or activity of MARCO, are also contemplated by the present invention.

[0122] As used herein, the terms "tartrate-resistant acid phosphatase type 5" or "ACP5" broadly refer to any native ACP5 from any mammalian source, including, but not limited to, primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length ACP5 and isolated regions or domains of ACP5, such as ACP5 ECD. This 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. Minor sequence differences, particularly conservative amino acid substitutions of ACP5 that do not affect the function and / or activity of ACP5, are also contemplated by the present invention.

[0123] As used herein, the terms "mast cell expressed membrane protein 1" or "MCEMP1" broadly refer to any native MCEMP1 from any mammalian source, including, but not limited to, primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length MCEMP1 and isolated regions or domains of MCEMP1, such as MCEMP1 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of MCEMP1 that do not affect the function and / or activity of MCEMP1, are also contemplated by the present invention.

[0124] As used herein, the term "sterol 27-hydroxylase" or "CYP27A1" broadly refers to any native CYP27A1 from any mammalian source, including but not limited to primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length CYP27A1 and isolated regions or domains of CYP27A1, such as the CYP27A1 ECD. This term also includes 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. Minor sequence variations, particularly conservative amino acid substitutions of CYP27A1 that do not affect the function and / or activity of CYP27A1, are also contemplated by the present invention.

[0125] As used herein, the term "oxidized low density lipoprotein receptor 1" or "OLR1" broadly refers to any native OLR1 from any mammalian source, including but not limited to primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length OLR1 and isolated regions or domains of OLR1, such as the OLR1 ECD. This term also includes 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. Minor sequence variations, particularly conservative amino acid substitutions of OLR1 that do not affect the function and / or activity of OLR1, are also contemplated by the present invention.

[0126] As used herein, the terms "progranulin" or "GRN" broadly refer to any native GRN from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length GRN and isolated regions or domains of GRN, such as GRN ECD. This term also encompasses naturally occurring variants of GRN, such as splice variants or allelic variants. An exemplary amino acid sequence of human GRN is shown under UniProt accession number P28799. Minor sequence variations, particularly conservative amino acid substitutions of GRN that do not affect the function and / or activity of GRN, are also contemplated by the present invention.

[0127] As used herein, the terms "glioma pathogenicity-related protein 2" or "GLIPR2" broadly refer to any native GLIPR2 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length GLIPR2 and isolated regions or domains of GLIPR2, such as GLIPR2 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of GLIPR2 that do not affect the function and / or activity of GLIPR2, are also contemplated by the present invention.

[0128] As used herein, the term "arrestin domain-containing protein 4" or "ARRDC4" broadly refers to any native ARRDC4 from any mammalian source, including, without limitation, primates (e.g., humans) and rodents (e.g., mice and rats). This term encompasses full-length ARRDC4 and isolated regions or domains of ARRDC4, such as ARRDC4 ECD. This 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. Minor sequence differences, particularly conservative amino acid substitutions of ARRDC4 that do not affect the function and / or activity of ARRDC4, are also contemplated by the present invention.

[0129] As used herein, the term "apolipoprotein E" or "APOE" broadly refers to any native APOE from any mammalian source, including, without limitation, primates (e.g., humans) and rodents (e.g., mice and rats). This term encompasses full-length APOE and isolated regions or domains of APOE, such as APOE ECD. This 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. Minor sequence differences, particularly conservative amino acid substitutions of APOE that do not affect the function and / or activity of APOE, are also contemplated by the present invention.

[0130] As used herein, the term "folate receptor beta" or "FOLR2" broadly refers to any native FOLR2 from any mammalian source, including, without limitation, primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length FOLR2 and isolated regions or domains of FOLR2, such as FOLR2 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of FOLR2 that do not affect the function and / or activity of FOLR2, are also contemplated by the present invention.

[0131] As used herein, the term "cathepsin D" or "CTSD" broadly refers to any native CTSD from any mammalian source, including, without limitation, primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length CTSD and isolated regions or domains of CTSD, such as CTSD ECD. This term also encompasses naturally occurring variants of CTSD, such as splice variants or allelic variants. An exemplary amino acid sequence of human CTSD is shown under UniProt accession number P07339. Minor sequence variations, particularly conservative amino acid substitutions of CTSD that do not affect the function and / or activity of CTSD, are also contemplated by the present invention.

[0132] As used herein, the term "cathelicidin antimicrobial peptide" or "cAMP" broadly refers to any native cAMP from any mammalian source, including, but not limited to, primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length CAMP and isolated regions or domains of CAMP, such as CAMP ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of CAMP that do not affect the function and / or activity of CAMP, are also contemplated by the present invention.

[0133] As used herein, the term "CD5 antigen-like" or "CD5L" broadly refers to any native CD5L from any mammalian source, including, but not limited to, primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length CD5L and isolated regions or domains of CD5L, such as CD5L ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of CD5L that do not affect the function and / or activity of CD5L, are also contemplated by the present invention.

[0134] As used herein, the term "scavenger receptor cysteine-rich type 1 protein M130" or "CD163" broadly refers to any native CD163 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length CD163 and isolated regions or domains of CD163, such as CD163 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of CD163 that do not affect the function and / or activity of CD163, are also contemplated by the present invention.

[0135] As used herein, the term "neutrophil gelatinase-associated lipocalin" or "NGAL" broadly refers to any native NGAL from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length NGAL and isolated regions or domains of NGAL, such as NGAL ECD. This term also encompasses naturally occurring variants of NGAL, such as splice variants or allelic variants. An exemplary amino acid sequence of human NGAL is shown under UniProt accession number P80188. Minor sequence variations, particularly conservative amino acid substitutions of NGAL that do not affect the function and / or activity of NGAL, are also contemplated by the present invention.

[0136] As used herein, the term "macrophage colony-stimulating factor 1 receptor" or "CSF1R" broadly refers to any native CSF1R from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length CSF1R and isolated regions or domains of CSF1R, such as CSF1R ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of CSF1R that do not affect the function and / or activity of CSF1R, are also contemplated by the present invention.

[0137] As used herein, the term "CD44 antigen" or "CD44" broadly refers to any native CD44 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length CD44 and isolated regions or domains of CD44, such as CD44 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of CD44 that do not affect the function and / or activity of CD44, are also contemplated by the present invention.

[0138] As used herein, the term "apolipoprotein C-II" or "APOC2" broadly refers to any native APOC2 from any mammalian source, including but not limited to primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length APOC2 and isolated regions or domains of APOC2, such as the APOC2 ECD. This 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. Minor sequence differences, particularly conservative amino acid substitutions of APOC2 that do not affect the function and / or activity of APOC2, are also contemplated by the present invention.

[0139] As used herein, the term "apolipoprotein C-III" or "APOC3" broadly refers to any native APOC3 from any mammalian source, including but not limited to primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length APOC3 and isolated regions or domains of APOC3, such as the APOC3 ECD. This 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. Minor sequence differences, particularly conservative amino acid substitutions of APOC3 that do not affect the function and / or activity of APOC3, are also contemplated by the present invention.

[0140] As used herein, the terms "apolipoprotein C-IV" or "APOC4" broadly refer to any native APOC4 from any mammalian source, including, but not limited to, primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length APOC4 and isolated regions or domains of APOC4, such as APOC4 ECD. This 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. Minor sequence differences, particularly conservative amino acid substitutions of APOC4 that do not affect the function and / or activity of APOC4, are also contemplated by the present invention.

[0141] As used herein, the terms "apolipoprotein A-II" or "APOA2" broadly refer to any native APOA2 from any mammalian source, including, but not limited to, primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length APOA2 and isolated regions or domains of APOA2, such as APOA2 ECD. This 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. Minor sequence differences, particularly conservative amino acid substitutions of APOA2 that do not affect the function and / or activity of APOA2, are also contemplated by the present invention.

[0142] As used herein, the terms "lactotransferrin" or "TRFL" broadly refer to any native TRFL from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length TRFL and isolated regions or domains of TRFL, such as TRFL ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of TRFL that do not affect the function and / or activity of TRFL, are also contemplated by the present invention.

[0143] As used herein, the terms "vascular cell adhesion protein 1" or "VCAM1" broadly refer to any native VCAM1 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length VCAM1 and isolated regions or domains of VCAM1, such as VCAM1 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of VCAM1 that do not affect the function and / or activity of VCAM1, are also contemplated by the present invention.

[0144] As used herein, the terms "beta-2 microglobulin" or "B2MG" broadly refer to any native B2MG from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length B2MG and isolated regions or domains of B2MG, such as B2MG ECD. This term also encompasses naturally occurring variants of B2MG, such as splice variants or allelic variants. An exemplary amino acid sequence of human B2MG is shown under UniProt accession number P61769. Minor sequence variations, particularly conservative amino acid substitutions of B2MG that do not affect the function and / or activity of B2MG, are also contemplated by the present invention.

[0145] As used herein, the terms "forkhead box protein P3" or "FOXP3" broadly refer to any native FOXP3 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length FOXP3 and isolated regions or domains of FOXP3, such as FOXP3 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of FOXP3 that do not affect the function and / or activity of FOXP3, are also contemplated by the present invention.

[0146] As used herein, the term "cytotoxic T-lymphocyte protein 4" or "CTLA4" broadly refers to any native CTLA4 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length CTLA4 and isolated regions or domains of CTLA4, such as CTLA4 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of CTLA4 that do not affect the function and / or activity of CTLA4, are also contemplated by the present invention.

[0147] As used herein, the term "interleukin 10" or "IL10" broadly refers to any native IL10 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length IL10 and isolated regions or domains of IL10, such as IL10 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of IL10 that do not affect the function and / or activity of IL10, are also contemplated by the present invention.

[0148] As used herein, the term "tumor necrosis factor receptor superfamily member 18" or "TNFRSF18" broadly refers to any native TNFRSF18 from any mammalian source, including, without limitation, primates (e.g., humans) and rodents (e.g., mice and rats). This term encompasses full-length TNFRSF18 and isolated regions or domains of TNFRSF18, such as TNFRSF18 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of TNFRSF18 that do not affect the function and / or activity of TNFRSF18, are also contemplated by the present invention.

[0149] As used herein, the term "C-C chemokine receptor type 8" or "CCR8" broadly refers to any native CCR8 from any mammalian source, including, without limitation, primates (e.g., humans) and rodents (e.g., mice and rats). This term encompasses full-length CCR8 and isolated regions or domains of CCR8, such as CCR8 ECD. This 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. Minor sequence variations, particularly conservative amino acid substitutions of CCR8 that do not affect the function and / or activity of CCR8, are also contemplated by the present invention.

[0150] As used herein, the term "zinc finger protein Eos (Eos)" or "IKZF4" broadly refers to any native IKZF4 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length IKZF4 and isolated regions or domains of IKZF4, such as IKZF4 ECD. This term also encompasses naturally occurring variants of IKZF4, such as splice variants or allelic variants. An exemplary human IKZF4 amino acid sequence is shown under UniProt accession number Q9H2S9. Minor sequence variations, particularly conservative amino acid substitutions of IKZF4 that do not affect the function and / or activity of IKZF4, are also contemplated by the present invention.

[0151] As used herein, the term "zinc finger protein Helios (Helios)" or "IKZF2" broadly refers to any native IKZF2 from any mammalian source, including primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. This term encompasses full-length IKZF2 and isolated regions or domains of IKZF2, such as IKZF2 ECD. This term also encompasses naturally occurring variants of IKZF2, such as splice variants or allelic variants. An exemplary human IKZF2 amino acid sequence is shown under UniProt accession number Q9UKS7. Minor sequence variations, particularly conservative amino acid substitutions of IKZF2 that do not affect the function and / or activity of IKZF2, are also contemplated by the present invention.

[0152] As used herein, "TIGIT" or "T-cell immunoreceptor with Ig and ITIM domains" refers to any native TIGIT from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. TIGIT is also known in the art as DKFZp667A205, FLJ39873, V-set and immunoglobulin domain-containing protein 9, V-set and transmembrane domain-containing protein 3, VSIG9, VSTM3, and WUCAM. This term encompasses "full-length" unprocessed TIGIT (e.g., full-length human TIGIT having the amino acid sequence of SEQ ID NO: 30), as well as any form of TIGIT resulting from intracellular processing (e.g., processed human TIGIT without a signal sequence having the amino acid sequence of SEQ ID NO: 31). This term also encompasses naturally occurring variants of TIGIT, such as splice variants or allelic variants. An exemplary amino acid sequence of human TIGIT can be found, for example, under UniProt accession number Q495A1.

[0153] The term "PD-L1" or "programmed cell death ligand 1" as used herein refers to any native PD-L1 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. PD-L1 is also known in the art as CD274 molecule, CD274 antigen, B7 homolog 1, PDCD1 ligand 1, PDCD1LG1, PDCD1L1, B7H1, PDL1, programmed death ligand 1, B7-H1 and B7-H. This term also encompasses naturally occurring variants of PD-L1, 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).

[0154] The term "antagonist" is used in the broadest sense and includes any molecule that partially or completely blocks, inhibits, or neutralizes the biological activity of the native 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 native polypeptides, peptides, antisense oligonucleotides, small organic molecules, and the like. Methods for identifying polypeptide antagonists can include contacting the polypeptide with a candidate antagonist molecule and measuring a detectable change in one or more biological activities normally associated with the polypeptide.

[0155] The term "PD-1 axis-binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis-binding partner with any one or more of its binding partners so as to remove T cell dysfunction resulting from signal transduction on the PD-1 signaling axis, and as a result, restores or enhances 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.

[0156] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, impairs, or interferes with the signaling resulting from the interaction of PD-1 with one or more binding partners such as PD-L1, PD-L2, etc. In some embodiments, the PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to one or more of its binding partners. In a specific embodiment, the 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, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, impair, or interfere with the signaling resulting from the interaction of PD-1 with PD-L1 and / or PD-L2. In one embodiment, the PD-1 binding antagonist reduces the negative co-stimulatory signal mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-1, and makes the dysfunctionality of dysfunctional T cells lower (e.g., enhances the effector response to antigen recognition). In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In a specific embodiment, the PD-1 binding antagonist is MDX-1106 (nivolumab) described herein. In another specific embodiment, the PD-1 binding antagonist is pembrolizumab (previously lambrolizumab (MK-3475)) described herein. In another specific embodiment, the PD-1 binding antagonist is AMP-224 described herein.

[0157] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction resulting from the interaction of PD-L1 with one or more of its binding partners, such as PD-1, B7-1. In some embodiments, the PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In a specific embodiment, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, the PD-L1 binding antagonist includes anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abolish, or interfere with signal transduction resulting from the interaction of PD-L1 with one or more of its binding partners, such as PD-1, B7-1. In one embodiment, the PD-L1 binding antagonist reduces the negative co-stimulatory signal mediated by or through a cell surface protein expressed in response to signal transduction mediated by T lymphocytes via PD-L1, and reduces the dysfunction of dysfunctional T cells (e.g., enhances the effector response to antigen recognition). In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In a specific embodiment, the anti-PD-L1 antibody is atezolizumab (e.g., MPDL3280A) as described herein. In another specific embodiment, the anti-PD-L1 antibody is MDX-1105 as described herein. In yet another specific embodiment, the anti-PD-L1 antibody is MEDI4736 as described herein.

[0158] As used herein, the term "atezolizumab" refers to an anti-PD-L1 antagonist antibody having the International Nonproprietary Name (INN) list 112 (WHO Drug Information, Vol. 28, No. 4, 2014, p. 488) or CAS registration number 1380723-44-3.

[0159] The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In some embodiments, the PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to one or more of its binding partners. In a specific embodiment, the PD-L2 binding antagonist inhibits the binding of PD-L2 to PD-1. In some embodiments, the PD-L2 antagonist includes an anti-PD-L2 antibody, an antigen-binding fragment thereof, an immunoadhesin, a fusion protein, an oligopeptide, and other molecules that reduce, block, inhibit, suppress, or interfere with signal transduction resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In one embodiment, the PD-L2 binding antagonist reduces the negative co-stimulatory signal mediated by or through a cell surface protein expressed in response to T lymphocyte-mediated signal transduction via PD-L2, and alleviates the dysfunction of dysfunctional T cells (e.g., enhances the effector response to antigen recognition). In some embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0160] The term "anti-TIGIT antagonist antibody" refers to an antibody, or antigen-binding fragment or variant thereof, 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 to restore the functional response to antigenic stimulation from a dysfunctional state (e.g., proliferation, cytokine production, target cell killing) by T cells. For example, an anti-TIGIT antagonist antibody can block signaling via PVR without affecting the PVR-CD226 interaction. In some examples, it will be understood by those skilled in the art that an anti-TIGIT antagonist antibody can antagonize one TIGIT activity without affecting another TIGIT activity. For example, an anti-TIGIT antagonist antibody for use or use in the particular methods described herein is an anti-TIGIT antagonist antibody that antagonizes TIGIT activity in response to one of a PVR interaction, a PVRL3 interaction, or a PVRL2 interaction, for example, without affecting or minimally affecting any of the other TIGIT interactions. In one embodiment, the degree of binding of the anti-TIGIT antagonist antibody to an unrelated non-TIGIT protein is less than about 10% of the binding of the antibody to TIGIT as measured, for example, by radioimmunoassay (RIA). In certain embodiments, the TIGIT to which the anti-TIGIT antagonist antibody binds has a dissociation constant (K -8 ) of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., 10 -8 M or less, e.g., 10 -13 M to 10 -9 M, e.g., 10 -13 M to 10- D M). In certain embodiments, the anti-TIGIT antagonist antibody binds to an epitope of TIGIT that is conserved among TIGITs from different species or an epitope on TIGIT that permits cross-species reactivity. In one embodiment, the anti-TIGIT antagonist antibody is tirzolgomab.

[0161] As used herein, "tilagolumab" is a fully human IgG1 / kappa MAb derived from Open Monoclonal Technology (OMT) rats that binds to TIGIT and comprises the heavy chain sequence of SEQ ID NO: 33 and the light chain sequence of SEQ ID NO: 34. Tilagolumab contains two N-linked glycosylation sites (N306) in the Fc domain. Tilagolumab is also described in WHO Drug Information (International Nonproprietary Name for Pharmaceutical Substances), Proposed INN: List 117, Vol. 31, No. 2 (issued June 9, 2017) (see page 343).

[0162] As used herein, "administering" means a method of giving a dosage 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, e.g., a pharmaceutical composition comprising an anti-TIGIT antibody and / or a PD-1 axis-binding antagonist (e.g., an anti-PD-L1 antibody)) to a subject. The compounds and / or compositions used in the methods described herein can be administered, for example, intravenously (e.g., by intravenous infusion), subcutaneously, intramuscularly, intradermally, transdermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, rectally, topically, intratumorally, intraperitoneally, subconjunctivally, intravesicularly, transmucosally, intrapericardially, intraumbilically, intraocularly, orally, topically, locally, by inhalation, injection, infusion, continuous infusion, local perfusion directly flowing to target cells, catheter, lavage, cream, or lipid composition. The method of administration can vary depending on various factors (e.g., the compound or composition being administered and the severity of the condition, disease, or disorder being treated).

[0163] As used herein, "systemic treatment" refers to a treatment that moves through the bloodstream and can contact multiple organ systems upon a single administration. The term "systemic treatment" is well understood by those skilled in the art and is equivalent to systemic therapy.

[0164] As used herein, the "fixed" or "flat" dose of a therapeutic agent (e.g., an anti-TIGIT antagonist antibody or a PD-1 axis-binding antagonist (e.g., an anti-PD-L1 antibody)) refers to the dose administered to a patient regardless of the patient's weight or body surface area (BSA). Thus, a fixed dose or flat dose is provided as the absolute amount of the therapeutic agent (e.g., mg), rather than as a mg / kg dose or mg / m 2 dose.

[0165] As used herein, the terms "treatment" or "treating" refer to a clinical intervention designed to alter the natural course of an individual or cell being treated during the course of a clinical pathology. Desirable effects of treatment include delaying or reducing the rate of disease progression, restoring or alleviating the disease state, and remission or improvement of the prognosis. For example, an individual has a reduction in the growth (or destruction) of cancerous cells, a decrease in symptoms resulting from the disease, an improvement in the quality of life of those suffering from the disease, a decrease in the dosage of other pharmaceuticals required to treat the disease, a delay in the progression of the disease, and / or an extension of the individual's survival period, including but not limited to, when one or more symptoms associated with cancer are reduced or eliminated, "treatment" is considered successful.

[0166] As used herein, "in combination with" refers to the application of one treatment modality added to another treatment modality. Thus, "in combination with" refers to the application of another treatment modality before, during, or after the application of one treatment modality to an individual.

[0167] A "disorder" or "disease" is any condition that benefits from treatment, including but not limited to disorders associated with some degree of abnormal cell growth, such as cancer, such as lung cancer, such as non-small cell lung cancer (NSCLC).

[0168] As used in connection with immune dysfunction, the term "dysfunction" refers to a state in which the immune responsiveness to antigenic stimulation is reduced.

[0169] As used herein, the term "dysfunctionality" also includes unresponsiveness or non-responsiveness to antigen recognition, particularly impairment of the ability to convert antigen recognition into downstream T cell effector functions such as proliferation, cytokine production (e.g., gamma interferon), and / or target cell killing.

[0170] The terms "cancer" and "cancerous" refer to or describe physiological conditions in mammals typically characterized by uncontrolled cell growth. Examples of cancer include, but are not limited to, carcinomas, lymphomas, blastomas, sarcomas, and leukemias, or lymphoid malignancies. More specific examples of such cancers include, but are not limited to, lung cancers such as non-small cell lung cancer (NSCLC), which includes squamous NSCLC or non-squamous NSCLC, locally advanced unresectable NSCLC (e.g., stage IIIB NSCLC), or recurrent or metastatic NSCLC (e.g., stage IV NSCLC), adenocarcinoma of the lung or squamous cell carcinoma of the lung (e.g., squamous cell carcinoma of the lung), and small cell lung cancer (SCLC) (including extensive-stage SCLC (ES-SCLC)).Further examples of cancer include cancers of the stomach, including gastric cancer, gastrointestinal stromal tumor, 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-resistant non-muscle-invasive bladder cancer (NMIBC)); urinary tract cancer; liver cancer; breast cancer (e.g., HER2+ breast cancer and triple-negative breast cancer (TNBC) that are estrogen receptor (ER-), progesterone receptor (PR-), and HER2 (HER2-) negative); endometrial carcinoma or uterine carcinoma; salivary gland carcinoma; kidney cancer or renal cancer (e.g., renal cell carcinoma (RCC)); prostate cancer, vulvar cancer, thyroid cancer, liver carcinoma, anal carcinoma, penile carcinoma, melanoma (including superficial spreading melanoma, lentigo maligna melanoma, acral lentiginous melanoma, and nodular melanoma), multiple myeloma, and B-cell lymphoma (including low-grade / follicular non-Hodgkin lymphoma (NHL), small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-cleaved cell NHL, large lesion NHL, mantle cell lymphoma, AIDS-related lymphoma, and Waldenström macroglobulinemia), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), hairy cell leukemia, chronic myelogenous leukemia (CML), post-transplant lymphoproliferative disorder (PTLD), and myelodysplastic syndrome (MDS), as well as abnormal blood vessel growth associated with phakomatosis, edema (such as that associated with brain tumors), Meigs syndrome, brain cancer, head and neck cancer, and related metastases.

[0171] The term "tumor" refers to the growth and proliferation of all neoplastic cells, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor," as used herein, are not mutually exclusive.

[0172] "Tumor immunity" refers to the process by which tumors avoid immune recognition and clearance. Thus, as a therapeutic concept, tumor immunity is "treated" when such avoidance is attenuated and the tumor is recognized and attacked by the immune system. Examples of tumor recognition include tumor binding, tumor shrinkage, and tumor clearance.

[0173] As used herein, "metastasis" means the spread of cancer from its primary site to other locations in the body. Cancer cells may break away from the primary tumor, penetrate into lymphatic and blood vessels, circulate through the bloodstream, and grow (metastasize) at distant foci within normal tissues in other parts of the body. Metastasis can be local or distant. Metastasis is a sequential process that requires cancer cells to break away from the primary tumor, move through the bloodstream, and stop at a distant site. At the new site, the cells can establish a blood supply, grow, and form life-threatening masses. Both stimulatory and inhibitory molecular pathways within cancer cells control this behavior, and interactions between cancer cells and host cells at distant sites are also important.

[0174] The term "anti-cancer therapy" refers to a treatment method useful for treating cancer (e.g., lung cancer, e.g., NSCLC). Examples of anti-cancer therapeutic agents include, but are not limited to, for example, immunomodulators (e.g., immunomodulators (e.g., one or more immune co-inhibitory receptors (e.g., one or more immune co-inhibitory receptors selected from TIGIT, PD-L1, PD-1, CTLA-4, LAG3, TIM3, BTLA, and / or VISTA) reducing or inhibiting agents), CTLA-4 antagonists, etc., for example, anti-CTLA-4 antagonist antibodies (e.g., ipilimumab (YERVOY®)), anti-TIGIT antagonist antibodies, or PD-1 axis-binding antagonists (e.g., anti-PD-L1 antibodies), or 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) increasing or activating agents, OX-40 agonists, etc., for example, OX-40 agonist antibodies), chemotherapeutic agents, growth inhibitors, cytotoxic agents, agents used in radiotherapy, anti-angiogenic agents, apoptosis agents, anti-tubulin agents, and other agents for treating cancer. Combinations thereof are also included in the present invention.

[0175] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or blocks the function of cells and / or causes cell death or destruction. Cytotoxic agents include radioisotopes (e.g., At 211 、I 131 、I 125 、Y 90 、Re 186 、Re 188 、Sm 153 、Bi 212 、P 32 、Pb 212, and radioisotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitors; enzymes and fragments thereof, such as nucleolytic enzymes; antibiotics; toxins such as low molecular weight toxins or enzymatically active toxins derived from bacteria, fungi, plants or animals (including fragments and / or variants thereof); and various antitumor agents or anticancer agents disclosed below, but not limited thereto.

[0176] "Chemotherapeutic agents" include chemical compounds useful for the treatment of cancer. Examples of chemotherapeutic agents include erlotinib (TARCEVA®, Genentech / OSI Pharm), bortezomib (VELCADE®, Millennium Pharm.), disulfiram, epigallocatechin gallate, salinosporamide A, carfilzomib, 17-AAG (geldanamycin), radicicol, lactate dehydrogenase A (LDH-A), fulvestrant (FASLODEX®, AstraZeneca), sunitinib (SUTENT®, Pfizer / Sugen), letrozole (FEMARA®, Novartis), imatinib mesylate (GLEEVEC®, Novartis), finasanate (VATALANIB®, Novartis), oxaliplatin (ELOXATIN®, Sanofi), 5-FU (5-fluorouracil), leucovorin, rapamycin (sirolimus, RAPAMUNE®, Wyeth), lapatinib (TYKERB®, GSK572016, Glaxo Smith Kline), lonafarnib (SCH 66336), sorafenib (NEXAVAR®, Bayer Labs), gefitinib (IRESSA®, AstraZeneca), AG1478, thiotepa, and alkylating agents such as CYTOXAN® cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carbochromen, meturedopa, and uredopa; ethyleneimines and methylmelamines such as altretamine, triethylenemelamine, triethylenephosphamide, triethylenethiophosphamide, and trimethylmelamine; acetogenins (especially bullatacin and bullatacinone); camptothecins (topotecan and irinotecan); bryostatin; calicheamicin; CC-1065 (including its synthetic analogs adozelesin, carzelesin, and bizelesin); cryptophycins (especially cryptophycin 1 and cryptophycin 8); corticosteroids (including prednisone and prednisolone); cyproterone acetate;5α-reductases including finasteride and dutasteride; vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat, dacostatins; aldoxorubicin, talarozole, duocarmycin (synthetic analogs including KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin, chlorambucil, chromomycin, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobenbiquine, phenesterine, prednimustine, trofosfamide, uracil mustard and other nitrogen mustards; carmustine, chloroozotocin, fotemustine, lomustine, nimustine, and ranimustine and other nitrosoureas; antibiotics such as enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin γ1I and calicheamicin ω1I (Angew Chem.Intl.Ed.Engl. 1994 33:183-186); dynemicin including dynemicin A; bisphosphonates such as clodronate; esperamicin; similarly, neocarzinostatin chromophore and related chromophores enediyne antibiotic chromophores), aclacinomycins, actinomycins, authramycin, azaserine, bleomycin, cactinomycin, carabicin, caminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, adriamycin (registered trademark) (doxorubicin), morpholinodoxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolinodoxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, keramycin, rhodomycin, streptozocin, streptonigrin, tubercidin, ubenimex, dinostatin, zorubicin; metabolic antagonists such as methotrexate and 5-fluorouracil (5-FU); folate analogs such as denopterin, methotrexate, pteropterin, trimetrexate;Purine analogs such as fludarabine, 6-mercaptopurine, thiampurine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, didoxuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, drostanolone propionate, epithiostanol, mepithiostane, testolactone; antiadrenal agents such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as folic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; dexamethasone; diaziquone; elfomithine; elliptinium acetate; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK (registered trademark) polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2’,2”-trichloroethylamine; trichothecenes (especially T-2 toxin, verrucarin A, roridin A and anguidine); urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids, for example, taxol (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE (registered trademark) (without cremophor), albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE (registered trademark) (paclitaxel; Sanofi-Aventis): chlorambucil, GEMZAR (registered trademark) (gemcitabine), 6-thioguanine, mercaptopurine; methotrexate;Platinum analogs such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE (registered trademark) (vinorelbine); nobandron; teniposide; edatrexate; daunomycin; aminopterin; capecitabine (XELODA (registered trademark)); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid, and pharmaceutically acceptable salts, acids and derivatives of any of the above are mentioned.;

[0177] In addition, as chemotherapeutic agents, (i) antihormonal agents that act to regulate or inhibit the hormonal action on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), for example, tamoxifen (including NOLVADEX (registered trademark), tamoxifen citrate), raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and FARESTON (registered trademark) (toremifene citrate); (ii) aromatase inhibitors that inhibit aromatase, an enzyme that regulates estrogen production in the adrenal glands, for example, 4(5)-imidazole, aminoglutethimide, MEGASE (registered trademark) (megestrol acetate), AROMASIN (registered trademark) (exemestane; Pfizer), formestane, fadrozole, RIVISOR (registered trademark) (vorozole), FEMARA (registered trademark) (letrozole; Novartis), and ARIMIDEX (registered trademark) (anastrozole; AstraZeneca); (iii) antiandrogenic agents such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; buserelin, tripterelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all-trans retinoic acid, fenretinide, and troxacitabine (1,3-dioxolane nucleoside cytosine analog); (iv) protein kinase inhibitors (for example, anaplastic lymphoma kinase (Alk) inhibitors such as AF-802 (also known as CH-5424802 or alectinib)); (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly agents that inhibit the expression of genes in signal transduction pathways involved in abnormal cell proliferation, for example, PKC-alpha, Ralf, and H-Ras; (vii) ribozymes such as VEGF expression inhibitors (for example, ANGIOZYME (registered trademark)), HER2 expression inhibitors, etc.; (viii) vaccines such as gene therapy vaccines, for example, ALLOVECTIN (registered trademark), LEUVECTIN (registered trademark), VAXID (registered trademark); PROLEUKIN (registered trademark), rIL-2; topoisomerase 1 inhibitors such as LURTOTECAN (registered trademark);ABARELIX (registered trademark) rmRH; and pharmaceutically acceptable salts, acids and derivatives of any of the foregoing.

[0178] Chemotherapeutic agents include antibodies such as alemtuzumab (Campath), bevacizumab (AVASTIN®, Genentech), cetuximab (ERBITUX®, Imclone), panitumumab (VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech / Biogen Idec), pertuzumab (OMNITARG®, 2C4, Genentech), trastuzumab (HERCEPTIN®, Genentech), tositumomab (Bexxar, Corixia), and antibody-drug conjugates such as gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Additional humanized monoclonal antibodies having therapeutic potential as agents in combination with the compounds of the present invention include apolizumab, aselizumab, atorizumab, bapineuzumab, bevacizumab mertansine, canertinib mertansine, cedelizumab, certolizumab pegol, cidfostuxizumab, cidotuzumab, daclizumab, eclizumab, efalizumab, epratuzumab, elotuzumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, labesimab, lintuzumab, matuzumab, mapolizumab, motavizumab, motovizumab, natalizumab, nimotuzumab, norovizumab, numavizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pecificumab, pecilizumab, pecilizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab, reslizumab, rovelizumab, rupizumab, sibrotuzumab, siprilizumab, sonotuzumab, takatuzumab tetraxetan, tadoxizumab, talizumab, tefibazumab, tocilizumab, tralizumab, tucotuzumab celmoleukin, tucusizumab, umavizumab, ultoxizumab, ustekinumab, visilizumab, and anti-interleukin-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories), a full-length IgG1λ antibody of only human sequence that has been genetically engineered to recognize interleukin-12 p40 protein.

[0179] A chemotherapeutic agent refers to a compound that binds to or otherwise directly interacts with it and blocks or reduces its signaling activity, and alternatively, an "EGFR inhibitor", also referred to as an "EGFR antagonist". Examples of such agents include antibodies and small molecules that bind to EGFR. Examples of antibodies that bind to EGFR include MAb579 (ATCC CRL HB8506), MAb455 (ATCC CRL HB8507), MAb225 (ATCC CRL8508), MAb528 (ATCC CRL8509) (see U.S. Patent No. 4,943,533, Mendelsohn et al.) and variants thereof, such as chimerized 225 (C225 or cetuximab, ERBUTIX®) and recombinant human 225 (H225) (see International Publication No. 96 / 40210, Imclone Systems Inc.), IMC-11F8, a fully human EGFR-targeting antibody (Imclone), an antibody that binds to type II mutant EGFR (U.S. Patent No. 5,212,290), humanized and chimeric antibodies that bind to EGFR as described in U.S. Patent No. 5,891,996, and human antibodies that bind to EGFR such as ABX-EGF or panitumumab (see International Publication No. 98 / 50433, Abgenix / Amgen), EMD55900 (see Stragliotto et al. Eur. J. Cancer 32A:636-640 (1996)), EMD7200 (matuzumab), a humanized EGFR antibody against EGFR that competes for EGFR binding of both EGF and TGF-alpha (EMD / Merck), a human EGFR antibody, HuMax-EGFR (GenMab), E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, and E7.6.3, fully human antibodies described in U.S. Patent No. 6,235,883, MDX-447 (Medarex Inc), and mAb806 or humanized mAb806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)).An anti-EGFR antibody can be conjugated to a cytotoxic agent, thereby generating an immunoconjugate (see, for example, European Patent Application Publication No. 659,439 A2, Merck Patent GmbH). Examples of EGFR antagonists include the compounds described 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, 5,866,572, 6,399,602, 6,344,459, 6,602,863, 6,391,874, 6,344,455, 5,760,041, 6,002,008, and 5,747,498, and the following PCT publications: International Publication Nos. 98 / 14451, 98 / 50038, 99 / 09016, and 99 / 24037, and small molecules such as those described therein.Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, TARCEVA (registered trademark), Genentech / OSI Pharmaceuticals); PD183805 (CI1033, 2-propenamide, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[3-(4-morpholinyl)propoxy]-6-quinazolinyl]-, dihydrochloride, Pfizer Inc.); ZD1839, gefitinib (IRESSA (registered trademark)) 4-(3'-chloro-4'-fluoroanilino)-7-methoxy-6-(3-morpholinopropoxy)quinazoline, AstraZeneca); ZM105180 ((6-amino-4-(3-methylphenyl-amino)-quinazoline, Zeneca); BIBX-1382 (N8-(3-chloro-4-fluoro-phenyl)-N2-(1-methyl-piperidin-4-yl)-pyrimido[5,4-d]pyrimidine-2,8-diamine, Boehringer Ingelheim); PKI-166 ((R)-4-[4-[(1-phenylethyl)amino]-1H-pyrrolo[2,3-d]pyrimidin-6-yl]-phenol); (R)-6-(4-hydroxyphenyl)-4-[(1-phenylethyl)amino]-7H-pyrrolo[2,3-d]pyrimidine); CL-387785 (N-[4-[(3-bromophenyl)amino]-6-quinazolinyl]-2-butynamide); EKB-569 (N-[4-[(3-chloro-4-fluorophenyl)amino]-3-cyano-7-ethoxy-6-quinolynyl]-4-(dimethylamino)-2-butynamide) (Wyeth); AG1478 (Pfizer); AG1571 (SU5271, Pfizer); and dual EGFR / HER2 tyrosine kinase inhibitors such as lapatinib (TYKERB (registered trademark), GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2-methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolineamine).

[0180] As chemotherapeutic agents, "tyrosine kinase inhibitors", for example, the EGFR-targeted drugs described in the previous paragraph; inhibitors of insulin receptor tyrosine kinase, for example, inhibitors of anaplastic lymphoma kinase (Alk), such as AF-802 (also known as CH-5424802 or alectinib), ASP3026, X396, LDK378, AP26113, crizotinib (XALKORI (registered trademark)), and ceritinib (ZYKADIA (registered trademark)), etc.; small molecule HER2 tyrosine kinase inhibitors, such as TAK165 available from Takeda; CP-724,714 (Pfizer and OSI), an oral selective inhibitor of ErbB2 receptor tyrosine kinase; dual HER inhibitors, such as EKB-569 (available from Wyeth) that preferentially binds to EGFR but inhibits both HER2 and EGFR overexpressing cells; lapatinib (GSK572016, available from Glaxo-SmithKline); oral HER2 and EGFR tyrosine kinase inhibitors; PKI-166 (available from Novartis); pan-HER inhibitors, such as canertinib (CI-1033, Pharmacia); Raf-1 inhibitors, such as the antisense agent ISIS-5132 available from ISIS Pharmaceuticals that inhibits Raf-1 signaling; non-HER target TK inhibitors, such as imatinib mesylate (GLEEVEC (registered trademark), available from Glaxo SmithKline); multi-target tyrosine kinase inhibitors, such as sunitinib (SUTENT (registered trademark), available from Pfizer); VEGF receptor tyrosine kinase inhibitors, such as bevacizumab (PTK787 / ZK222584, available from Novartis / Schering AG); MAPK extracellular regulatory kinase I inhibitor CI-1040 (available from Pharmacia); quinazolines, such as PD 153035, 4-(3-chloroanilino)quinazoline; pyridopyrimidines; pyrimidopyrimidines; pyrrolopyrimidines, such as CGP 59326, CGP 60261, and CGP 62706; pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine; curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide);Tilphostin containing a nitrothiophene moiety; PD-0183805 (Warner-Lambert); antisense molecules (e.g., those that bind to HER-encoding nucleic acids); quinoxaline (U.S. Patent No. 5,804,396); tryphostin (U.S. Patent No. 5,804,396); ZD6474 (Astra Zeneca); PTK-787 (Novartis / Schering AG); pan-HER inhibitors, e.g., CI-1033 (Pfizer); Affinitac (ISIS 3521; Isis / Lilly); imatinib mesylate (GLEEVEC®); PKI 166 (Novartis); GW2016 (Glaxo SmithKline); CI-1033 (Pfizer); EKB-569 (Wyeth); semaxinib (Pfizer); ZD6474 (AstraZeneca); PTK-787 (Novartis / Schering AG); INC-1C11 (Imclone), rapamycin (sirolimus, RAPAMUNE®); or those described in any of the following patent publications: U.S. Patent No. 5,804,396; International Publication No. WO 1999 / 09016 (American Cyanamid); International Publication No. WO 1998 / 43960 (American Cyanamid); International Publication No. WO 1997 / 38983 (Warner Lambert); International Publication No. WO 1999 / 06378 (Warner Lambert); International Publication No. WO 1999 / 06396 (Warner Lambert); International Publication No. WO 1996 / 30347 (Pfizer, Inc); International Publication No. WO 1996 / 33978 (Zeneca); International Publication No. WO 1996 / 3397 (Zeneca); and International Publication No. WO 1996 / 33980 (Zeneca).;

[0181] Chemotherapeutic agents include dexamethasone, interferon, colchicine, methotrexate, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostine, arsenic trioxide, asparaginase, live BCG, bevacizumab, bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, erlotinib, filgrastim, histrelin acetate, ibritumomab, interferon alpha-2a, interferon alpha-2b, lenalidomide, levamisole, mesna, methoxsalen, nandrolone, nelarabine, nolfetumomab, oprelvekin, palifermin, pamidronate, pegademase, pegasparaginase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium, quinacrine, rasburicase, sargramostim, temozolomide, VM-26, 6-TG, tamoxifen, tretinoin, ATRA, valrubicin, zoledronate, and zoledronic acid, and their pharmaceutically acceptable salts are also included.

[0182] Chemotherapeutic agents include hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivolate, triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, fludrocortolone, hydrocortisone-17-butyrate, hydrocortisone-17-valerate, acrometasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasone-17-propionate, fludrocortolone caproate, fludrocortolone pivolate, and fluprednidene acetate; immune-selective anti-inflammatory peptides (ImSAIDs) such as phenylalanine-glutamine-glycine (FEG) and its D-form (feG) (IMULAN BioTherapeutics, LLC); antirheumatic drugs such as azathioprine, cyclosporine (cyclosporine A), D-penicillamine, gold salts, hydroxychloroquine, leflunomide, minocycline, sulfasalazine; tumor necrosis factor alpha (TNFα) blockers such as etanercept (Enbrel), infliximab (Remicade), adalimumab (Humira), certolizumab pegol (Cimzia), golimumab (Simponi); interleukin 1 (IL-1) blockers such as anakinra (Kineret); T-cell costimulation blockers such as abatacept (Orencia); interleukin 6 (IL-6) blockers such as tocilizumab (ACTEMERA (registered trademark)); interleukin 13 (IL-13) blockers such as lebrikizumab; interferon alpha (IFN) blockers such as lonafarnib; beta7 integrin blockers such as rhuMAb Beta7; IgE pathway blockers such as anti-M1 prime; secretory homotrimeric LTa3 and membrane-bound heterotrimeric LTa1 / β2 blockers such as anti-lymphotoxin alpha (LTa); radioisotopes (e.g., radioisotopes of At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32, Pb212, and Lu);Various investigational drugs such as thioplatin, PS-341, phenylbutyrate, ET-18-OCH3, and farnesyltransferase inhibitors (L-739749, L-744832); polyphenols such as quercetin, resveratrol, piceatannol, epigallocatechin gallate, theaflavin, flavanol, procyanidin, betulinic acid and its derivatives; autophagy inhibitors such as chloroquine; delta-9-tetrahydrocannabinol (dronabinol, MARINOL (registered trademark)); beta-lapachone; lapachol; colchicine; betulinic acid; acetylcamptothecin, scoplectin, and 9-aminocamptothecin); podophyllotoxin; tegafur (UFTORAL (registered trademark)); bexarotene (TARGRETIN (registered trademark)); bisphosphonates such as clodronate (e.g., BONEFOS (registered trademark) or OSTAC (registered trademark)), etidronate (DIDROCAL (registered trademark)), NE-58095, zoledronic acid / zoledronate (ZOMETA (registered trademark)), alendronate (FOSAMAX (registered trademark)), pamidronate (AREDIA (registered trademark)), tiludronate (SKELID (registered trademark)), or risedronate (ACTONEL (registered trademark)); and epidermal growth factor receptor (EGF-R); vaccines such as THERATOPE (registered trademark) vaccine; perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteasome inhibitors (e.g., PS341); CCI-779; tipifarnib (R11577); obatoclax, ABT510; Bcl-2 inhibitors such as oblimersen sodium (GENASENSE (registered trademark)); pixantrone; farnesyltransferase inhibitors such as lonafarnib (SCH 6636, SARASARTM); and pharmaceutically acceptable salts, acids, or derivatives of any of the above; and combinations of two or more of CHOP, an abbreviation for a combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone, and FOLFOX, an abbreviation for a treatment regimen using oxaliplatin (ELOXATIN (trademark)) in combination with 5-FU and leucovorin).;

[0183] Chemotherapeutic agents may also include non-steroidal anti-inflammatory drugs having analgesic, antipyretic, and anti-inflammatory effects. NSAIDs include non-selective inhibitors of the enzyme cyclooxygenase. Specific examples of NSAIDs include propionic acid derivatives such as aspirin, ibuprofen, fenoprofen, ketoprofen, flurbiprofen, oxaprozin, and naproxen; acetic acid derivatives such as indomethacin, sulindac, etodolac, and diclofenac; enolic acid derivatives such as piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, and isoxicam; fenamic acid derivatives such as mefenamic acid, meclofenamic acid, flufenamic acid, and tolfenamic acid; and COX-2 inhibitors such as celecoxib, etoricoxib, lumiracoxib, parecoxib, rofecoxib, rofecoxib, and valdecoxib. NSAIDs may be indicated for the relief of symptoms of conditions such as rheumatoid arthritis, osteoarthritis, inflammatory arthropathy, ankylosing spondylitis, psoriatic arthritis, Reiter's syndrome, acute gout, dysmenorrhea, metastatic bone pain, headache, and migraine, postoperative pain, mild to moderate pain due to inflammation and tissue injury, fever, intestinal obstruction, and renal colic.

[0184] An "effective amount" 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 thereof (e.g., a pharmaceutical composition), is at least the minimum amount necessary to achieve a measurable increase in a desired therapeutic outcome, e.g., the overall survival or progression-free survival of a particular disease or disorder (e.g., cancer, e.g., lung cancer (e.g., NSCLC)). The effective amount herein can vary depending on factors such as the patient's disease state, age, gender, and weight, as well as the ability of the antibody to elicit a desired response in the subject. The effective amount is also one in which the therapeutically beneficial effects exceed any toxic or detrimental effects of the treatment. Beneficial or desired results for prophylactic use include removal or reduction of risk, reduction of severity, or delay in the onset of disease, including biochemical, histological and / or kinetic symptoms of the disease, its complications, and intermediate pathological phenotypes that appear during the development of the disease.For therapeutic use, beneficial or desired results include a reduction in one or more symptoms resulting from a disease (e.g., reduction or delay of cancer-related pain, symptomatic skeletal-related events (SSE), reduction of symptoms by the European Organization for Research and Treatment of Cancer Quality-of-Life Questionnaire (EORTC QLQ-C30, e.g., fatigue, nausea, vomiting, pain, dyspnea, insomnia, anorexia, constipation, diarrhea, or general levels of physical, cognitive, or social function), e.g., reduction of pain measured by a numerical rating scale (NRS) of pain severity of 10 points (measured at worst), and / or reduction of symptoms related to lung cancer by the Symptom in Lung Cancer (SILC) scale (e.g., time to deterioration (TTD) in cough, dyspnea, and chest pain), etc., clinical outcomes such as an increase in the quality of life of a person suffering from the disease, a reduction in the dosage of other drugs required to treat the disease, enhancement of the effect of another drug by targeting, etc., delay in the progression of the disease (e.g., progression-free survival or radiation progression-free survival (rPFS); delay in clear clinical progression (e.g., progression of cancer-related pain, symptomatic skeletal-related events, decrease in Eastern Cooperative Group Oncology Group (ECOG) Performance Status (PS) (e.g., how the disease affects the patient's ability to perform daily activities), and / or initiation of the next systemic anti-cancer therapy), and / or delay in the time to lung-specific antigen progression), and / or extension of survival period. In the case of cancer or tumor, an effective amount of the drug can have the effect of reducing the number of cancer cells, reducing the tumor size, inhibiting the invasion of cancer cells into peripheral organs (i.e., delaying to some extent or preferably stopping), inhibiting tumor metastasis (i.e., delaying to some extent or preferably stopping), inhibiting tumor growth to some extent, and / or reducing to some extent one or more of the symptoms associated with the disorder. The effective amount can be administered in one or more administrations. For the purposes of the present invention, an effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to directly or indirectly achieve a prophylactic treatment or a therapeutic treatment.As understood in the clinical field, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in combination with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" may be considered in the context of administration of one or more therapeutic agents, and a single agent may be considered to be administered in an effective amount in combination with one or more other agents if a desired result can be achieved, or is achieved.

[0185] "Individual response" or "response" can be evaluated using any endpoint indicating benefit to a subject, including, without limitation, (1) some inhibition of disease progression (e.g., progression of cancer, such as lung cancer (e.g., NSCLC)) including deceleration and complete cessation, (2) reduction in tumor size, (3) inhibition (i.e., reduction, deceleration, or complete cessation) of cancer cell invasion into adjacent peripheral organs and / or tissues, (4) inhibition of metastasis (i.e., reduction, deceleration, or complete cessation), (5) some alleviation of one or more symptoms associated with a disease or disorder (e.g., cancer, such as lung cancer (e.g., NSCLC)), (6) increase or prolongation of survival duration including overall survival and progression-free survival, and / or (9) decrease in mortality at a given time point after treatment.

[0186] As used herein, "complete response" or "CR (complete response)" refers to the disappearance of all target lesions.

[0187] As used herein, "partial response" or "PR (partial response)" refers to at least a 30% decrease in the sum of the longest diameters (SLD) of target lesions, with reference to the baseline SLD.

[0188] As used herein, "objective response rate" (ORR) means the sum of the complete response (CR) rate and the partial response (PR) rate.

[0189] The "effective response" of a subject to treatment with a pharmaceutical, or the "responsiveness" of a subject, and similar terms, refer to the clinical or therapeutic benefit conferred on a subject at risk of or suffering from a disease or disorder such as cancer. In one embodiment, such benefits include any one or more of: extending the survival period (including overall survival and progression-free survival), resulting in an objective response (including complete response or partial response), or improving the signs or symptoms of cancer.

[0190] A subject who "does not show an effective response" to treatment refers to a subject who does not have any of: an extended survival period (including overall survival and progression-free survival), an objective response (including complete response or partial response), or improved signs or symptoms of cancer.

[0191] As used herein, the term "survive" refers to a patient being alive, including overall survival and progression-free survival.

[0192] As used herein, "overall survival" (OS) refers to the proportion of subjects in a group who are alive at a specific time period, for example, one year or five years after diagnosis or treatment.

[0193] As used herein, "progression-free survival" (PFS) refers to the length of time during and after treatment during which the treated disease (e.g., cancer, such as lung cancer (e.g., NSCLC)) does not progress. Progression-free survival can include the amount of time a patient experiences a complete response or partial response, as well as the amount of time a patient experiences stable disease.

[0194] As used herein, "stable disease" or "SD" refers to the absence of sufficient shrinkage of target lesions to be called a PR or sufficient increase to be called a PD, based on the minimum SLD since the start of treatment.

[0195] As used herein, "progressive disease" or "PD" refers to an increase of at least 20% in the SLD of a target lesion, based on the minimum SLD recorded after the start of treatment or after the presence of one or more new lesions.

[0196] As used herein, "retarding the progression of" a disease or disorder refers to delaying, preventing, retarding, slowing, stabilizing, and / or postponing the development of a disease or disorder (e.g., cancer, such as lung cancer (e.g., NSCLC)). This delay can be of various durations depending on the medical history and / or the subject being treated. As will be apparent to those skilled in the art, a sufficient or significant delay can effectively encompass prevention in that the subject does not develop the disease. For example, in advanced cancer, the development of central nervous system (CNS) metastases can be delayed.

[0197] By "survival prolongation" is meant an increase in the overall survival or progression-free survival in a treated patient relative to an untreated patient (e.g., a patient not treated with a pharmaceutical), or a patient not expressing a biomarker at a specified level, and / or a patient treated with an approved anti-tumor agent. Objective response refers to a measurable response that includes a complete response (CR) or a partial response (PR).

[0198] As used herein, "hazard ratio" or "HR" is a statistical definition of the event incidence rate. For the purposes of the present 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 at any given point in time. An HR value of 1 indicates that the relative risk of the endpoint (e.g., death) is equal in both the "treatment" and "control" groups, a value greater than 1 indicates a higher risk in the treatment group compared to the control group, and a value less than 1 indicates a higher risk in the control group compared to the treatment group. The "hazard ratio" (i.e., PFS HR) in progression-free survival analysis is the sum of the differences between two progression-free survival curves and represents the reduction in the risk of death in the treatment compared to the control over the follow-up period. The "hazard ratio" (i.e., OS HR) in overall survival analysis is the sum of the differences between two overall survival curves and represents the reduction in the risk of death in the treatment compared to the control over the follow-up period.

[0199] As used herein, the "Ventana SP263 IHC assay" (also referred to herein as the Ventana SP263 CDx assay) is performed according to the Ventana PD-L1 (SP263) assay package insert (Tucson, AZ: Ventana Medical Systems, Inc.), which is hereby incorporated by reference in its entirety.

[0200] As used herein, the "Ventana SP142 IHC assay" is performed according to the Ventana PD-L1 (SP142) assay package insert (Tucson, AZ: Ventana Medical Systems, Inc.), which is hereby incorporated by reference in its entirety.

[0201] As used herein, the “pharmDx 22C3 IHC assay” is performed in accordance with the PD-L1 IHC 22C3 pharmDx package insert (Carpinteria, CA: Dako, Agilent Pathology Solutions), which is hereby incorporated by reference in its entirety.

[0202] As used herein, the term “tumor-infiltrating immune cells” refers to any immune cells present in a tumor or a sample thereof. Tumor-infiltrating immune cells include, but are not limited to, immune cells within the tumor, immune cells surrounding the tumor, 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 granulocytes (e.g., neutrophils, eosinophils, and basophils), monocytes, macrophages, dendritic cells (e.g., finger-like interdigitating dendritic cells), histiocytes, and other myeloid cells including natural killer cells.

[0203] As used herein, the term “biomarker” refers to an indicator that can be detected in a sample, such as a predictive indicator, a diagnostic indicator, and / or a prognostic indicator. In some embodiments, the biomarker is a gene. Biomarkers include, but are not limited to, polypeptides, polynucleotides (e.g., DNA and / or RNA), changes in polynucleotide copy number (e.g., DNA copy number), polypeptide and polynucleotide modifications (e.g., post-translational modifications), carbohydrates, and / or glycolipid-based molecular markers.

[0204] The term “antibody” includes monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region), antibody compositions having polyepitope specificity, multispecific antibodies (e.g., bispecific antibodies, diabodies, 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 herein synonymously with “antibody”.

[0205] The basic four-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light chains (L) and two identical heavy chains (H). IgM antibodies consist of five basic heterotetrameric units together with an additional polypeptide called the J chain and contain ten antigen-binding sites, while IgA antibodies are composed of two to five basic four-chain units, which can polymerize to form multivalent aggregates in combination with the J chain. In the case of IgG, the four-chain unit generally weighs about 150,000 daltons. Each L chain is linked to the H chain by one disulfide covalent bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. The H and L chains each also have regularly spaced interchain disulfide bridges. Each H chain has a variable domain (V H ) at the N-terminus, followed by three constant domains (C H ) for each of the α and γ chains, and four C H domains for the μ and ε isotypes. Each L chain has a variable domain (V L ) at the N-terminus and a constant domain at the opposite end. V L is aligned with V H , and C L is aligned with the first constant domain (C H 1) of the heavy chain. Certain amino acid residues are thought to form an interface between the light chain variable domain and the heavy chain variable domain. V H and V LBy pairing together, a single antigen-binding site is formed. For the structures and properties of various classes of antibodies, see, for example, pages 71 and Chapter 6 of Basic and Clinical Immunology, 8th Edition, Daniel P. Sties, Abba I. Terr and Tristram G. Parsolw (eds), Appleton & Lange, Norwalk, CT, 1994. Light chains derived from any vertebrate species can be assigned to one of two distinct types called kappa and lambda based on the amino acid sequences of their constant domains. Depending on the amino acid sequences of their heavy chain (CH) constant domains, immunoglobulins can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, each having heavy chains designated as α, δ, ε, γ, and μ, respectively. The γ and α classes are further classified into subclasses based on relatively minor differences in CH sequences and functions. For example, humans express the following subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1, and IgA2.

[0206] As used herein, the terms "hypervariable region" or "HVR" refer to each region of an antibody variable domain that is hypervariable in sequence and determines antigen-binding specificity, e.g., each of the "complementary determining regions" (CDRs).

[0207] Generally, an antibody contains six CDRs, three in VH (CDR-H1, CDR-H2, CDR-H3) and three in VL (CDR-L1, CDR-L2, CDR-L3). Exemplary CDRs herein include (a) hypervariable loops occurring at amino acid residues 26 - 32 (L1), 50 - 52 (L2), 91 - 96 (L3), 26 - 32 (H1), 53 - 55 (H2), and 96 - 101 (H3) (Chothia and Lesk, J. Mol. Biol. 196:901 - 917 (1987)), (b) CDRs present in 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 (c) antigen contacts occurring at amino acid residues 27c - 36 (L1), 46 - 55 (L2), 89 - 96 (L3), 30 - 35b (H1), 47 - 58 (H2), and 93 - 101 (H3) (MacCallum et al. J. Mol. Biol. 262:732 - 745 (1996)).

[0208] Unless otherwise specified, CDRs are determined according to Kabat et al. above. One of ordinary skill in the art will understand that the naming of CDRs can also be determined according to Chothia et al. above, McCallum et al. above, or any other scientifically recognized nomenclature.

[0209] The expressions "variable domain residue numbering as in Kabat" or "amino acid position numbering as in Kabat", and variations thereof, refer to the numbering scheme used for the heavy - chain variable domain or the light - chain variable domain in the antibody compilation in Kabat et al. above. Using this numbering scheme, the actual linear amino acid sequence can include fewer or additional amino acids corresponding to deletions or insertions in the FR or HVR of the variable domain. For example, the heavy - chain variable domain can include a single amino acid insertion (residue 52a according to Kabat) after residue 52 of H2, and residues inserted after residue 82 of the heavy - chain FR (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). The Kabat numbering of residues can be determined for a given antibody by alignment in the homologous regions between the sequence of the antibody and the sequence numbered by "standard" Kabat.

[0210] The term "variable" refers to the fact that certain segments of the variable domains vary widely between antibodies in an array. The V domains mediate antigen binding and define the specificity of a particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domain. Rather, it is concentrated in three segments called hypervariable regions (HVRs) in the variable domains of both the light and heavy chains. The more highly conserved portions of the variable domains are called framework regions (FRs). The variable domains of native heavy and light chains each adopt a beta-sheet conformation connected by three HVRs that form loops connecting, and in some cases forming part of, the beta-sheet structure, and contain four FR regions that largely adopt a beta-sheet conformation. The HVRs within each chain are brought into proximity and linked to each other by the FR regions and, together with the HVRs of the other chain, contribute to the formation of the antigen-binding site of the antibody (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, MD (1991)). The constant domains do not directly participate in the binding of the antibody to the antigen but exhibit various effector functions such as the involvement of the antibody in antibody-dependent cell cytotoxicity.

[0211] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of the heavy or light chain of the antibody. The variable domains of the heavy and light chains may be referred to as "VH" and "VL", respectively. These domains are generally the most variable parts of the antibody (relative to other antibodies of the same class) and contain the antigen-binding site.

[0212] "Framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FRs of the variable domain generally consist of four FR domains: the FR1 domain, the FR2 domain, the FR3 domain, and the FR4 domain. Thus, the HVR and FR sequences generally appear in the following sequence in VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0213] The terms "full-length antibody", "intact antibody", and "whole antibody" are used interchangeably to refer to an antibody in its substantially intact form, in contrast to antibody fragments. Specifically, whole antibodies include those having heavy and light chains that include an Fc region. The constant domains can be the constant domains of the native sequence (e.g., the constant domains of the human native sequence) or amino acid sequence variants thereof. In some cases, intact antibodies can have one or more effector functions.

[0214] "Antibody fragment" includes a portion of an intact antibody, preferably the antigen-binding region and / or variable region of an intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab’, F(ab’)2; diabodies; linear antibodies (see, e.g., Example 2 of U.S. Patent No. 5,641,870; Zapata et al., Protein Eng. 8(10):1057-1062

[1995] ); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. Papain digestion of an antibody yields two identical antigen-binding fragments termed "Fab" fragments and a residual "Fc" fragment so-called because of its ability to crystallize readily. The Fab fragment consists of the variable region domain (V H ) of the H chain together with the entire L chain, as well as the first constant domain (C HIt consists of 1). Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen-binding site. By pepsin treatment of an antibody, a single large F(ab’)2 fragment is obtained, which is approximately equivalent to two Fab fragments with different antigen-binding activities disulfide-bonded together and can still crosslink to an antigen. The Fab’ fragment differs from the Fab fragment in having several additional residues at the carboxy terminus of the C H 1 domain. Fab’-SH is the name in this specification for a Fab’ in which the cysteine residue(s) of the constant domain has a free thiol group. The F(ab’)2 antibody fragment was originally produced as a pair of Fab’ fragments with a hinge cysteine in between. Other chemical couplings of antibody fragments are also known.

[0215] The Fc fragment contains the carboxy termini of both H chains held together by disulfides. The effector function of an antibody is determined by the sequence in the Fc region, which is also recognized by Fc receptors (FcRs) found on certain cell types.

[0216] The “functional fragment” of the antibody of the present invention includes a portion of the intact antibody, which generally includes the antigen-binding region or variable region of the intact antibody, or the Fc region of an antibody that retains or has a modified FcR-binding ability. Examples of antibody fragments include linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0217] "Fv" is the smallest antibody fragment that contains a complete antigen recognition and antigen-binding site. This fragment consists of a dimer in which one heavy-chain variable region domain and one light-chain variable region domain are tightly bound non-covalently. The folding of these two domains gives rise to six hypervariable loops (three loops each from the H chain and the L chain) that provide amino acid residues for antigen binding and confer antigen-binding specificity on the antibody. However, even a single variable domain (or half of an Fv that contains only three HVRs specific for an antigen) has the ability to recognize and bind the antigen, although with a lower affinity than the full binding site.

[0218] "Single-chain Fv", also abbreviated as "sFv" or "scFv", is an antibody fragment that contains the V H and V L antibody domains joined to form a single polypeptide chain. Preferably, the sFv polypeptide further contains a polypeptide linker between the V H domain and the V L domain, which enables the sFv to assume a structure desirable for antigen binding. For a review of sFv, see, for example, Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenberg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0219] The term "Fc region" as used herein is used to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain may vary, the Fc region of a human IgG heavy chain is typically defined as extending from the amino acid residue at position Cys226 or the amino acid residue at position Pro230 to its carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) may be removed, for example, during the production or purification of an antibody or by recombinant engineering of the nucleic acid encoding the heavy chain of the antibody. Thus, a composition of intact antibodies may include a population of antibodies in which all K447 residues have been removed, a population of antibodies without removed K447 residues, and a population of antibodies having a mixture of antibodies with and without K447 residues. Native sequence Fc regions suitable for use in the antibodies of the present invention include human IgG1, IgG2 (IgG2A, IgG2B), IgG3, and IgG4. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region follows the EU numbering system, also called the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0220] "Fc receptor" or "FcR" refers to a receptor that binds to the Fc region of an antibody. Preferred FcRs are native sequence human FcRs. Further, preferred FcRs are those that bind to IgG antibodies (gamma receptors), including receptors of the FcγRI, FcγRII, and FcγRIII subclasses (including allelic variants and alternatively spliced forms of these receptors). Among the FcγRII receptors are FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences that mainly differ in their cytoplasmic domains. The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) within its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic domain (see M. Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein.

[0221] The term "diabody" refers to a small antibody fragment prepared by constructing an sFv fragment (see the previous paragraph) using a short linker (about 5-10 residues) between the V H domain and the V L domain to achieve inter-chain rather than intra-chain V domain pairing, thereby obtaining a bivalent fragment, i.e., a fragment having two antigen-binding sites. A bispecific diabody is composed of the V H domains and the V LIt is a heterodimer of two "crossed" sFv fragments where the domains are present in different polypeptide chains. Diabodies are described in detail, for example, in European Patent No. 404,097, International Publication No. 93 / 11161; Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993).

[0222] As monoclonal antibodies herein, specifically, a "chimeric" antibody (immunoglobulin) in which a portion of the heavy chain and / or light chain is identical or homologous to the corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, and fragments of such antibodies as long as they exhibit the desired biological activity (U.S. Patent No. 4,816,567, Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). Chimeric antibodies for the purposes herein include PRIMATIZED™ antibodies, where the antigen-binding region of this antibody is derived from an antibody produced, for example, by immunizing a cynomolgus macaque with the antigen of interest. As used herein, "humanized antibody" is used as a subset of "chimeric antibody".

[0223] The "class" of an antibody refers to the type of constant domain or constant region carried by its heavy chain. There are five main classes of antibodies, namely, IgA, IgD, IgE, IgG, and IgM, and some of these may be further divided into subclasses (isotypes), for example, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0224] "Affinity" refers to the strength of the total non-covalent interactions between a single binding site of a molecule (e.g., an antibody, e.g., TIGIT or PD-L1) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects the 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y can generally be represented by the dissociation constant (K D ). Affinity can be measured by methods common in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0225] "Human antibody" refers to an antibody having an amino acid sequence corresponding to the amino acid sequence of an antibody produced by a human, and / or an antibody produced using any of the techniques for producing human antibodies disclosed herein. This definition of human antibody clearly excludes humanized antibodies containing non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, such as phage display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). The methods 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) can also be utilized for the preparation of human monoclonal antibodies. 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, such as an immunized xenomouse, that has been modified to produce such antibodies in response to antigen administration but whose endogenous locus is rendered inactive (see, for example, U.S. Patent Nos. 6,075,181 and 6,150,584 regarding XENOMOUSE™ technology). For human antibodies produced by human B cell hybridoma technology, see, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006).

[0226] A "humanized" non-human (e.g., murine) antibody is a chimeric antibody that contains minimal sequences derived from non-human immunoglobulins. In one embodiment, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's HVRs (defined below) have been replaced with residues from the HVRs of a non-human species such as a mouse, rat, rabbit, or non-human primate (donor antibody) that have the desired specificity, affinity, and / or activity. In some instances, framework ("FR") residues of the human immunoglobulin are replaced with the corresponding non-human residues. Additionally, a humanized antibody may contain residues not found in the recipient antibody or donor antibody. These modifications can be made to further improve antibody performance, such as binding affinity. Generally, a humanized antibody is one that includes substantially all of at least one, typically two, variable domains, with all or substantially all of the hypervariable loops corresponding to those of a non-human immunoglobulin sequence and all or substantially all of the FR regions corresponding to those of a human immunoglobulin sequence, although the FR regions may include one or more individual FR residue substitutions that improve antibody performance, such as binding affinity, isomerization, immunogenicity, etc. The number of these amino acid substitutions in the FRs is typically 6 or less for the H chain and 3 or less for the L chain. A humanized antibody optionally also includes the immunoglobulin constant region (Fc), typically at least a portion of the Fc of a human immunoglobulin. For further details, see, e.g., 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, e.g., 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.

[0227] The term "isolated antibody" as used herein to describe the various antibodies disclosed herein means an antibody that has been identified, separated and / or recovered from the cells or cell culture in which it was expressed. Contaminating components of its natural environment are materials that typically interfere with the diagnostic or therapeutic use of the polypeptide and can include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In some embodiments, the antibody is purified to greater than 95% or greater than 99% purity as determined, for example, by electrophoresis (e.g., SDS-PAGE, isoelectric focusing electrophoresis (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse 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 is purified to homogeneity by SDS-PAGE under non-reducing or reducing conditions using (1) a spinning cup sequenator to obtain at least 15 residues of the N-terminal or internal amino acid sequence to a sufficient extent, or (2) Coomassie blue or preferably silver staining. Isolated antibodies include antibodies in situ within recombinant cells, since at least one component of the polypeptide's natural environment is absent. However, usually, an isolated polypeptide is prepared by at least one purification step.

[0228] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation) that may be present in minor amounts. Monoclonal antibodies are highly specific and are directed against a single antigenic site. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they are synthesized by hybridoma culture and are free of other immunoglobulins. The modifier "monoclonal" indicates the characteristic of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies used in accordance with the present invention may be obtained by, for example, the hybridoma method (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, 2 nd(ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567), phage display techniques (see, e.g., 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 or human-like antibodies in animals having a part or all of the human immunoglobulin locus or a gene encoding a human immunoglobulin sequence (see, e.g., International Publication No. WO 1998 / 24893; International Publication No. WO 1996 / 34096; International Publication No. WO 1996 / 33735; International Publication No. 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); U.S. Patent No. 5,545,807; No. 5,545,806; No. 5,569,825; No. 5,625,126; No. 5,633,425; and No. 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.It can be prepared by various techniques including, see, e.g., 14:845-851(1996); Neuberger, Nature Biotechnol. 14:826(1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13:65-93(1995).

[0229] The "percent amino acid sequence identity (%)" with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues of the reference polypeptide, after aligning the sequences and introducing gaps if necessary to obtain the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignments for the purpose of determining percent amino acid sequence identity can be achieved in a variety of ways within the scope of techniques in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. One of ordinary skill in the art can determine appropriate parameters for aligning the sequences, including any algorithm necessary to achieve the maximum alignment with respect to the full length of the sequences being compared. However, for the purposes herein, the percent amino acid sequence identity values are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was created by Genentech, Inc., and the source code was submitted to the U.S. Copyright Office, Washington D.C., 20559, together with user documentation, and is registered therein as U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc. (South San Francisco, California) or can be compiled from its source code. The ALIGN-2 program should be compiled for use in a UNIX operating system, including Digital UNIX V4.0D. All sequence comparison parameters are set by and are not changed by the ALIGN-2 program.

[0230] In the situation where ALIGN-2 is used for amino acid sequence comparison, the amino acid sequence identity % of a given amino acid sequence A to, with, or against a given amino acid sequence B (alternatively, can be described as a given amino acid sequence A having or including a specific amino acid sequence identity % to, with, or against a given amino acid sequence B) is calculated as follows. 100 × fraction X / Y

[0231] In this case, X is the number of amino acid residues scored as identical matches in the alignment of A and B by the sequence alignment program ALIGN-2, and Y is the total number of amino acid residues in B. It will be understood that if the length of amino acid sequence A is different from the length of amino acid sequence B, the % amino acid sequence identity of A to B will be different from the % amino acid sequence identity of B to A. Unless otherwise specified, when used in this specification, all % amino acid sequence identity values are obtained using the ALIGN-2 computer program as described in the previous paragraph.

[0232] As used herein, "subject" or "individual" means a mammal, including but not limited to human or non-human mammals such as cows, horses, dogs, sheep, or cats. In some embodiments, the subject is a human. A patient is also a subject as used herein.

[0233] As used herein, the term "sample" refers to a composition obtained from or derived from a target subject and / or individual that contains entities of cells and / or other molecules that are characterized and / or identified, for example, based on physical, biochemical, chemical, and / or physiological properties. For example, the phrases "tumor sample", "disease sample" and variations thereof refer to any sample obtained from a subject of interest that is expected to contain or is known to contain the characterized cell and / or molecular entity. 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 humor, lymph, synovial fluid, follicular fluid, semen, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, feces, tumor lysates, and tissue culture media, tissue extracts, e.g., homogenized tissue, cell extracts, and combinations thereof.

[0234] The terms "tissue sample" and "cell sample" mean a collection of similar cells obtained from the tissue of a subject or individual. The source of the tissue or cell sample can be solid tissue such as fresh, frozen, and / or preserved organs, tissue samples, biopsy materials, and / or aspirates; blood or any blood component such as plasma; body fluids such as cerebrospinal fluid, amniotic fluid, ascites, or interstitial fluid; cells at any stage of the subject's pregnancy or development. A tissue sample can also be a primary cell or a cultured cell or cell line. Optionally, the tissue or cell sample is obtained from diseased tissue / organs. The tissue sample can contain compounds that do not naturally mix with natural tissue, such as preservatives, anticoagulants, buffers, fixatives, nutrients, or antibiotics.

[0235] 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 comparison purposes. In one embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and / or non-affected part (e.g., tissue or cell) of the body of the same subject. For example, healthy and / or non-affected cells or tissues adjacent to the diseased cells or tissues (e.g., cells or tissues adjacent to a tumor). In another embodiment, the reference sample is obtained from untreated tissue and / or cells of the body 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 non-affected part (e.g., tissue or cell) of the body of a non-subject. In yet another embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from untreated tissue and / or cells of the body of an individual who is not the subject.

[0236] Unless otherwise specified, as used herein, the term "protein" refers to any native protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice, rats), unless otherwise specified. This term includes any form of the "full-length", untreated protein and the protein resulting from processing within the cell. This term also encompasses naturally occurring variants of the protein, such as splice variants or allelic variants.

[0237] As used interchangeably herein, "polynucleotide" or "nucleic acid" refers to a polymer of nucleotides of any length and includes DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthetic reaction. Thus, for example, polynucleotides as defined herein include single-stranded and double-stranded DNA, DNA containing single-stranded and double-stranded regions, single-stranded and double-stranded RNA, and RNA containing single-stranded and double-stranded regions, hybrid molecules comprising DNA and RNA that contain single-stranded or more typically double-stranded regions or that can contain single-stranded and double-stranded regions, but are not limited thereto. In addition, as used herein, the term "polynucleotide" refers to triple-stranded regions containing RNA or DNA, or both RNA and DNA. The strands within such regions can be from the same molecule or from different molecules. These regions can include all of one or more of these molecules, but more typically include only some regions of some of these molecules. One of the molecules in the triple helix region is often an oligonucleotide. The terms "polynucleotide" and "nucleic acid" specifically include mRNA and cDNA.

[0238] The polynucleotide may contain modified nucleotides such as methylated nucleotides and their analogs. If present, the modifications to the nucleotide structure may be imparted either before or after the assembly of the polymer. The nucleotide sequence may be interrupted by non-nucleotide components. The polynucleotide may be further modified after synthesis, such as by conjugation with a label. Other types of modifications include substitution with one or more analogs of naturally occurring nucleotides, internucleotide modifications, for example, by non-charged linkages (e.g., methylphosphonates, phosphotriesters, phosphoramidates, carbamates, etc.) and charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), pendant moieties, such as those containing proteins (e.g., nucleases, toxins, antibodies, signal peptides, poly-L-lysine, etc.), those by intercalating agents (e.g., acridines, psoralens, etc.), those containing chelating agents (e.g., metals, radioactive metals, boron, metal oxides, etc.), those containing alkylating agents, those by modified linkages (e.g., alpha-anomer nucleic acids, etc.), as well as the unmodified form(s) of the polynucleotide(s). Further, any of the hydroxyl groups normally present in the sugar may be replaced, for example, by phosphonate or phosphate groups, protected by standard protecting groups, or activated to prepare for an additional linkage to an additional nucleotide, or conjugated to a solid or semi-solid support. The 5' and 3' terminal OHs may be phosphorylated or substituted with an amine or an organic capping group moiety of 1 to 20 carbon atoms. Other hydroxyls may also be derivatized with standard protecting groups. The polynucleotide may also include similar forms of ribose or deoxyribose sugars commonly known in the art, such as, for example, 2'-O-methyl-, 2'-O-allyl-, 2'-fluoro-, or 2'-azido-ribose, analogs of carbocyclic sugars, alpha-anomer sugars, epimeric sugars, such as arabinose, xylose, or lyxose, pyranose sugars, furanose sugars, sedoheptulose, acrylic acid analogs, and abasic nucleoside analogs, such as methyl riboside.One or more phosphodiester linkages may be replaced by alternative linking groups. These alternative linking groups include embodiments in which the phosphate is replaced by P(O)S (“thioate”), P(S)S (“dithioate”), (O)NR2 (“amidate”), P(O)R, P(O)OR’, CO, or CH2 (“formacetal”), but are not limited thereto, where each R or R’ is independently H or substituted or unsubstituted alkyl (1-20C) (optionally including an ether (-O-) bond), aryl, alkenyl, cycloalkyl, cycloalkenyl, or araldyl. Not all of the linkages in the polynucleotide need to be the same. The foregoing description applies to all polynucleotides referred to herein, including RNA and DNA.

[0239] As used herein, “carrier” includes a pharmaceutically acceptable carrier, excipient, or stabilizer that is nontoxic to the cells or mammals being exposed thereto at the dosages and concentrations employed. In many cases, the physiologically acceptable carrier is an aqueous, pH buffered solution. Examples of physiologically acceptable carriers include buffers such as phosphoric acid, citric acid, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.

[0240] The phrase “pharmaceutically acceptable” indicates that the substance or composition must be chemically and / or toxicologically compatible with the other components of the formulation and / or the mammal being treated therewith.

[0241] The term "pharmaceutical preparation" refers to a preparation in a form such that the biological activity of the active ingredient contained in the preparation is effective and that does not contain further constituents that are unacceptably toxic to the subject to which the preparation is administered.

[0242] III. Prognostic Diagnostic Methods and Assays A. Tumor-Associated Macrophage (TAM) Genes and Gene Signatures A method of identifying an individual who may benefit from treatment (i) TAM genes In one aspect, the present invention is a method of identifying an individual having cancer (e.g., lung cancer, e.g., non-small cell lung cancer (NSCLC)) who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV of this specification and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV of this specification), the method comprising detecting, in a sample from the individual, the expression level of one or more of the tumor-associated macrophage (TAM) genes complement C1q subcomponent C (C1QC), macrophage scavenger receptor type I and type II (MSR1), macrophage mannose receptor 1 (MRC1), V-set and immunoglobulin domain-containing protein 4 (VSIG4), secreted phosphoprotein 1 (SPP1), and macrophage receptor with collagenous structure (MARCO) (e.g., one, two, three, four, five, or all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO), wherein an individual having an expression level of C1QC, MSR1, MRC1, VSIG4, SPP1, or MARCO that exceeds the respective reference expression level is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0243] (ii) TAM signature score In another aspect, the present invention provides a method of identifying an individual having cancer (e.g., lung cancer, e.g., NSCLC) who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV of this specification and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV of this specification), 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 therefrom a tumor-associated macrophage (TAM) signature score, wherein an individual having a TAM signature score above a reference TAM signature score is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0244] In another aspect, the present invention provides a method of identifying an individual having cancer (e.g., lung cancer, e.g., NSCLC) who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV of this specification and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV of this specification), 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 therefrom a TAM signature score, wherein a TAM signature score above a reference TAM signature score identifies the individual as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0245] In some embodiments, the individual has a TAM signature score in a sample that exceeds 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. Exemplary methods for determining the TAM signature score and an exemplary reference TAM signature score are provided below. Exemplary PD-1 axis-binding antagonists, anti-TIGIT antagonist antibodies, and treatment methods comprising these agents are provided in Section IV.

[0246] In some embodiments of any of the methods provided herein, the cancer is lung cancer, such as NSCLC, small cell lung cancer (SCLC), or a lung carcinoid tumor. In some embodiments, the individual is human.

[0247] Method for selecting a treatment (i) TAM gene In another aspect, the invention is a method for selecting a treatment for an individual having cancer (e.g., lung cancer, such as NSCLC), the method comprising detecting the expression level of one or more (e.g., one, two, three, four, five, or all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO) of the tumor-associated macrophage (TAM) genes C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from the individual, and an individual in which the expression level of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO that exceeds their respective reference expression levels is identified as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV of this specification and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV of this specification).

[0248] (ii) TAM signature score In another aspect, the present invention provides a method for selecting a treatment for an individual having cancer (e.g., lung cancer, e.g., 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 signature score therefrom, wherein an individual having a TAM signature score above a reference TAM signature score is identified as an individual likely to benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0249] In another aspect, the present invention provides a method for selecting a treatment for an individual having cancer (e.g., lung cancer, e.g., 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 signature score therefrom, wherein an individual having a TAM signature score above a reference TAM signature score is identified as an individual likely to benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0250] In some aspects, the individual has a TAM signature score in a sample above the 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.

[0251] Treatment method (i) TAM gene In another aspect, the present invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), the method comprising: (a) detecting the expression level of one or more (e.g., one, two, three, four, five, or all six) of the tumor-associated macrophage (TAM) genes C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from the individual, wherein the expression level of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO exceeds the respective reference expression level, thereby identifying the individual as an individual who may benefit from treatment with 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., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0252] In another aspect, the present invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein), wherein the individual has been determined to have an expression level of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO that exceeds the respective reference expression level, thereby identifying the individual as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0253] (ii) TAM signature score In another aspect, the present invention is a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), 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 therefrom a TAM signature score, wherein the TAM signature score exceeds a reference TAM signature score, whereby the individual is identified as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, detecting the expression levels and determining therefrom the TAM signature score; and (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0254] In another aspect, the present invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), 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 exceeds a reference TAM signature score, thereby identifying the individual as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, 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; and (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tirigolumab) disclosed in Section IV herein).

[0255] In another aspect, the present invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tirigolumab) disclosed in Section IV herein), wherein the individual has been determined to have a TAM signature score that exceeds a reference TAM signature score, thereby identifying the individual as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and the TAM signature score is based on the expression levels of at least two (e.g., two, three, four, five, or all six) of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO detected in a sample from the individual.

[0256] In another aspect, the present invention is a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein), wherein the individual has been determined to have a TAM signature score that exceeds a reference TAM signature score, thereby identifying the individual as an individual who may benefit from treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, and 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.

[0257] Advantages Individuals who benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody may experience, for example, a delay or prevention of the development or recurrence of cancer (e.g., lung cancer, e.g., NSCLC), a remission of cancer symptoms, a reduction in any direct or indirect pathological consequences of cancer, a prevention of metastasis, a decrease in the rate of disease progression, an improvement or remission of the disease state, or an improvement in remission or prognosis.

[0258] In some aspects, the benefits achieved by treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody are an increase in progression-free survival (PFS) (e.g., an increase in the duration of PFS experienced by an individual treated according to the method or an increase in the average PFS of a population of individuals treated according to the method), an increase in objective response rate (ORR) (e.g., an increase in the ORR of a population of individuals treated according to the method), and / or an increase in overall survival (OS) (e.g., an increase in the duration of OS experienced by an individual treated according to the method or an increase in the average OS of a population of individuals treated according to the method).

[0259] An increased PFS, ORR, and / or OS can be determined, for example, by comparison to untreated reference individuals and / or a reference population of individuals, a control treatment, such as one or more previously approved treatments or reference individuals and / or a reference population of individuals who received a commercial product for the treatment of cancer; and / or reference individuals and / or a reference population of individuals treated with a PD-1 axis-binding antagonist (e.g., atezolizumab) or an anti-TIGIT antagonist antibody (e.g., tiragolumab) as monotherapy. In some embodiments, the increased PFS, ORR, and / or OS is determined by comparison to reference individuals and / or a reference population of individuals having cancer treated with a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., atezolizumab and tiragolumab), wherein each individual in the reference individuals and / or reference population has a TAM signature score below a reference TAM signature score and / or an expression level of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO below their respective reference expression levels. The reference TAM signature score is described herein and can be, for example, the median TAM signature score of a reference population of individuals having cancer (e.g., lung cancer, e.g., NSCLC) or the median expression level of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a reference population of individuals having cancer (e.g., lung cancer, e.g., NSCLC).

[0260] One of ordinary skill in the art can readily determine whether a given clinical outcome is improved according to the present invention. For example, in this context, "improved" means having an expression level of one or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO that exceeds each respective baseline expression level, or that the clinical outcome resulting from the treatment of an individual having a TAM signature score that exceeds the baseline TAM signature score by treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., atezolizumab and tiragolumab) 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 compared to the clinical outcome resulting from the above-mentioned comparator treatment.

[0261] For example, in some embodiments, the duration of PFS or OS experienced by an individual 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%.

[0262] In another example, in some embodiments, the ORR 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%.

[0263] The time at which the clinical outcome / clinical endpoint is evaluated can be readily determined by one of ordinary skill in the art. In principle, it is determined when the difference in the clinical outcome / clinical endpoint between 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.

[0264] Sample The expression level and / or TAM signature score 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, tumor samples, whole blood samples, plasma samples, serum samples, and combinations thereof. The sample may be fresh, stored, or frozen.

[0265] In some embodiments, the sample is a tissue sample, such as a tumor tissue sample. In some embodiments, the tumor tissue sample is a biopsy material. In some embodiments where the cancer is lung cancer (e.g., NSCLC), the sample is a biopsy material of lung cancer.

[0266] In some embodiments, the sample is obtained from an individual prior to treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, e.g., immediately prior to the first administration of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, or at least 1 day, at least 1 week, or at least 1 month before the first administration of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

[0267] TAM signature score In some embodiments, determining a TAM signature score in a sample from an individual comprises calculating an average of the expression levels of at least two of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual. Thus, in some embodiments, the TAM signature score is an average of the expression levels (e.g., the average of the normalized expression levels) of at least two of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual.

[0268] In some embodiments, determining a TAM signature score in a sample from an individual comprises calculating an average of the expression levels of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual. Thus, in some embodiments, the TAM signature score is an average of the expression levels (e.g., the average of the normalized expression levels) of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in the sample from the individual.

[0269] Expression level The expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, and / or MARCO detected by the methods provided herein can be, for example, nucleic acid expression levels or protein expression levels.

[0270] In some embodiments, the expression level is a nucleic acid expression level, such as an mRNA expression level. The nucleic acid expression level can be detected using any suitable method known in the art, for example, by RNA-seq, reverse transcriptase quantitative 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 a combination thereof. Other amplification-based methods include, for example, transcription-mediated amplification (TMA), strand displacement amplification (SDA), nucleic acid sequence-based amplification (NASBA), and signal amplification methods such as bDNA.

[0271] In some examples, the nucleic acid expression levels of the genes described herein can be measured by sequencing-based techniques such as, for example, RNA-seq, serial analysis of gene expression (SAGE), high-throughput sequencing technology (e.g., massively parallel sequencing), and Sequenom MassARRAY® technology. The nucleic acid expression levels can also be measured, for example, by NanoString nCounter and high coverage expression profiling (HiCEP). Additional protocols for assessing the status of genes and gene products can be found, for example, in Ausubel et al., eds., 1995, Current Protocols In Molecular Biology, Part 2 (Northern blotting), Part 4 (Southern blotting), Part 15 (immunoblotting), and Part 18 (PCR analysis).

[0272] Other methods for detecting the nucleic acid levels of the genes described herein include protocols for examining or detecting mRNAs such as target mRNAs in tissue or cell samples by microarray technology.

[0273] Other methods for detecting the nucleic acid expression level of the genes described in this specification include electrophoresis, Northern blot analysis, and Southern blot analysis, in situ hybridization (e.g., single or multiplex nucleic acid in situ hybridization), RNase protection assay, and microarray (e.g., Illumina BEADARRAY™ technology; bead array for detecting gene expression (BADGE)).

[0274] In some embodiments, the expression level is the protein expression level, e.g., the protein expression level determined by mass spectrometry, Western blot, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blotting, immunodetection method, surface plasmon resonance, optical spectroscopy, mass spectrometry, or HPLC.

[0275] Normalization of Expression Level In some embodiments, the expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, and / or MARCO are normalized expression levels. For example, the TAM signature score is the average of the normalized expression levels of one or more genes in a sample from an individual.

[0276] In some embodiments, the TAM signature score is the average of the normalized expression levels of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from an individual.

[0277] The detected expression levels of the genes can be normalized using any one of the standard normalization methods known in the art. One of ordinary skill in the art will understand that the normalization method used can be selected according to the gene expression methodology employed (e.g., one or more housekeeping genes can be used for normalization from the perspective of RT-qPCR methodology, while the entire genome or substantially the entire genome can be used as a normalization baseline from the perspective of RNA-seq methodology). For example, the detected expression levels of each assayed gene can be normalized for differences in the amount of the assayed gene(s), variations in the quality of the samples used, and / or variations between assay runs.

[0278] In some examples, normalization can be achieved by detecting the expression of one or more specific normalization genes, including reference gene(s) (e.g., housekeeping gene(s) such as β-actin). For example, in some examples, the 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., 1, 2, 3, 4, 5, 6, 7, 8, 9, or more reference genes, such as housekeeping genes (e.g., β-actin)). Alternatively, normalization can be performed based on the mean or median value of the signals of all assayed genes. For each gene, the measured amount of normalized mRNA can be compared to the amount found at the reference expression level. The measured presence and / or expression level / amount in a particular subject sample being analyzed is represented as a certain percentile within this range, which can be determined by methods well known in the art.

[0279] In other examples, the detected expression levels of each assayed gene are not normalized to determine the expression levels.

[0280] The expression level of each gene can be determined using any statistical approach known in the art. For example, the expression level can reflect a central expression level, a centrally normalized expression level, or an average expression level, or an average normalized expression level.

[0281] In some embodiments, the TAM signature score is a numerical value that reflects the aggregated Z-score expression levels for a combination of assayed genes (e.g., a combination of two or more of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO, e.g., a combination of all six of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO).

[0282] Any of the methods provided above may further include detecting additional genes in a sample from an individual, and may include, for example, detecting the expression levels of at least one and one or more additional genes of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO. In some embodiments, the method further includes detecting the expression levels of one or more of 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), progranulin (GRN), glioma pathogenesis-related protein 2 (GLIPR2), arrestin domain-containing protein 4 (ARRDC4), apolipoprotein E (APOE), folate receptor beta (FOLR2), and cathepsin D (CTSD) in a sample from an individual. In some examples, the method further includes detecting the expression levels of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 all of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD.

[0283] In some embodiments, determining the TAM signature score in a sample from an individual comprises further detecting the expression levels of all 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD.

[0284] For example, in some embodiments, the TAM signature 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 a sample from an individual. Measurement and normalization of the expression levels may be performed as described above. For example, in some embodiments, the TAM signature score is a numerical value reflecting the aggregated Z-score expression levels for combinations of 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.

[0285] In some embodiments, the method comprises further detecting the expression level of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD in a sample from an individual and determining a 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.

[0286] An individual is determined to have a TAM signature score that exceeds a reference TAM signature score, whereby, in some embodiments where the individual is identified as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, an expression level of one or more of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD has been detected in a sample from the individual.

[0287] In some embodiments where an individual is determined to have a TAM signature score that exceeds a reference TAM signature score, whereby the individual is identified as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, an expression level of each of ACP5, MCEMP1, CYP27A1, OLR1, GRN, GLIPR2, ARRDC4, APOE, FOLR2, and CTSD has been detected in a sample from the individual, from which the TAM signature score is determined, and 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 a sample from the individual.

[0288] Reference Expression Level and TAM Signature Score The terms “reference expression level” and “reference TAM signature score” refer to an expression level or TAM signature score to which another expression level or TAM signature score is compared, for example, to make a diagnosis, prediction, prognosis, and / or treatment determination.

[0289] In some embodiments, the reference expression level or reference TAM signature score is a pre-assigned reference expression level or reference TAM signature score.

[0290] In some embodiments, the reference expression level or reference TAM signature score is the expression level or TAM signature score in a reference population (e.g., a population of individuals having cancer, e.g., a population of individuals having lung cancer (e.g., NSCLC)).

[0291] In some embodiments, the expression level or TAM signature score of the reference population is the median of the expression level or TAM signature score of the reference population.

[0292] In other embodiments, the expression level or TAM signature score of the reference population is the average expression level or TAM signature score of the reference population.

[0293] In yet other embodiments, the expression level or TAM signature score is defined as the 25th percentile, 26th percentile, 27th percentile, 28th percentile, 29th percentile, 30th percentile, 31st percentile, 32nd percentile, 33rd percentile, 34th percentile, 35th percentile, 36th percentile, 37th percentile, 38th percentile, 39th percentile, 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, 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 of the expression level or TAM signature score in a reference population.

[0294] In some examples, the baseline expression level or baseline TAM signature score is a cut-off value that significantly separates first and second subsets of individuals treated with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., atezolizumab and tiragolumab) in the same reference population, based on the significant difference between the responsiveness of the individuals to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody above or below the cut-off value. In some embodiments, the responsiveness of an individual to treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody is significantly improved compared to the responsiveness of the individual to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody at or above the cut-off value.

[0295] PD-L1 status In some embodiments, the expression level of PD-L1 is being evaluated in a sample from a subject described herein. In some embodiments, the sample is determined to have a PD-L1 positive tumor cell fraction (e.g., by immunohistochemistry (IHC) assay, e.g., by positive staining with an anti-PD-L1 antibody (the anti-PD-L1 antibody is SP263, 22C3, SP142, or 28-8)).

[0296] In some embodiments, the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining using the anti-PD-L1 antibody SP263 (e.g., calculated using the Ventana SP263 IHC assay).

[0297] In some embodiments, the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining using the anti-PD-L1 antibody 22C3 (e.g., calculated using the pharmDx 22C3 IHC assay).

[0298] Exemplary methods for evaluating the expression level of PD-L1 are provided in Section III(E).

[0299] B. Bone marrow markers in serum samples during treatment Method for monitoring response to treatment In another aspect, the present invention is a method for monitoring the response of an individual having cancer (e.g., lung cancer, e.g., NSCLC) to a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV of this specification and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV of this specification), comprising detecting the expression level (e.g., gene expression level, e.g., protein expression level or nucleic acid expression level) of one or more of myeloid marker macrophage receptor with collagenous structure (MARCO), cathelicidin antimicrobial peptide (CAMP), CD5 antigen-like (CD5L), scavenger receptor cysteine-rich type 1 protein M130 (CD163), neutrophil gelatinase-associated lipocalin (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 E (APOE), lactotransferrin (TRFL), vascular cell adhesion protein 1 (VCAM1), PERM, beta2-microglobulin (B2MG), LYSC, LYAM1, LCAT, and LIRA3 in a sample from the individual during or after administration of the PD-1 axis-binding antagonist antibody and the anti-TIGIT antagonist antibody, and 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., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,Also provided is a method for predicting an individual who is likely to respond to treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody based on an increase in the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or all 20) of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3.

[0300] In some embodiments, 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 level, thereby predicting that the individual is likely to respond to treatment with a PD-1 axis-binding antagonist and an 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.

[0301] In some embodiments, an increase in the expression level (e.g., gene expression level, e.g., protein expression level or nucleic acid expression level) of MARCO, CAMP, CD5L, CD163, NGAL, CSF1R, CD44, APOC2, APOC3, APOC4, APOA2, APOE, TRFL, VCAM1, PERM, B2MG, LYSC, LYAM1, LCAT, and LIRA3 is defined as an increase of at least 1.2-fold (e.g., 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) relative to the reference level. For example, in some embodiments, the increased expression level is an expression level that increases 1.2-fold to 5-fold (e.g., 1.2-fold to 4-fold, 1.2-fold to 3-fold, 1.2-fold to 2-fold, 1.2-fold to 1.9-fold, 1.2-fold to 1.7-fold, or 1.2-fold to 1.5-fold).

[0302] In some embodiments, the response to treatment is an increase in progression-free survival (PFS) or overall survival (OS).

[0303] In some embodiments of any of the methods provided herein, the cancer is lung cancer, e.g., NSCLC. In some embodiments, the individual is human.

[0304] Expression level 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 detected by the methods provided herein can be a gene expression level, such as a nucleic acid expression level or a protein expression level (e.g., a protein level determined by mass spectrometry). Exemplary methods for detecting and normalizing nucleic acid expression levels are provided in Section IIIA above.

[0305] In some embodiments, the expression level is a protein expression level, such as a protein expression level determined by mass spectrometry, western blot, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blotting, immunodetection, surface plasmon resonance, optical spectroscopy, mass spectrometry or HPLC. In some embodiments, the protein expression level is measured in a serum sample.

[0306] In other embodiments, the expression level is a nucleic acid expression level, such as an mRNA expression level. In one embodiment, 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 in cells derived from a blood sample of an individual, such as peripheral blood mononuclear cells (PBMCs) derived from a blood sample of an individual.

[0307] Sample 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 can be determined from any suitable sample. Exemplary sample types include, but are not limited to, tissue samples, tumor samples, whole blood samples, plasma samples, serum samples, and combinations thereof. The sample may be fresh, stored, or frozen.

[0308] In some embodiments, the sample is a serum sample.

[0309] A sample (e.g., a serum sample) can be collected from an individual and 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 can be detected therein at any suitable time point after the first administration of a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody to the individual. For example, the sample can be collected 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, 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 after the start of a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (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 after the start of the treatment). In other examples, the sample is collected about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, or more than 8 weeks after the start of a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, or more than 8 weeks after the start of the treatment).

[0310] In some embodiments, 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 3 weeks after the initiation of treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a sample (e.g., a serum sample) is collected from an individual 3 weeks after the initiation of treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and 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 therein).

[0311] In some embodiments, 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 6 weeks after the initiation of treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a sample (e.g., a serum sample) is collected from an individual 6 weeks after the initiation of treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and 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 therein).

[0312] Reference expression level The term "reference expression level" refers to an expression level (e.g., a protein expression level) to which another expression level is compared, for example, to make a diagnosis, prediction, prognosis, and / or treatment determination.

[0313] In some embodiments, the baseline expression level is the level of expression in a sample from an individual (e.g., a sample collected from the individual immediately prior to the first administration of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, or at least 1 day, at least 1 week, or at least 1 month prior to the first administration of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody) at a time point prior to the initiation of treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

[0314] In some embodiments, the baseline expression level is a pre-assigned baseline expression level.

[0315] In some embodiments, the baseline expression level is the level of expression in a reference population (e.g., a population of individuals with cancer, e.g., a population of individuals with lung cancer (e.g., NSCLC)).

[0316] In some embodiments, the level of expression in the reference population is the median level of expression in the reference population.

[0317] In other embodiments, the level of expression in the reference population is the average level of expression in the reference population.

[0318] In yet another aspect, the expression level is defined as the 25th percentile, 26th percentile, 27th percentile, 28th percentile, 29th percentile, 30th percentile, 31st percentile, 32nd percentile, 33rd percentile, 34th percentile, 35th percentile, 36th percentile, 37th percentile, 38th percentile, 39th percentile, 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, 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 of the expression level in a reference population.

[0319] In some examples, the baseline expression level is a cut-off value that significantly separates first and second subsets of individuals treated with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., atezolizumab and tiragolumab) in the same reference population based on a significant difference between the responsiveness of the individuals to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody above or below the cut-off value. In some embodiments, the responsiveness of an individual to treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody is significantly improved compared to the responsiveness of the individual to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody at or above the cut-off value.

[0320] PD-L1 status In some embodiments, the expression level of PD-L1 is being evaluated in a sample from a subject as described herein. In some embodiments, the sample is determined to have a PD-L1 positive tumor cell fraction (e.g., by immunohistochemistry (IHC) assay, e.g., by positive staining with an anti-PD-L1 antibody (the anti-PD-L1 antibody is SP263, 22C3, SP142, or 28-8)).

[0321] In some embodiments, the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining using the anti-PD-L1 antibody SP263 (e.g., calculated using the Ventana SP263 IHC assay).

[0322] In some embodiments, the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining using the anti-PD-L1 antibody 22C3 (e.g., calculated using the pharmDx 22C3 IHC assay).

[0323] Exemplary methods for evaluating the expression level of PD-L1 are provided in Section III(E).

[0324] C. Regulatory T cell (Treg) genes and signatures A method of identifying an individual who may benefit from treatment (i) Treg genes In one aspect, the present invention provides a method of identifying an individual having cancer (e.g., lung cancer, e.g., non-small cell lung cancer (NSCLC)) who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein), the method comprising detecting the expression level of one or more (e.g., one, two, three, four, five, six, or all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2) of the Treg-related genes forkhead box protein P3 (FOXP3), cytotoxic T lymphocyte protein 4 (CTLA4), interleukin 10 (IL10), tumor necrosis factor receptor superfamily member 18 (TNFRSF18), C-C chemokine receptor type 8 (CCR8), zinc finger protein Eos (IKZF4), and zinc finger protein Helios (IKZF2) in a sample from the individual, and identifying the individual as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody if FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 each exceed their respective reference expression levels.

[0325] (ii) Treg signature score In another aspect, the present invention provides a method of identifying an individual having cancer (e.g., lung cancer, e.g., NSCLC) who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein), the method comprising detecting the expression levels of at least two (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 determining a regulatory T cell (Treg) signature score therefrom, wherein an individual having a Treg signature score above a reference Treg signature score is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0326] In another aspect, the present invention provides a method of identifying an individual having cancer (e.g., lung cancer, e.g., NSCLC) who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein), 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 an individual having a Treg signature score above a reference Treg signature score is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0327] In some embodiments, the individual has a Treg signature score in a sample that exceeds a reference Treg 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. Exemplary methods for determining a Treg signature score and an exemplary reference Treg signature score are provided below. Exemplary PD-1 axis-binding antagonists, anti-TIGIT antagonist antibodies, and methods of treatment comprising these agents are provided in Section IV.

[0328] In some embodiments of any of the methods provided herein, the cancer is lung cancer, such as NSCLC. In some embodiments, the individual is human.

[0329] Method for selecting a treatment (i) Treg gene In another aspect, the invention is a method for selecting a treatment for an individual having cancer (e.g., lung cancer, such as NSCLC), the method comprising detecting in a sample from the individual the expression level of one or more (e.g., one, two, three, four, five, six, or all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2) of the Treg-related genes FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2, and an expression level of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 that exceeds each respective reference expression level 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., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0330] (ii) Treg signature score In another aspect, the present invention provides a method for selecting a treatment method for an individual having cancer (e.g., lung cancer, e.g., 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 signature score therefrom, wherein an individual having a Treg signature score above a reference Treg signature score is identified as an individual who may benefit from a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0331] In another aspect, the present invention provides a method for selecting a treatment method for an individual having cancer (e.g., lung cancer, e.g., 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 signature score therefrom, wherein an individual having a Treg signature score above a reference Treg signature score is identified as an individual who may benefit from a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0332] In some embodiments, the individual has a Treg signature score in a sample that exceeds a baseline Treg 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.

[0333] Treatment method (i) Treg gene In another aspect, the invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), the method comprising: (a) detecting in a sample from the individual the expression level of one or more (e.g., one, two, three, four, five, six, or all seven) of the Treg-related genes FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2, wherein the expression level of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 exceeds the respective baseline expression level, thereby identifying the individual as an individual who may benefit from treatment with 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 (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0334] In another aspect, the present invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein), wherein the individual is determined to have an expression level of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 that exceeds a respective reference expression level, thereby identifying the individual as an individual who may benefit from treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

[0335] (ii) Treg signature score In another aspect, the present invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), comprising: (a) detecting the expression levels of at least two (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 determining therefrom a Treg signature score, wherein the Treg signature score exceeds a reference Treg signature score, thereby identifying the individual as an individual who may benefit from treatment with 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., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0336] In another aspect, the present invention is a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), 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 exceeds a reference Treg signature score, thereby identifying the individual as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, 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; and (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein).

[0337] In another aspect, the present invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein), wherein the individual has been determined to have a Treg signature score that exceeds a reference Treg signature score, thereby identifying the individual as an individual who may benefit from treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, and wherein the Treg signature score is based on the expression levels of at least two (e.g., two, three, four, five, six, or all seven) of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 detected in a sample from the individual.

[0338] In another aspect, the present invention provides a method of treating an individual having cancer (e.g., lung cancer, e.g., NSCLC), the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., a PD-1 axis-binding antagonist (e.g., atezolizumab) disclosed in Section IV herein and an anti-TIGIT antagonist antibody (e.g., tiragolumab) disclosed in Section IV herein), wherein the individual has been determined to have a Treg signature score that exceeds a reference Treg signature score, thereby identifying the individual as an individual who may benefit from treatment with the PD-1 axis-binding antagonist and the 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.

[0339] Advantage In some embodiments, the benefits achieved by treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody include an increase in progression-free survival (PFS) (e.g., an increase in the duration of PFS experienced by an individual treated according to the method or an increase in the average PFS of a population of individuals treated according to the method), an increase in objective response rate (ORR) (e.g., an increase in the ORR of a population of individuals treated according to the method), and / or an increase in overall survival (OS) (e.g., an increase in the duration of OS experienced by an individual treated according to the method or an increase in the average OS of a population of individuals treated according to the method).

[0340] The increased PFS, ORR, and / or OS can be determined, for example, by comparison to untreated reference individuals and / or a reference population of individuals, a control treatment, e.g., one or more previously approved treatments or reference individuals and / or a reference population of individuals who received a commercial product for the treatment of cancer; and / or reference individuals and / or a reference population of individuals treated with a PD-1 axis-binding antagonist (e.g., atezolizumab) or an anti-TIGIT antagonist antibody (e.g., tiragolumab) as monotherapy. In some embodiments, the increased PFS, ORR, and / or OS is determined by comparison to a reference individual and / or a reference population of individuals having cancer treated with a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., atezolizumab and tiragolumab), wherein each individual in the reference individual and / or reference population has a Treg signature score below a reference Treg signature score and / or has an expression level of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 below their respective reference expression levels. The reference Treg signature score is as described herein and can be, for example, the median of the Treg signature scores of a reference population of individuals having cancer (e.g., lung cancer, e.g., NSCLC) or the median of the expression levels of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a reference population of individuals having cancer (e.g., lung cancer, e.g., NSCLC).

[0341] An exemplary method for determining whether a given clinical outcome has been improved in accordance with the present invention is provided in Section III(A).

[0342] Sample The expression level and / or Treg signature score of one or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 can be determined from any suitable sample. Exemplary sample types include, but are not limited to, tissue samples, tumor samples, whole blood samples, plasma samples, serum samples, and combinations thereof. The sample may be fresh, stored, or frozen.

[0343] In some embodiments, the sample is a tissue sample, such as a tumor tissue sample. In some embodiments, the tumor tissue sample is a biopsy material. In some embodiments where the cancer is lung cancer (e.g., NSCLC), the sample is a biopsy material of lung cancer.

[0344] In some embodiments, the sample is obtained from an individual prior to treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, e.g., immediately prior to the first administration of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody, or at least 1 day, at least 1 week, or at least 1 month prior to the first administration of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

[0345] Treg signature score In some embodiments, determining a Treg signature score in a sample from an individual comprises calculating an average of the expression levels of at least two of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in the sample from the individual. Thus, in some embodiments, the Treg signature score is an average of the expression levels (e.g., the average of the normalized expression levels) of at least two of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from an individual.

[0346] In some embodiments, determining a Treg signature score in a sample from an individual comprises calculating an average of the expression levels of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in the sample from the individual. Thus, in some embodiments, the TAM signature score is an average of the expression levels (e.g., the average of the normalized expression levels) of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2 in a sample from an individual.

[0347] Expression level The expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and / or IKZF2 detected by the methods provided herein can be, for example, nucleic acid expression levels or protein expression levels (e.g., protein levels determined by mass spectrometry).

[0348] In some embodiments, the expression level is a nucleic acid expression level, e.g., an mRNA expression level. The nucleic acid expression level can be detected using any suitable method known in the art, e.g., 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 a combination thereof. Further exemplary methods for measuring nucleic acid expression levels are provided in Section III(A).

[0349] In some embodiments, the expression level is a protein expression level, e.g., a protein expression level determined by mass spectrometry, Western blot, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot blotting, immunodetection methods, surface plasmon resonance, optical spectroscopy, mass spectrometry or HPLC.

[0350] Normalization of expression levels In some embodiments, the expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and / or IKZF2 are normalized expression levels, e.g., the Treg signature score is the average of the normalized expression levels of one or more genes in a sample from an individual.

[0351] In some embodiments, the Treg signature score is the average of the normalized expression levels of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 in a sample from an individual.

[0352] Exemplary methods for normalizing the detected expression levels of genes are provided in Section III(A).

[0353] In some embodiments, the Treg signature score is a numerical value that reflects the aggregated Z-score expression levels for a combination of genes assayed (e.g., a combination of two or more of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2, e.g., a combination of all seven of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4, and IKZF2).

[0354] Reference Expression Level and Treg Signature Score The terms "reference expression level" and "reference Treg signature score" refer to an expression level or Treg signature score to which another expression level or Treg signature score is compared, for example, to make a diagnosis, prediction, prognosis, and / or treatment determination.

[0355] In some embodiments, the reference expression level or reference Treg signature score is a pre-assigned reference expression level or reference Treg signature score.

[0356] In some embodiments, the reference expression level or reference Treg signature score is the expression level or Treg signature score in a reference population (e.g., a population of individuals having cancer, e.g., a population of individuals having lung cancer (e.g., NSCLC)).

[0357] In some embodiments, the expression level or Treg signature score of the reference population is the median of the expression level or Treg signature score of the reference population.

[0358] In other embodiments, the expression level or Treg signature score of the reference population is the average expression level or Treg signature score of the reference population.

[0359] In yet other embodiments, the expression level or Treg signature score is defined as the 25th percentile, 26th percentile, 27th percentile, 28th percentile, 29th percentile, 30th percentile, 31st percentile, 32nd percentile, 33rd percentile, 34th percentile, 35th percentile, 36th percentile, 37th percentile, 38th percentile, 39th percentile, 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, 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 of the expression level or Treg signature score in a reference population.

[0360] In some examples, the baseline expression level or baseline Treg signature score is a cutoff value that significantly separates first and second subsets of individuals treated with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody (e.g., atezolizumab and tiragolumab) in the same reference population based on a significant difference between the responsiveness of individuals to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody above or below the cutoff value. In some embodiments, the responsiveness of an individual to treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody is significantly improved compared to the responsiveness of an individual to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody above the cutoff value.

[0361] PD-L1 status In some embodiments, the expression level of PD-L1 is being evaluated in a sample from a subject described herein. In some embodiments, the sample is determined to have a PD-L1 positive tumor cell fraction (e.g., by immunohistochemistry (IHC) assay, e.g., by positive staining with an anti-PD-L1 antibody (the anti-PD-L1 antibody is SP263, 22C3, SP142, or 28-8)).

[0362] In some embodiments, the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining with the anti-PD-L1 antibody SP263 (e.g., calculated using the Ventana SP263 IHC assay).

[0363] In some embodiments, the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining with the anti-PD-L1 antibody 22C3 (e.g., calculated using the pharmDx 22C3 IHC assay).

[0364] Exemplary methods for evaluating the expression level of PD-L1 are provided in Section III(E).

[0365] Evaluation of TIGIT Expression In some embodiments, the expression of TIGIT is evaluated in the individuals described herein. The methods provided herein can include determining the expression level of TIGIT in a biological sample (e.g., a tumor sample) obtained from an individual. In other examples, the expression level of TIGIT in a biological sample (e.g., a tumor sample) obtained from an individual has been determined before or after the start of treatment. TIGIT expression can be determined using any suitable method. Any suitable tumor sample can be used, such as a formalin-fixed paraffin-embedded (FFPE) tumor sample, a stored tumor sample, a fresh tumor sample, or a frozen tumor sample.

[0366] For example, the expression of TIGIT can be determined as the proportion of tumor-infiltrating immune cells in a tumor sample that express a detectable expression level of TIGIT, with respect to the proportion of the tumor sample covered by tumor-infiltrating immune cells, and / or as the proportion of tumor cells in a tumor sample that express a detectable expression level of TIGIT. In any of the foregoing examples, the proportion of the tumor sample composed of tumor-infiltrating immune cells can be, for example, with respect to the proportion of the tumor area covered by tumor-infiltrating immune cells in a section of the tumor sample obtained from an individual, when evaluated 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 / 126688 A3; 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 embodiments, the TIGIT antagonist antibody (e.g., SP410) is detected using the VENTANA OptiView DAB IHC Detection Kit on an automated VENTANA BenchMark ULTRA platform.

[0367] E. Evaluation of PD-L1 Expression In some embodiments, the expression of PD-L1 is evaluated in the individuals described herein. The methods provided herein can include determining the expression level of PD-L1 in a biological sample (e.g., a tumor sample) obtained from an individual. In other examples, the expression level of PD-L1 in a biological sample (e.g., a tumor sample) obtained from an individual has been determined before or after the start of treatment. PD-L1 expression can be determined using any suitable approach. For example, PD-L1 expression can be determined as described in U.S. Patent Application Publication No. 20180030138 A1 and U.S. Patent Application Publication No. 20180037655 A1. Any suitable tumor sample can be used, such as a formalin-fixed paraffin-embedded (FFPE) tumor sample, a stored tumor sample, a fresh tumor sample, or a frozen tumor sample.

[0368] For example, the expression of PD-L1 can be determined as a percentage of tumor-infiltrating immune cells in a tumor sample that express PD-L1 at a detectable expression level, with respect to the percentage of the tumor sample composed of tumor-infiltrating immune cells that express PD-L1 at a detectable expression level, and / or as a percentage of tumor cells in a tumor sample that express PD-L1 at a detectable expression level. In any of the foregoing examples, the percentage of the tumor sample composed of tumor-infiltrating immune cells can be understood to be a percentage related to the percentage of the tumor area covered by tumor-infiltrating immune cells in a section of the tumor sample obtained from an individual, which is evaluated by IHC using, for example, an anti-PD-L1 antibody (e.g., the SP142 antibody). It should be understood that any suitable anti-PD-L1 antibody can be used, including, for example, SP142 (Ventana), SP263 (Ventana), 22C3 (Dako), 28-8 (Dako), E1L3N (Cell Signaling 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 examples, the anti-PD-L1 antibody is 28-8.

[0369] In some examples, the tumor sample obtained from an individual has PD-L1 at a detectable expression level in less than 1% of the tumor cells in the tumor sample, in 1% or more of the tumor cells in the tumor sample, in 1% to less than 5% of the tumor cells in the tumor sample, in 5% or more of the tumor cells in the tumor sample, in 5% to less than 50% of the tumor cells in the tumor sample, or in 50% or more of the tumor cells in the tumor sample.

[0370] In some examples, the tumor sample obtained from an individual has a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise less than 1% of the tumor sample, more than 1% of the tumor sample, less than 1% to 5% of the tumor sample, more than 5% of the tumor sample, less than 5% to 10% of the tumor sample, or more than 10% of the tumor sample.

[0371] In some embodiments, the tumor sample obtained from an individual has a detectable PD-L1 expression level in tumor-infiltrating immune cells that comprise 5% to 19% (e.g., TIC 5% to 19%) of the tumor sample, e.g., a PD-L1 expression level that is low for PD-L1. In some embodiments, the tumor sample obtained from an individual has a detectable PD-L1 expression level in tumor-infiltrating immune cells that comprise ≧20% (e.g., TIC ≧20%) of the tumor sample, e.g., a PD-L1 expression level that is high for PD-L1. In some embodiments, tumor samples determined to have 5% or more TIC are comparable to one or more CPS.

[0372] In some examples, the tumor sample can be scored for PD-L1 positivity in tumor-infiltrating immune cells and / or tumor cells according to the criteria for diagnostic assessment shown in Table 1 and / or Table 2, respectively. [Table 1] [Table 2]

[0373] In some examples, in any of the methods, uses or compositions for use described herein, the individual has a PD-L1 selected tumor (e.g., the percentage of tumor area occupied by PD-L1 expressing tumor infiltrating immune cells (IC) is 5% or more in a tumor sample as determined by IHC using the SP 142 antibody). In some examples, a PD-L1 selected tumor is a tumor determined by an immunohistochemistry (IHC) assay to have a percentage of tumor area occupied by PD-L1 expressing immune cells (IC) of 5% or more. In some examples, the IHC assay uses an anti-PD-L1 antibody SP142, SP263, 22C3 or 28-8. In some examples, the IHC assay uses the anti-PD-L1 antibody SP142. In some examples, the IHC assay uses the anti-PD-L1 antibody SP263. In some examples, the IHC assay uses the anti-PD-L1 antibody 22C3. In some examples, the IHC assay uses the anti-PD-L1 antibody 22C3. In some examples, the IHC assay uses the anti-PD-L1 antibody 28-8.

[0374] In some examples, the IC score is determined to be 5% or more (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some examples, the IC score is determined to be 2 or 3 (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some examples, the IC score is determined to be 1% or more (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some examples, the IC score is determined to be 10% or more (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some examples, the IC score is determined to be 1% or more and less than 50% (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay). In some examples, the IC score is determined to be 1% or more and less than 30% (e.g., as determined using the Ventana (SP142) PD-L1 IHC assay).

[0375] In some examples, in any of the methods, uses, or compositions for use described herein, a tumor sample obtained from an individual has a detectable PD-L1 protein expression level. In some examples, the detectable PD-L1 protein expression level is determined by an IHC assay. In some examples, the IHC assay uses the anti-PD-L1 antibody SP142. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in tumor-infiltrating immune cells that constitute 5% or more of the tumor sample. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in tumor-infiltrating immune cells that constitute 1% or more of the tumor sample. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in tumor-infiltrating immune cells that constitute more than 1% and less than 5% of the tumor sample. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in tumor-infiltrating immune cells that constitute 5% or more and less than 10% of the tumor sample. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in tumor-infiltrating immune cells that constitute 10% or more of the tumor sample. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in 1% or more of the tumor cells in the tumor sample. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in 1% or more and less than 5% of the tumor cells in the tumor sample. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in 5% or more and less than 50% of the tumor cells in the tumor sample. In some examples, the tumor sample is determined to have a detectable PD-L1 expression level in 50% or more of the tumor cells in the tumor sample.

[0376] In some examples, in any of the methods, uses, or compositions for use described herein, an individual has a PD-L1 selected tumor (e.g., a PD-L1 “high” selected tumor (e.g., a tumor proportion score (TPS) of PD-L1 in a tumor sample of greater than 50% as determined by IHC using the SP 263 antibody)). In some examples, the PD-L1 selected tumor is a PD-L1 “high” selected tumor. In some examples, the PD-L1 selected tumor is a tumor determined to have a TPS of greater than 50% by an immunohistochemistry (IHC) assay. In some examples, the IHC assay uses an anti-PD-L1 antibody SP263, SP142, 22C3, or 28-8. In some examples, the IHC assay uses the anti-PD-L1 antibody SP263. In some examples, the IHC assay uses the anti-PD-L1 antibody SP142. In some examples, the IHC assay uses the anti-PD-L1 antibody 22C3. In some examples, the TPS is determined to be greater than or equal to 50% (e.g., when determined using the Ventana (SP263) PD-L1 IHC assay). In some examples, the TPS is determined to be less than 50% (e.g., when determined using the Ventana (SP263) PD-L1 IHC assay). In some examples, the TPS is determined to be greater than or equal to 1% (e.g., when determined using the Ventana (SP263) PD-L1 IHC assay). In some examples, the TPS is determined to be greater than or equal to 1% and less than 50% (e.g., when determined using the Ventana (SP263) PD-L1 IHC assay).

[0377] In some examples, in any of the methods, uses, or compositions for use described herein, a tumor sample obtained from an individual has a detectable PD-L1 protein expression level. In some examples, the detectable PD-L1 protein expression level is determined by an IHC assay. In some examples, the IHC assay uses the anti-PD-L1 antibody SP263. In some examples, the tumor sample has been determined to have a PD-L1 positive tumor cell fraction of 50% or more of the tumor sample. In some examples, the tumor sample has been determined to have a PD-L1 positive tumor cell fraction of less than 50% of the tumor sample. In some examples, the tumor sample has been determined to have a PD-L1 positive tumor cell fraction of 1% or more and less than 50% of the tumor sample.

[0378] In some examples, the IHC assay uses the anti-PD-L1 antibody 22C3. In some examples, the IHC assay is the pharmDx 22C3 IHC assay. In some examples, the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining using the anti-PD-L1 antibody 22C3. In some embodiments, the tumor sample has been determined to have a combined positive score (CPS) of 10 or more or a tumor proportion score (TPS) of 1% or more in the tumor sample when determined using the anti-PD-L1 antibody 22C3, for example, as part of the pharmDx 22C3 IHC assay. In some embodiments, the tumor sample has been determined to have a CPS of 10 or more or a TPS of 1% or more and less than 50% in the tumor sample as determined using the anti-PD-L1 antibody 22C3, for example, as part of the pharmDx 22C3 IHC assay. In some embodiments, the tumor sample has been determined to have a CPS of 20 or more or a TPS of 50% or more in the tumor sample as determined using the anti-PD-L1 antibody 22C3, for example, as part of the pharmDx 22C3 IHC assay. In some embodiments, a tumor sample determined to have one or more CPS is comparable to 5% or more of the TIC.

[0379] In some examples, the IHC assay uses the anti-PD-L1 antibody 28-8. In some examples, the IHC assay is the pharmDx 28-8 IHC assay. In some examples, the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining using the anti-PD-L1 antibody 28-8.

[0380] In some examples, in any of the methods, uses, or compositions for use described herein, a tumor sample obtained from an individual has a detectable PD-L1 nucleic acid expression level. In some examples, the detectable PD-L1 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. In some examples, the sample is selected from the group consisting of a tissue sample, a whole blood sample, a serum sample, and a plasma sample. In some examples, the tissue sample is a tumor sample. In some examples, the tumor sample comprises tumor infiltrating immune cells, tumor cells, stromal cells, and any combination thereof.

[0381] IV. Exemplary anti-TIGIT antagonist antibodies and PD-1 axis binding antagonists Exemplary anti-TIGIT antagonist antibodies and PD-1 axis binding antagonists useful for treating an individual (e.g., a human) having cancer according to the methods, uses, and compositions for use of the invention are described herein.

[0382] A. Exemplary anti-TIGIT antagonist antibodies The present invention provides an anti-TIGIT antagonist antibody useful for treating cancer in a subject (e.g., a human).

[0383] In some examples, the anti-TIGIT antagonist antibody is tiragolumab (CAS registration number: 1918185-84-8). Tiragolumab (Genentech) is also known as MTIG7192A.

[0384] In certain examples, the anti-TIGIT antagonist antibody comprises at least 1, 2, 3, 4, 5, or 6 HVRS selected from: (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 / or (f) HVR-L3 comprising the amino acid sequence of QQYYSTPFT (SEQ ID NO: 6), or one or more combinations of the foregoing HVRS, and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity) to any one of SEQ ID NOS: 1-6.

[0385] In some examples, any of the anti-TIGIT antagonist antibodies may include: (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). In some examples, the anti-TIGIT antagonist antibody has an amino acid sequence having at least 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity) with or the sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO: 17), or has an amino acid sequence having at least 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity) with or the sequence of QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSS (SEQ ID NO: 18), and / or has 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 or the sequence of DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAWYQQKPGQPPNLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPFTFGPGTKVEIK (SEQ ID NO: 19).In some examples, 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 SEQ ID NO: 17 or its sequence, 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 SEQ ID NO: 19 or its sequence. In some examples, 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 examples, 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 SEQ ID NO: 18 or its sequence, 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 SEQ ID NO: 19 or its sequence. In some examples, 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.

[0386] In some examples, the anti-TIGIT antagonist antibody comprises a heavy chain and a light chain sequence, wherein (a) the heavy chain has the amino acid sequence: EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGKTYYRFKWYSDYAVSVKGRITINPDTSKNQFSLQLNSVTPEDTAVFYCTRESTTYDLLAGPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 33); and (b) the light chain has the amino acid sequence: DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAWYQQKPGQPPNLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSTPFTFGPGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 34). In some embodiments, 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.

[0387] In some examples, the anti-TIGIT antagonist antibody further comprises at least one, two, three, or four of the following light chain variable region framework regions (FR): FR-L1 comprising the amino acid sequence of DIVMTQSPDSLAVSLGERATINC (SEQ ID NO: 7); FR-L2 comprising the amino acid sequence of WYQQKPGQPPNLLIY (SEQ ID NO: 8); FR-L3 comprising the amino acid sequence of GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 9); and / or FR-L4 comprising the amino acid sequence of FGPGTKVEIK (SEQ ID NO: 10), or one or more combinations of the above FRs, and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 7-10. In some examples, for example, the antibody further comprises FR-L1 comprising the amino acid sequence of DIVMTQSPDSLAVSLGERATINC (SEQ ID NO: 7), FR-L2 comprising the amino acid sequence of WYQQKPGQPPNLLIY (SEQ ID NO: 8), FR-L3 comprising the amino acid sequence of GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 9), and FR-L4 comprising the amino acid sequence of FGPGTKVEIK (SEQ ID NO: 10).

[0388] In some examples, the anti-TIGIT antagonist antibody further comprises one, two, three, or four of the following heavy chain variable region FRs: FR-H1 comprising the amino acid sequence of X1VQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 11) (where X1 is E or Q); FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 11-14. The anti-TIGIT antagonist antibody can further comprise, for example, at least one, two, three, or four of the following heavy chain variable region framework regions FR: FR-H1 comprising the amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 15); FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and / or FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14), or a combination of one or more of the above FRs and one or more variants thereof having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 12-15. In some examples, the anti-TIGIT antagonist antibody comprises FR-H1 comprising the amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 15); FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14).In another example, the anti-TIGIT antagonist antibody comprises at least one, two, three, or four of the following heavy chain variable region framework regions FR: FR-H1 comprising the amino acid sequence of QVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 16); FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and / or FR-H4 comprising 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 having at least about 90% sequence identity (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity) to any one of SEQ ID NOs: 12-14 and 16. In some examples, the anti-TIGIT antagonist antibody comprises FR-H1 comprising the amino acid sequence of QVQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 16), FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLG (SEQ ID NO: 12); FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLQLNSVTPEDTAVFYCTR (SEQ ID NO: 13); and FR-H4 comprising the amino acid sequence of WGQGTLVTVSS (SEQ ID NO: 14).

[0389] In another aspect, an anti-TIGIT antagonist antibody is provided, the antibody comprising a VH as in any of the above examples and a VL as in any of the above examples, wherein one or both of the variable domain sequences comprise post-translational modifications.

[0390] In some examples, any one of the above anti-TIGIT antagonist antibodies can bind to rabbit TIGIT in addition to human TIGIT. In some examples, any one of the above anti-TIGIT antagonist antibodies can bind to both human TIGIT and cynomolgus (cyno) TIGIT. In some examples, any one of the above anti-TIGIT antagonist antibodies can bind to human TIGIT, cynomolgus TIGIT, and rabbit TIGIT. In some examples, any one of the above anti-TIGIT antagonist antibodies can bind to human TIGIT, cynomolgus TIGIT, and rabbit TIGIT, but cannot bind to mouse TIGIT.

[0391] In some examples, the anti-TIGIT antagonist antibody binds to human TIGIT with a K D of about 10 nM or less and binds to cynomolgus TIGIT with a K D of about 10 nM or less (e.g., binds to human TIGIT with a K D of about 0.1 nM to about 1 nM and binds to cynomolgus TIGIT with a K D of about 0.5 nM to about 1 nM, and for example, binds to human TIGIT with a K D of about 0.1 nM or less and binds to cynomolgus TIGIT with a K D of about 0.5 nM or less).

[0392] In some examples, the anti-TIGIT antagonist antibody specifically binds to TIGIT and inhibits or blocks the TIGIT interaction with the poliovirus receptor (PVR) (e.g., the antagonist antibody inhibits the intracellular signaling mediated by TIGIT binding to PVR). In some examples, 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 examples, the anti-TIGIT antagonist antibody specifically binds to TIGIT and inhibits or blocks the TIGIT interaction with PVR without affecting the PVR-CD226 interaction. In some examples, the antagonist antibody inhibits or blocks the binding of cynomolgus TIGIT to cynomolgus PVR with an IC50 value of 50 nM or less (e.g., 1 nM to about 50 nM, e.g., 1 nM to about 5 nM). In some examples, the anti-TIGIT antagonist antibody inhibits and / or blocks the interaction between CD226 and TIGIT. In some examples, the anti-TIGIT antagonist antibody inhibits and / or blocks the ability of TIGIT to disrupt CD226 homodimerization.

[0393] In some examples, the methods or uses described herein may involve using or administering an isolated anti-TIGIT antagonist antibody that competes with any of the anti-TIGIT antagonist antibodies described above for binding to TIGIT. For example, the method may involve administering an isolated anti-TIGIT antagonist antibody that competes with an anti-TIGIT antagonist antibody having the following six HVRS for binding to TIGIT: (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). The methods described herein may also involve administering an isolated anti-TIGIT antagonist antibody that binds to the same epitope as the anti-TIGIT antagonist antibodies described above.

[0394] In some embodiments, the anti-TIGIT antagonist antibody is an antibody having intact Fc-mediated effector function (e.g., tiragolumab, vibostolimab, etigilimab, EOS084448 or TJ-T6) or enhanced effector function (e.g., SGN-TGT).

[0395] In some embodiments, the anti-TIGIT antagonist antibody comprises an Fc domain that can interact (e.g., activate) with an Fc gamma receptor (FcγR). In some embodiments, the anti-TIGIT antagonist antibody comprises an Fc domain that can interact (e.g., activate) with an FcγR of one or more myeloid cell types (e.g., one or more of cDC1, macrophages, neutrophils, and circulating monocytes).

[0396] In some embodiments, the anti-TIGIT antagonist antibody is capable of one or more Fc-dependent activations of one or more myeloid cell types, such as intratumoral type 1 conventional dendritic cells (cDC1), macrophages, neutrophils, and circulating monocytes.

[0397] In some embodiments, the anti-TIGIT antagonist antibody can interact with FcγR of one or more myeloid cell types (e.g., one or more of cDC1, macrophages, neutrophils, and circulating monocytes), and can induce CD8+ T cell mobilization in the blood and / or expansion of proliferating CD8+ T cells in the tumor bed.

[0398] In some embodiments, the anti-TIGIT antagonist antibody can interact with (e.g., upregulate) the MYC targeting pathway. In some embodiments, the present invention includes detecting the level (e.g., gene expression level, e.g., protein level or nucleic acid level) of one or more members of the MYC targeting pathway (e.g., MYC) in a sample from an individual, e.g., detecting the level of one or more members of the MYC targeting pathway in a sample from an individual at a time point during or after administration of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and comparing the detected level to a baseline expression level, e.g., a baseline expression level from a sample from the individual at a time point before initiation of treatment including a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

[0399] In some embodiments, the anti-TIGIT antagonist antibody is an IgG1 class antibody, such as tirzolgomab, vibostolimab, etigilimab, BGB-A1217, SGN-TGT, EOS084448 (EOS-448), TJ-T6, or AB308. Tirzolgomab is an anti-TIGIT antibody monoclonal antibody (mAb) having IgG1 / kappa Fc.

[0400] In other embodiments, the anti-TIGIT antagonist antibody is an IgG4 class antibody.

[0401] An anti-TIGIT antagonist antibody (e.g., tirzepatide) useful in the present invention, including a composition containing such an antibody, can be used in combination with a PD-1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antagonist antibody, e.g., atezolizumab), a PD-1 binding antagonist (e.g., an anti-PD-1 antagonist antibody, e.g., pembrolizumab) and a PD-L2 binding antagonist (e.g., an anti-PD-L2 antagonist antibody)).

[0402] In some embodiments, the anti-TIGIT antagonist antibody functions to inhibit TIGIT signaling. In some embodiments, the anti-TIGIT antagonist antibody inhibits 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, the anti-TIGIT antagonist antibody can inhibit the binding between TIGIT and CD155. In some embodiments, the anti-TIGIT antagonist antibody can inhibit the binding between TIGIT and CD112. In some embodiments, the anti-TIGIT antagonist antibody inhibits the binding between TIGIT and CD113. In some embodiments, the anti-TIGIT antagonist antibody inhibits TIGIT-mediated cell signaling in immune cells. In some embodiments, the anti-TIGIT antagonist antibody inhibits TIGIT by depleting regulatory T cells (e.g., FcγR).

[0403] 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.

[0404] In some embodiments, the anti-TIGIT antibody is selected from the group consisting of tirzolgomab (MTIG7192A, RG6058, or RO7092284), vibostolimab (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 tirzolgomab (MTIG7192A, RG6058, or RO7092284), vibostolimab (MK-7684), EOS-448, and SEA-TGT (SGN-TGT). The anti-TIGIT antibody can be tirzolgomab (MTIG7192A, RG6058, or RO7092284).

[0405] Non-limiting examples of anti-TIGIT antibodies useful in the methods disclosed herein and methods for their preparation are described in International Publication No. WO 2018 / 183889 A1, International Publication No. WO 2019 / 129261 A1, International Publication No. WO 2016 / 106302 A9, International Publication No. WO 2018 / 033798 A1, International Publication No. WO 2020 / 020281 A1, International Publication No. WO 2019 / 023504 A1, International Publication No. WO 2017 / 152088 A1, International Publication No. WO 2016 / 028656 A1, International Publication No. WO 2017 / 030823 A2, International Publication No. WO 2018 / 204405 A1, International Publication No. WO 2019 / 152574 A1 and International Publication No. WO 2020 / 041541 A2; U.S. Patent No. 10,189,902, U.S. Patent No. 10,213,505, U.S. Patent No. 10,124,061, U.S. Patent No. 10,537,633 and U.S. Patent No. 10,618,958;and those described in US Patent Application Publication Nos. 2020 / 0095324, 2019 / 0112375, 2018 / 0371083, and 2020 / 0062859, each of which is incorporated herein by reference. Further non-limiting examples of anti-TIGIT antibodies useful in the methods disclosed herein and methods for making them are International Publication Nos. WO 2018 / 204363 A1, WO 2018 / 047139 A1, WO 2019 / 175799 A2, WO 2018 / 022946 A1, WO 2015 / 143343 A2, WO 2018 / 218056 A1, WO 2019 / 232484 A1, WO 2019 / 079777 A1, WO 2018 / 128939 A1, WO 2017 / 196867 A1, WO 2019 / 154415 A1, WO 2019 / 062832 A1, WO 2018 / 234793 A3, WO 2018 / 102536 A1, WO 2019 / 137548 A1, WO 2019 / 129221 A1, WO 2018 / 102746 A1, WO 2018 / 160704 A9, WO 2020 / 041541 A2, WO 2019 / 094637 A9, WO 2017 / 037707 A1, WO 2019 / 168382 A1, WO 2006 / 124667 A3, WO 2017 / 021526 A1, WO 2017 / 184619 A2, WO 2017 / 048824 A1, WO 2019 / 032619 A9, WO 2018 / 157162 A1, WO 2020 / 176718 A1, WO 2020 / 047329 A1, WO 2020 / 047329 A1, WO 2018 / 220446 A9; US Patent Nos. 9,617,338, 9,567,399, 10,604,576, and US 9,994,637;and U.S. Patent Application Publication Nos. 2018 / 0355040, 2019 / 0175654, 2019 / 0040154, 2019 / 0382477, 2019 / 0010246, 2020 / 0164071, 2020 / 0131267, 2019 / 0338032, 2019 / 0330351, 2019 / 0202917, 2019 / 0284269, 2018 / 0155422, 2020 / 0040082, 2019 / 0263909, 2018 / 0185480, 2019 / 0375843, 2017 / 0037133, 2019 / 0077869, 2019 / 0367579, 2020 / 0222503, 2020 / 0283496, Chinese Patent Application Publication Nos. 109734806 A and 110818795 A, each of which is incorporated herein by reference.;

[0406] Anti-TIGIT antibodies useful in the methods disclosed herein include BGB-A1217, M6223, IBI939, EOS-448, vibostolimab (MK-7684) and SEA-TGT (SGN-TGT). Further anti-TIGIT antibodies useful in the methods disclosed herein include AGEN1307; AGEN1777; 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, MAB 7, MAB8, MAB9, MAB 10, MAB 11, MAB 12, MAB13, MAB 14, MAB 15, MAB 16, MAB 17, MAB 18, MAB19, MAB20, MAB21 (Astellas 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, (such antibodies having a modified heavy chain constant region (Bristol-Myers Squibb) included); 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 antibodies (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 Immunological Drugs 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-27278, ADI-30193, ADI-30296, ADI-27291, ADI-30283, ADI-30286, ADI-30288, ADI27297, ADI-30272, ADI-30278, ADI-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 (JN Biosciences 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); in-house anti-TIGIT (anti-PD-L1) native variable domain and Kymab TIGIT antigen-binding site (ABS) domain (bispecific 1) with bispecific antibody 1D05 / 1D05, In-house anti-TIGIT / 1D05 (bispecific 2) with Kymab TIGIT native variable domain and 1D05 ABS domain, Tool anti-TIGIT / Tool anti-PD-L1 (bispecific 3) with Toon anti-TIGIT native variable domain and Tool anti-PD-L1 ABS domain, Tool anti-PD-L1 / Tool anti-TIGIT (bispecific 4) with Tool anti-PD-L1 native variable domain and Tool anti-TIGIT ABS domain (Kymab 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); etigilimab (OMP-313M32) (Mereo BioPharma);Antibody clones 64G1E9B4, 100C4E7D11, 83G5H11C12, 92E9D4B4, 104G12E12G2, 121C2F10B5, 128E3F10F3F2, 70A11A8E6, 11D8E124A, 16F10H12C11, 8F2D8E7, 48B5G4E12, 139E2C2D2, 128E3G7F5, AS19584, AS19852, AS19858, AS19886, AS19887, AS19888, AS20160, AS19584VH26, AS19584VH29, AS19584VH30, AS19584VH31, AS19886VH5, 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, Ab29, Ab135, Ab171, Ab194, Ab184, Ab164, Ab183, Ab158, Ab55, Ab136, Ab39, Ab159, Ab151, Ab139, Ab107, Ab36, Ab193, Ab115, Ab106, Ab13f8, Ab127, Ab165, Ab155, Ab19, Ab6, Ab187, Ab179, Ab65, Ab114, 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, Ab150, 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, Ab172, 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, 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, 13 IgG2a defucosylated, 13 hIgG1 wild type, and 13 LALA-PG (Seattle Genetics); JS006 (Shanghai Junshi Biosciences Ltd.); anti-TIGIT Fc antibody and bispecific antibody PD1×TIGIT (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), 318.2...

Claims

**Claim 1** A method of identifying an individual having cancer who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, said method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from said individual and determining therefrom a tumor-associated macrophage (TAM) signature score, wherein a TAM signature score above a reference TAM signature score identifies said individual as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody. **Claim 2** A method for selecting a treatment method for an individual having cancer, said method comprising detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO in a sample from said individual and determining therefrom a TAM signature score, wherein a TAM signature score above a reference TAM signature score identifies said individual as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody. **Claim 3** The method according to claim 1 or 2, wherein said individual has a TAM signature score in said sample above a reference TAM signature score, and said method further comprises administering to said individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody. **Claim 4** A method of treating an individual having cancer, said method comprising (a) detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from said individual and determining therefrom a TAM signature score, wherein said TAM signature score is above a reference TAM signature score, whereby said individual is identified as an individual who may benefit from treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, detecting the expression level of each of C1QC, MSR1, MRC1, VSIG4, SPP1 and MARCO in a sample from said individual and determining therefrom a TAM signature score; (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody; A method comprising. **Claim 5** A method of treating an individual having cancer, the method comprising administering to the individual a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, wherein the individual is determined to have a TAM signature score that exceeds a reference TAM signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and the TAM signature score is based on the expression levels of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO detected in a sample from the individual. **Claim 6** The method according to any one of claims 1-5, wherein the sample is obtained from the individual prior to treatment with the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody. **Claim 7** The method according to any one of claims 1-6, wherein the benefit is an increase in progression-free survival (PFS), objective response rate (ORR), or overall survival (OS). **Claim 8** The method according to any one of claims 1-7, wherein the reference TAM signature score is a pre-assigned TAM signature score. **Claim 9** The method according to any one of claims 1-8, wherein the reference TAM signature score is the TAM signature score in a reference population. **Claim 10** The method according to claim 9, wherein the TAM signature score in the reference population is the median of the TAM signature scores of the reference population. **Claim 11** The method according to claim 9 or 10, wherein the reference population is a population of individuals having the cancer. **Claim 12** The method according to any one of claims 1-11, 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. **Claim 13** The method according to 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. **Claim 14** The method according to any one of claims 1 to 4 and 6 to 13, further comprising 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 according to 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 according to claim 14 or 15, further comprising 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 according to 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 according to 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 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, from which the TAM signature score is determined, and 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. The method according to claim 17 or 18.

20. A method for monitoring the response of an individual having cancer to a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, the method comprising detecting 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 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, and 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 each respective reference expression level predicts an individual likely to respond to the treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

21. The method according to 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 3 weeks after the start of the treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

22. The method according to 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 6 weeks after the start of the treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody.

23. The method according to 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 level, thereby predicting that the individual is likely to 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 according to any one of claims 20 to 23, wherein the response to the treatment is an increase in PFS or OS.

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

26. A method of identifying an individual having cancer who is likely to benefit from treatment with 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 an individual having a Treg signature score above a reference Treg signature score is identified as an individual who is likely to benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody.

27. A method for selecting a treatment method 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 an individual having a Treg signature score above a reference Treg signature score is identified 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 according to claim 26 or 27, wherein the individual has a Treg signature score in the sample above a reference Treg 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.

29. A method of treating an individual having 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 above a reference Treg signature score, whereby the individual is identified as an individual who may benefit from a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, 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; and (b) administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody. A method comprising.

30. A method of treating an individual having 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 signature score that exceeds a reference Treg signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and the Treg signature score is based on the expression levels of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in a sample from the individual.

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

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

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

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

36. The method according to claim 34 or 35, wherein the reference population is a population of individuals having the cancer.

37. The method according to 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 according to claim 37, 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.

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 according to claim 39, wherein the expression level is a nucleic acid expression level.

41. The method according to 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 according to claim 40 or 41, wherein the nucleic acid expression level is an mRNA expression level.

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

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

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

46. The method according to 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 according to claim 46, wherein the sample is a tissue sample.

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

49. The method according to claim 48, wherein the tumor tissue sample is a biopsy material.

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

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

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

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

54. The method according to claim 53, wherein when the PD-L1 positive tumor cell fraction is determined by positive staining using the anti-PD-L1 antibody SP263, it is 50% or more.

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

56. The method according to claim 53, wherein the PD-L1 positive tumor cell fraction is 50% or more when determined by positive staining using the anti-PD-L1 antibody 22C3.

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

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

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

60. The anti-TIGIT antagonist antibody has 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 ESTTYDLLAGPFDFY (SEQ ID NO: 3); (d) HVR-L1 comprising the amino acid sequence of KSSQTVLYSNNKKYLAA (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 QQYYSFTPFT (SEQ ID NO: 6) The method according to any one of claims 1 to 59.

61. The anti-TIGIT antagonist antibody further has the following light chain variable region FRs: (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 GVPDRFSSGSGSGTDFTLTISSLQAEDVAVYYC (SEQ ID NO: 9); and (d) FR-L4 comprising the amino acid sequence of FGPGTKVEIK (SEQ ID NO: 10) The method according to claim 60.

62. The anti-TIGIT antagonist antibody further has the following heavy chain variable region FRs: (a) X １ FR-H1 containing the amino acid sequence of VQLQQSGPGLVKPSQTLSLTCAISGDSVS (SEQ ID NO: 11), wherein X １ is Q or E); (b) FR-H2 comprising the amino acid sequence of WIRQSPSRGLEWLNG (SEQ ID NO: 12); (c) FR-H3 comprising the amino acid sequence of RITINPDTSKNQFSLLNSVTPEDTAVFCTR (SEQ ID NO: 13); and (d) FR-H4 comprising the amino acid sequence of WGQGTVTVSS (SEQ ID NO: 14) The method according to claim 60 or 61.

63. X １ The method according to claim 62, wherein X is Q.

64. X １ The method according to claim 62, wherein X is E.

65. The anti-TIGIT antagonist antibody is (a) a VH domain comprising an amino acid sequence having at least 95% sequence identity with the amino acid sequence of EVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPRSGL EWL GKTY YRFKWYSDYAVSVKGRITINPDTSKNQFS LQL NSVT PEDTAVFYC TRE STTYDLLAGPF DYWGQGTLVT VSS (SEQ ID NO: 17) or QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPRSGL EWL GKTY YRFKWYSDYAVSVKGRITINPDTSKNQFS LQL NSVT PEDTAVFYC TRE STTYDLLAGPF DYWGQGTLVT VSS (SEQ ID NO: 18); (b) a VL domain comprising an amino acid sequence having at least 95% sequence identity with the amino acid sequence of DIVMTQSPDSLAVSLGERATINCKSSQTVLYSSNNKKYLAYWQ QKPGQPPNLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQ QYYSTPF TF GPGTKVEIK (SEQ ID NO: 19); or (c) the VH domain described in (a) and the VL domain described in (b) The method according to any one of claims 60 to 64, comprising

66. The anti-TIGIT antagonist antibody is (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, The method according to claim 65, comprising

67. The anti-TIGIT antagonist antibody is (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, The method according to claim 66, comprising

68. The anti-TIGIT antagonist antibody is (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, The method according to any one of claims 1 to 62 and 64 to 67, comprising

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

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

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

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

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

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

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

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

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

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

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

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

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

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

84. The method according to claim 83, wherein the anti-PD-L1 antagonist antibody is atezolizumab, MDX-1105, durvalumab, avelumab, SHR-1316, CS1001, enoblituzumab, TQB2450, ZKAB001, LP-002, CX-072, IMC-001, KL-A167, APL-502, cosibelimab, rodaplimab, 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 according to claim 84, wherein the anti-PD-L1 antagonist antibody is atezolizumab.

86. The anti-PD-L1 antagonist antibody has the following HVRs: (a) An HVR-H1 sequence comprising the amino acid sequence of GFTFSDSWI H (SEQ ID NO: 20); (b) An HVR-H2 sequence comprising the amino acid sequence of AWISP YGGSTYYADS VK G (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 RASQDVS TAVA (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 QQYLYH PAT (SEQ ID NO: 25) The method according to claim 83, comprising.

87. The anti-PD-L1 antagonist antibody is (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) The VH domain described in (a) and the VL domain described in (b) The method according to claim 86, comprising.

88. The anti-PD-L1 antagonist antibody is (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, The method according to claim 85, comprising.

89. The anti-PD-L1 antagonist antibody is (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 The method according to claim 88, comprising.

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

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

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

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

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

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

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

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

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

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

100. The method according to claim 99, wherein the anti-PD-1 antagonist antibody is nivolumab, pembrolizumab, MEDI-0680, spartalizumab, semiprimab, BGB-108, prorolimab, camrelizumab, sintilimab, tislelizumab, tripalimumab, dostarlimab, retifanlimab, sasanalimab, pemprimab, CS1003, HLX10, SCT-I10A, zinberelimab, valsiriumab, genolimzumab, BI754091, cetrelimab, YBL-006, BAT1306, HX008, budigalimab, AMG404, CX-188, JTX-4014, 609A, Sym021, LZM009, F520, SG001, AM0001, ENUM 244C8, ENUM 388D4, STI-1110, AK-103 or hAb21.

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

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

103. The method according to 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 the treatment of an individual having cancer, wherein the individual has been determined to have a TAM signature score that exceeds a reference TAM signature score, thereby identifying the individual as an individual who may benefit from treatment with 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-assigned 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 of the TAM signature scores of the reference population.

110. The use according to claim 108 or 109, wherein the reference population is a population of individuals having 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. 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 is detected in the sample from the individual.

114. 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. 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 is detected in the sample from the individual, from which the TAM signature score is determined, and 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 is determined to have a Treg signature score that exceeds a reference Treg signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and 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. 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. 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-assigned Treg signature score.

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

122. The use according to claim 120 or 121, wherein the reference population is a population of individuals having 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 an 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 material.

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

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

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

140. The use according to claim 139, wherein when the PD-L1 positive tumor cell fraction is determined by positive staining using the anti-PD-L1 antibody SP263, it is 50% or more.

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 when the PD-L1 positive tumor cell fraction is determined by positive staining using the anti-PD-L1 antibody 22C3, it is 50% or more.

143. The use according to claim 142, wherein the PD-L1 positive tumor cell fraction is calculated using the pharmDx 22C3 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 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 exceeds a reference TAM signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and wherein the TAM signature score is based on the expression levels of each of C1QC, MSR1, MRC1, VSIG4, SPP1, and MARCO detected in a sample from the individual, the PD-1 axis-binding antagonist and / or anti-TIGIT antagonist antibody.

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. The 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-assigned 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 signature score in the reference population is the median of the TAM signature scores 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 having 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, from which the TAM signature score is determined, and 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 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 exceeds a reference Treg signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody, and the Treg signature score is based on the expression levels of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in a sample from the individual, a PD-1 axis-binding antagonist and / or anti-TIGIT antagonist antibody.

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 the 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 signature score is a pre-assigned Treg signature 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 signature score is the Treg signature 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 signature score in the reference population is the median of the Treg signature scores 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 having 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 signature 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 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.

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 for use 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 an 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 material.

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 archive sample, a fresh sample, or a frozen sample.

179. The 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 is determined to have a PD-L1 positive tumor cell fraction by an 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, and 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 when the PD-L1 positive tumor cell fraction is determined by positive staining using the anti-PD-L1 antibody SP263, it is 50% or more.

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

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

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

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

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

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

188. The anti-TIGIT antagonist antibody can interact with the Fc gamma receptor (FcγR) on myeloid cells and can induce mobilization of CD8+ T cells in the blood and / or expansion of proliferating CD8+ T cells in the tumor bed, the method according to any one of claims 1 to 103, the use according to any one of claims 104 to 145, or the PD-1 axis binding antagonist and / or anti-TIGIT antagonist antibody for use according to any one of claims 146 to 186.

189. A method of identifying an individual having cancer who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting 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 therefrom a tumor-associated macrophage (TAM) signature score, wherein an individual having a TAM signature score above a reference TAM signature score is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

190. A method for selecting a treatment method 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 therefrom a TAM signature score, wherein an individual having a TAM signature score above a reference TAM signature score is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

191. The method according to claim 189 or 190, wherein the individual has a TAM signature score in the sample above 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 exhibiting effector function.

192. A method of treating an individual having 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 exceeds a reference TAM signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, 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; (b) Administering to the individual an effective amount of a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function; A method comprising the steps of.

193. A method of treating an individual having 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 is determined to have a TAM signature score that exceeds a reference TAM signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, and 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 having cancer to a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, the method comprising detecting, in a sample from the individual, during or after administration of the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody exhibiting effector function, 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, 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 each respective reference expression level predicts an individual likely to respond to a treatment comprising the PD-1 axis-binding antagonist and the anti-TIGIT antagonist antibody exhibiting effector function.

195. A method of identifying an individual having cancer who may benefit from a treatment comprising a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting 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 therefrom a regulatory T cell (Treg) signature score, wherein a Treg signature score above 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 exhibiting effector function.

196. A method for selecting a treatment method 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 an individual having a Treg signature score above a reference Treg signature score is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function.

197. The method according to claim 195 or 196, wherein the individual has a Treg signature score in the sample above a reference Treg 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 exhibiting effector function.

198. A method of treating an individual having 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 above a reference Treg signature score, whereby the individual is identified as an individual who may benefit from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, 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; 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. A method comprising.

199. A method of treating an individual having 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 signature score above a reference Treg signature score, whereby the individual is identified as an individual potentially benefiting from treatment with a PD-1 axis-binding antagonist and an anti-TIGIT antagonist antibody exhibiting effector function, and the Treg signature score is based on the expression levels of each of FOXP3, CTLA4, IL10, TNFRSF18, CCR8, IKZF4 and IKZF2 detected in a sample from the individual.

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

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