Diagnostic and therapeutic methods for sarcomatoid kidney cancer

Abstract

To provide diagnostic methods, therapeutic methods, and compositions for the treatment of cancer (e.g., kidney cancer (e.g., renal cell carcinoma (RCC)).SOLUTION: The present invention provides a method of treating an individual having a kidney cancer, the method comprising administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist, wherein the individual has been identified as likely to benefit from the anti-cancer therapy based on having a sarcomatoid kidney cancer.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy, created on October 8, 2019, is entitled 50474-191WO3_Sequence_Listing_10.8.19_ST25 and is 235,579 bytes in size.

[0002] The present invention relates to diagnostic and therapeutic methods for the treatment of cancer (eg, kidney cancer). [Background technology]

[0003] Cancer remains one of the most deadly threats to human health. In the United States, cancer accounts for approximately 1.3 million new cases each year, making it the second leading cause of death after heart disease, accounting for approximately one in four deaths. It has also been predicted that cancer may surpass cardiovascular disease as the leading cause of death within five years. Solid tumors are responsible for the majority of these deaths. While significant advances have been made in the medical treatment of certain cancers, the overall five-year survival rate for all cancers has improved by only about 10% over the past 20 years. Malignant solid tumors, in particular, metastasize and grow rapidly in an uncontrolled manner, making their timely detection and treatment extremely challenging. Renal cell carcinoma (RCC) is the most common type of kidney cancer and has multiple histologic subtypes. Sarcomatoid RCC, which can occur in all histologic subtypes, is characterized in part by features similar to sarcomas, including spindle cells, high cellularity, and cellular atypia. Sarcomatoid RCC is associated with a poor prognosis, with a median survival of approximately 6 months, and a high percentage of sarcomatoid components is associated with a worse outcome. Sarcomatoid RCC is typically difficult to treat and is considered to be an aggressive form of RCC.

[0004] Human studies using immune checkpoint inhibitors have demonstrated the potential to harness the immune system to suppress and eradicate tumor growth. The programmed death 1 (PD-1) receptor and its ligand, programmed death-ligand 1 (PD-L1), are immune checkpoint proteins involved in suppressing immune system responses during chronic infection, pregnancy, tissue allografts, autoimmune diseases, and cancer. PD-L1 regulates immune responses by binding to the inhibitory receptor PD-1, which is expressed on the surface of T cells, B cells, and monocytes. PD-L1 also negatively regulates T cell function through its interaction with another receptor, B7-1. Formation of the PD-L1 / PD-1 and PD-L1 / B7-1 complexes negatively regulates T cell receptor signaling, subsequently downregulating T cell activation and suppressing antitumor immune activity.

[0005] Despite significant advances in the treatment of cancer (eg, kidney cancer), improved diagnostic methods and cancer therapies remain needed. Summary of the Invention

[0006] The present invention provides diagnostic and therapeutic methods and compositions for treating individuals with cancer (e.g., kidney cancer (e.g., renal cell carcinoma (RCC))), including sarcomatoid cancer (e.g., sarcomatoid renal cancer (e.g., locally advanced or metastatic sarcomatoid RCC)).

[0007] In one aspect, the invention features a method of treating an individual with a sarcomatoid cancer (e.g., a sarcomatoid renal cancer (e.g., a sarcomatoid RCC, including locally advanced or metastatic sarcomatoid RCC)), comprising administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist. In some embodiments, the individual has not previously received treatment for the sarcomatoid cancer.

[0008] In another aspect, the invention features a method of treating an individual with a cancer (e.g., kidney cancer (e.g., RCC, including locally advanced or metastatic RCC)) with a low or intermediate Memorial Sloan Kettering Cancer Center (MSKCC) risk score, the method comprising administering to the individual an anti-cancer therapy comprising an effective amount of a VEGF antagonist and a PD-L1 axis-binding antagonist. In some embodiments, the individual has not previously received treatment for the cancer.

[0009] In another aspect, the invention features a method of treating an individual having renal cancer, the method including administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the individual having sarcomatoid renal cancer and having been identified as likely to benefit from the anti-cancer therapy.

[0010] In another aspect, the invention features a method of treating an individual with kidney cancer, the method including: (a) determining whether the individual has sarcomatoid kidney cancer, wherein the presence of sarcomatoid kidney cancer indicates that the individual may benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist; and (b) administering to the individual an effective amount of an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the presence of sarcomatoid kidney cancer.

[0011] In another aspect, the invention features a method for identifying an individual with renal cancer who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the method comprising determining whether the individual has sarcomatoid renal cancer, wherein the presence of sarcomatoid renal cancer identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0012] In another aspect, the invention features a method for selecting a treatment for an individual with renal cancer, the method comprising: (a) determining whether the individual has sarcomatoid renal cancer, where the presence of sarcomatoid renal cancer identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist; and (b) selecting an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the presence of sarcomatoid renal cancer. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0013] In another aspect, the invention features a pharmaceutical composition comprising a PD-L1 axis-binding antagonist for use in treating an individual with a sarcomatoid cancer (e.g., sarcomatoid renal cancer (e.g., sarcomatoid RCC, including locally advanced or metastatic sarcomatoid RCC)), wherein the treatment comprises administration of a PD-L1 axis-binding antagonist in combination with a VEGF antagonist. In some embodiments, the individual has not previously received treatment for the sarcomatoid cancer.

[0014] In another aspect, the invention provides use of a PD-L1 axis-binding antagonist in the manufacture of a medicament for treating an individual with a sarcomatoid cancer (e.g., sarcomatoid renal cancer (e.g., sarcomatoid RCC, including locally advanced or metastatic sarcomatoid RCC)), wherein the treatment comprises administering a PD-L1 axis-binding antagonist in combination with a VEGF antagonist. In some embodiments, the individual has not previously received treatment for the sarcomatoid cancer.

[0015] In another aspect, the invention features a pharmaceutical composition including a PD-L1 axis binding antagonist for use in treating an individual having renal cancer, the treatment comprising administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist, wherein the individual has been identified as likely to benefit from the anti-cancer treatment based on the individual having sarcomatoid renal cancer.

[0016] In another aspect, the invention provides the use of a PD-L1 axis binding antagonist in the manufacture of a medicament for the treatment of an individual having renal cancer, wherein the treatment comprises administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist, and the individual is identified as being likely to benefit from the anti-cancer treatment by virtue of having sarcomatoid renal cancer.

[0017] In some embodiments of any of the aforementioned aspects, the presence of sarcomatoid renal cancer is assessed by histological analysis of a sample obtained from the individual. In some embodiments, the renal cancer is sarcomatoid if a tumor sample from the individual contains foci (or foci) of any component of high-grade malignant spindle cells relative to the overall tumor area. In some embodiments, the spindle cells exhibit moderate to marked atypia and / or resemble any form of sarcoma. In some embodiments, the spindle cells show evidence of epithelial differentiation, as assessed by immunohistochemical positivity for keratin or epithelial membrane antigen (EMA). In some embodiments, the renal cancer is renal cell carcinoma, and the tumor sample has epithelial differentiation with concurrent areas of renal cell carcinoma.

[0018] In some embodiments of any of the foregoing aspects, the benefit relates to improved progression-free survival (PFS), overall survival (OS), overall response rate (ORR), complete response (CR) rate, or disease progression-free rate (DFR). In some embodiments, the benefit relates to improved PFS. In some embodiments, the benefit relates to improved OS. In some embodiments, the benefit relates to improved ORR. In some embodiments, the benefit relates to improved CR rate. In some embodiments, the benefit relates to improved DFR. In some embodiments, the DFR is determined as the time from initiation of treatment to the individual's first increase of 2 or more points above baseline on the MD Anderson Symptom Assessment Index (MDASI) Disability Scale.

[0019] In some embodiments of any of the aforementioned aspects, the individual has a low or intermediate Memorial Sloan Kettering Cancer Center (MSKCC) risk score.

[0020] In another aspect, the invention features a method of treating an individual with kidney cancer, the method including administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, wherein the individual has been identified as likely to benefit from the anti-cancer therapy based on having a low or intermediate MSKCC risk score.

[0021] In another aspect, the invention features a method of treating an individual with kidney cancer, the method including: (a) determining the individual's MSKCC risk score, where a low or moderate MSKCC risk score indicates the individual is likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist; and (b) administering to the individual an effective amount of an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the individual having a low or moderate MSKCC risk score.

[0022] In another aspect, the invention features a method of identifying an individual having kidney cancer who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the method including determining the individual's MSKCC risk score, wherein a low or intermediate MSKCC risk score identifies an individual as likely to benefit from an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0023] In another aspect, the invention features a method for selecting a treatment for an individual with kidney cancer, the method including: (a) determining the individual's MSKCC risk score, wherein a low or moderate MSKCC risk score identifies the individual as likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist; and (b) selecting the anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the individual having a low or moderate MSKCC risk score.

[0024] In another aspect, the invention features a pharmaceutical composition including a PD-L1 axis binding antagonist for use in treating an individual having renal cancer, the treatment comprising administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist, and the individual is identified as likely to benefit from the anti-cancer treatment based on having a low or intermediate MSKCC risk score.

[0025] In another aspect, the invention provides use of a PD-L1 axis binding antagonist in the manufacture of a medicament for the treatment of an individual with kidney cancer, wherein the treatment comprises administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist, and the individual is identified as likely to benefit from the anti-cancer treatment based on having a low or intermediate MSKCC risk score.

[0026] In another aspect, the invention features a pharmaceutical composition comprising a PD-L1 axis binding antagonist for use in treating an individual with a cancer with a low or intermediate MSKCC risk score (e.g., kidney cancer (e.g., RCC, including locally advanced or metastatic RCC)), wherein the treatment comprises administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist. In some embodiments, the individual has not received previous treatment for the cancer.

[0027] In another aspect, the invention provides use of a PD-L1 axis binding antagonist in the manufacture of a medicament for treating an individual with a cancer with a low or intermediate MSKCC risk score (e.g., kidney cancer (e.g., RCC, including locally advanced or metastatic RCC)), wherein the treatment comprises administering a PD-L1 axis binding antagonist in combination with a VEGF antagonist. In some embodiments, the individual has not previously received treatment for the cancer.

[0028] In some embodiments of any of the aforementioned aspects, an individual has a high MSKCC risk score if the individual has three or more of the following characteristics: (i) time from nephrectomy to systemic treatment less than 1 year, lack of nephrectomy, or initial diagnosis of metastatic disease; (ii) a hemoglobin level below the lower limit of normal (LLN) (in some cases, the normal range for hemoglobin is 13.5-17.5 g / dL for men and 12-15.5 g / dL for women); (iii) a corrected serum calcium level greater than 10 mg / dL (in some cases, the corrected serum calcium level is serum calcium level (mg / dL) + 0.8(4-serum albumin (g / dL))); iv) a serum lactate dehydrogenase (LDH) level greater than 1.5 times the upper limit of normal (ULN) (in some cases, the ULN is 140 U / L); and / or (v) a Karnofsky Performance Status (KPS) score less than 80.

[0029] In some embodiments of any of the foregoing aspects, an individual has a moderate MSKCC risk score if the individual has one or two of the following characteristics: (i) time from nephrectomy to systemic treatment less than 1 year, lack of nephrectomy, or initial diagnosis of metastatic disease; (ii) a hemoglobin level below the LLN (optionally, the normal range for hemoglobin is 13.5-17.5 g / dL for men and 12-15.5 g / dL for women); (iii) a corrected serum calcium level greater than 10 mg / dL (optionally, the corrected serum calcium level is serum calcium level (mg / dL) + 0.8(4-serum albumin (g / dL))); (iv) a serum LDH level greater than 1.5 times the ULN (optionally, the ULN is 140 U / L); and / or (v) a KPS score less than 80.

[0030] In some embodiments of any of the foregoing aspects, the benefit relates to improved progression-free survival (PFS), overall survival (OS), overall response rate (ORR), complete response (CR) rate, or disease progression-free rate (DFR). In some embodiments, the benefit relates to improved PFS. In some embodiments, the benefit relates to improved OS. In some embodiments, the benefit relates to improved ORR. In some embodiments, the benefit relates to improved CR rate. In some embodiments, the benefit relates to improved DFR. In some embodiments, the DFR is determined as the time from initiation of treatment to the individual's first increase of 2 or more points above baseline on the MD Anderson Symptom Assessment Index (MDASI) Disability Scale.

[0031] In some embodiments of any of the aforementioned aspects, the individual has sarcomatoid renal cancer.

[0032] In some embodiments of any of the aforementioned aspects, the method further includes determining the expression level of one or more of the following genes in a sample from the individual: CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2; VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34; or IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2.

[0033] In some embodiments of any of the foregoing aspects, (i) an expression level of one or more of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample that is equal to or greater than a reference expression level for the one or more genes, or (ii) an expression level of one or more of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34, or IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample that is less than a reference expression level for the one or more genes, identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0034] In some embodiments of any of the aforementioned aspects, the expression level of one or more of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample is determined to be equal to or greater than a reference level for the one or more genes. In some embodiments, the expression levels of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or all twenty of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample are determined to be equal to or greater than a reference level for the one or more genes. In some embodiments, the expression level of one or more of CD8A, EOMES, PRF1, IFNG, or PD-L1 in the sample is determined to be equal to or greater than a reference level for the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 in the sample are determined to be equal to or greater than a reference level of CD8A, EOMES, PRF1, IFNG, and PD-L1.

[0035] In some embodiments of any of the foregoing aspects, the expression level of one or more of IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample is determined to be equal to or greater than the reference level of the one or more genes. In some embodiments, the expression levels of at least one, at least two, at least three, at least four, at least five, or all six of IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample are determined to be equal to or greater than the reference level of the one or more genes. In some embodiments, the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, and PTGS2 in the sample are determined to be equal to or greater than the reference level of IL6, CXCL1, CXCL2, CXCL3, CXCL8, and PTGS2. In some embodiments, the expression level of PD-L1 in the sample is determined to be equal to or greater than a reference expression level of PD-L1, and the expression level of one or more additional genes selected from the group consisting of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample is determined to be equal to or greater than a reference expression level of the one or more additional genes.

[0036] In some embodiments of any of the aforementioned aspects, the expression level of one or more of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 in the sample is determined to be less than the reference level of the one or more genes.In some embodiments, the expression level of at least one, at least two, at least three, at least four, at least five, at least six, or all seven of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 in the sample is determined to be less than the reference level of the one or more genes.In some embodiments, the expression level of one or more of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, or CD34 in the sample is determined to be less than the reference level of the one or more genes. In some embodiments, the expression levels of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 in the sample are determined to be less than the reference levels of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34.

[0037] In some embodiments of any of the foregoing aspects, the expression level of one or more of IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample is determined to be less than the reference level of the one or more genes. In some embodiments, the expression levels of at least one, at least two, at least three, at least four, at least five, or all six of IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample are determined to be less than the reference level of the one or more genes. In some embodiments, the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, and PTGS2 in the sample are determined to be less than the reference levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, and PTGS2.

[0038] In some embodiments of any of the above-mentioned aspects, the reference level of one or more genes is determined from a group of individuals with renal cancer.In some embodiments, the reference level of one or more genes is the median expression level determined in a group of patients with renal cancer.In some embodiments, the reference level is the median Z-score of the normalized expression level of one or more genes.

[0039] In some embodiments of any of the above aspects, the expression level is a nucleic acid expression level. 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, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

[0040] In other embodiments of any of the foregoing aspects, the expression level is a protein expression level. In some embodiments, the protein expression level is determined by immunohistochemistry (IHC), Western blotting, enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, immunofluorescence, radioimmunoassay, or mass spectrometry.

[0041] In some embodiments of any of the aforementioned aspects, the sample is a tissue sample, a cell sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof. In some embodiments, the tissue sample is a tumor tissue sample. In some embodiments, the tumor tissue sample comprises tumor cells, tumor-infiltrating immune cells, stromal cells, or a combination thereof. In some embodiments, the tumor tissue sample is a formalin-fixed and paraffin-embedded (FFPE) sample, an archival sample, a fresh sample, or a frozen sample.

[0042] In some embodiments of any of the aforementioned aspects, the individual has not been previously treated for kidney cancer.

[0043] In some embodiments of any of the foregoing aspects, the kidney cancer is renal cell carcinoma (RCC). In some embodiments, the RCC is clear cell RCC. In some embodiments, the RCC is locally advanced or metastatic RCC (mRCC).

[0044] In some embodiments of any of the foregoing aspects, a tumor sample obtained from a patient is determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise about 1% or more of the tumor sample. In some embodiments, the tumor sample is determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise about 1% or more but less than 5% of the tumor sample. In some embodiments, the tumor sample is determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise about 5% or more but less than 10% of the tumor sample. In some embodiments, the tumor sample obtained from a patient is determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise about 10% or more of the tumor sample.

[0045] In other embodiments of any of the aforementioned aspects, a tumor sample obtained from the patient has been determined to have detectable expression levels of PD-L1 in tumor-infiltrating immune cells that comprise less than 1% of the tumor sample.

[0046] In some embodiments of any of the foregoing aspects, the VEGF antagonist is an anti-VEGF antibody or a VEGF receptor (VEGFR) inhibitor. In some embodiments, the VEGF antagonist is an anti-VEGF antibody. In some embodiments, the anti-VEGF antibody is bevacizumab. In some embodiments, the VEGF antagonist is a VEGFR inhibitor. In some embodiments, the VEGFR inhibitor is a multi-targeted tyrosine kinase inhibitor. In some embodiments, the multi-targeted tyrosine kinase inhibitor is sunitinib, axitinib, pazopanib, or cabozantinib. In some embodiments, the multi-targeted tyrosine kinase inhibitor is sunitinib.

[0047] In some embodiments of any of the aforementioned aspects, the PD-L1 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. In some embodiments, the PD-L1 axis binding antagonist is a PD-L1 binding antagonist. 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. In some embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to B7-1. In some embodiments, the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7-1. In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody is selected from the group consisting of MPDL3280A (atezolizumab), YW243.55.S70, MDX-1105, MEDI4736 (durvalumab), and MSB0010718C (avelumab). In some embodiments, the anti-PD-L1 antibody comprises the following hypervariable regions (HVRs): (a) the HVR-H1 sequence of GFTFSDSWIH (SEQ ID NO: 63); (b) the HVR-H2 sequence of AWISPYGGSTYYADSVKG (SEQ ID NO: 64); (c) the HVR-H3 sequence of RHWPGGFDY (SEQ ID NO: 65); (d) the HVR-L1 sequence of RASQDVSTAVA (SEQ ID NO: 66); (e) the HVR-L2 sequence of SASFLYS (SEQ ID NO: 67); and (f) the HVR-L3 sequence of QQYLYHPAT (SEQ ID NO: 68).In some embodiments, the anti-PD-L1 antibody comprises: (a) a heavy chain variable (VH) domain comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 69); (b) a light chain variable (VL) domain comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 70); or (c) a VH domain similar to (a) and a VL domain similar to (b). In some embodiments, the anti-PD-L1 antibody comprises: (a) a VH domain comprising an amino acid sequence that has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 69; (b) a VL domain comprising an amino acid sequence that has at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain similar to (a) and a VL domain similar to (b). In some embodiments, the anti-PD-L1 antibody comprises: (a) a VH domain comprising an amino acid sequence that has at least 96% sequence identity to the amino acid sequence of SEQ ID NO: 69; (b) a VL domain comprising an amino acid sequence that has at least 96% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain similar to (a) and a VL domain similar to (b). In some embodiments, the anti-PD-L1 antibody comprises: (a) a VH domain comprising an amino acid sequence that has at least 97% sequence identity to the amino acid sequence of SEQ ID NO: 69; (b) a VL domain comprising an amino acid sequence that has at least 97% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain similar to (a) and a VL domain similar to (b).In some embodiments, the anti-PD-L1 antibody comprises: (a) a VH domain comprising an amino acid sequence that has at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 69; (b) a VL domain comprising an amino acid sequence that has at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain similar to (a) and a VL domain similar to (b). In some embodiments, the anti-PD-L1 antibody comprises: (a) a VH domain comprising an amino acid sequence that has at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 69; (b) a VL domain comprising an amino acid sequence that has at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain similar to (a) and a VL domain similar to (b). In some embodiments, the anti-PD-L1 antibody comprises: (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 69; (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain similar to (a) and a VL domain similar to (b). In some embodiments, the anti-PD-L1 antibody comprises: (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 69; and (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 70. In some embodiments, the anti-PD-L1 antibody is atezolizumab.

[0048] In some embodiments of any of the foregoing aspects, the PD-L1 axis-binding antagonist is atezolizumab and the VEGF antagonist is bevacizumab. In some embodiments, atezolizumab is administered intravenously at a dose of about 1200 mg every three weeks. In some embodiments, bevacizumab is administered intravenously at a dose of about 15 mg / kg every three weeks.

[0049] In some embodiments of any of the aforementioned aspects, the method further comprises administering to the individual an additional therapeutic agent. In some embodiments, the additional therapeutic agent is selected from the group consisting of an immunotherapeutic agent, a cytotoxic agent, a growth inhibitory agent, a radiotherapeutic agent, an anti-angiogenic agent, and combinations thereof. In some embodiments, the individual is human. [Brief explanation of the drawings]

[0050] The application file contains at least one drawing executed in color. Copies of this patent or this patent application with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. [Figure 1] Schematic diagram showing the IMmotion151 trial design. Primary endpoints were progression-free survival (PFS) (investigator-assessed PFS per RECIST v1.1) in the PD-L1+ subgroup and overall survival (OS) in the intention-to-treat (ITT) population. Exploratory endpoints included validation of gene signatures from the IMmotion150 trial and their association with PFS, as well as biomarker characterization in Memorial Sloan Kettering Cancer Center (MSKCC) risk subgroups and sarcomatoid tumors. a ≥1% IC: 40% prevalence using the SP142 immunohistochemistry (IHC) assay. b No atezolizumab or bevacizumab dose reductions. [Figure 2]

[0023] Figure 1 is a series of graphs showing Kaplan-Meier curves illustrating the rate of PFS in the PD-L1+ subgroup (left panel) and the ITT population (right panel) of patients treated with atezolizumab and bevacizumab ("atezo+bevac") or sunitinib. The table shows the median PFS (months) and 95% confidence interval (95% CI). PFS was assessed by the investigator. Minimum follow-up, 12 months. Median follow-up, 16 months (PD-L1+) and 15 months (ITT). aPFS analysis crossed the pre-specified P-value boundary of α = 0.04. [Figure 3] FIG. 1 is a schematic diagram showing the gene signature analysis scheme for the IMmotion151 study. [Figure 4] FIG. 1 is a heatmap showing that the IMmotion151 transcriptome map confirmed the biological subgroups identified in the IMmotion150 study. [Figure 5]This is a series of graphs showing Kaplan-Meier curves showing the rate of PFS in the angiogenesis (Angio)-low (left panel) or Angio-high (right panel) subgroups of patients treated with Atezo+Beva or sunitinib. Atezo+Beva improved PFS compared with sunitinib in the Angio-low subgroup. The table shows the hazard ratios (HRs) and 95% CI. [Figure 6] A series of graphs showing Kaplan-Meier curves showing the proportion of PFS in patients treated with sunitinib (left panel) or atezoh plus bevastatin (right panel). Sunitinib demonstrated improved PFS in the Angio-high versus Angio-low subgroups. The table shows hazard ratios (HRs) and 95% CIs. [Figure 7] This is a series of graphs showing Kaplan-Meier curves showing the rate of PFS in T effector (Teff)-low (left panel) or Teff-high (right panel) subgroups of patients treated with Atezo+Beva or sunitinib. Atezo+Beva improved PFS compared with sunitinib in the Teff-high subgroup. The table shows the hazard ratios (HRs) and 95% CI. [Figure 8A] Graph showing the results of subgroup PFS analysis in PD-L1+ and all evaluable patients (biomarker-evaluable population). [Figure 8B] Graph showing Kaplan-Meier curves showing the proportion of PFS in patients treated with sunitinib or atezoheptone plus bevacizumab. Atezoheptone plus bevacizumab treatment showed improved PFS in sarcomatoid tumors. Table shows HR and 95% CI. [Figure 9A] Figure 9A is a series of graphs showing the expression of the Angio gene signature (Figure 9A), the Teff gene signature (Figure 9B), and PD-L1 (Figure 9C) in sarcomatoid and non-sarcomatoid subgroups. Sarcomatoid tumors had lower expression of the Angio gene signature and higher expression of PD-L1. [Figure 9B]Figure 9A is a series of graphs showing the expression of the Angio gene signature (Figure 9A), the Teff gene signature (Figure 9B), and PD-L1 (Figure 9C) in sarcomatoid and non-sarcomatoid subgroups. Sarcomatoid tumors had lower expression of the Angio gene signature and higher expression of PD-L1. [Figure 9C] Figure 9A is a series of graphs showing the expression of the Angio gene signature (Figure 9A), the Teff gene signature (Figure 9B), and PD-L1 (Figure 9C) in sarcomatoid and non-sarcomatoid subgroups. Sarcomatoid tumors had lower expression of the Angio gene signature and higher expression of PD-L1. [Figure 10A] 10A-10C are a series of graphs showing the expression of the Angio signature (FIG. 10A), Teff signature (FIG. 10B), and PD-L1 (FIG. 10C) in low or intermediate / high MSKCC risk subgroups. Expression of the Angio gene signature was higher in the low MSKCC risk group. [Figure 10B] 10A-10C are a series of graphs showing the expression of the Angio signature (FIG. 10A), Teff signature (FIG. 10B), and PD-L1 (FIG. 10C) in low or intermediate / high MSKCC risk subgroups. Expression of the Angio gene signature was higher in the low MSKCC risk group. [Figure 10C] 10A-10C are a series of graphs showing the expression of the Angio signature (FIG. 10A), Teff signature (FIG. 10B), and PD-L1 (FIG. 10C) in low or intermediate / high MSKCC risk subgroups. Expression of the Angio gene signature was higher in the low MSKCC risk group. [Figure 11A]

[0033] Figure 11A is a series of graphs showing Kaplan-Meier curves illustrating the rate of PFS for patients treated with ATEZO+BEVA or sunitinib versus all patients with sarcomatoid tumors ("All Sarc") (Figure 11A) or PD-L1+ tumors ("PD-L1+ Sarc") (Figure 11B). Patients with sarcomatoid histology in the ATEZO+BEVA arm had a longer median PFS than patients in the sunitinib arm, regardless of PD-L1+ status. [Figure 11B]

[0033] Figure 11A is a series of graphs showing Kaplan-Meier curves illustrating the rate of PFS for patients treated with ATEZO+BEVA or sunitinib versus all patients with sarcomatoid tumors ("All Sarc") (Figure 11A) or PD-L1+ tumors ("PD-L1+ Sarc") (Figure 11B). Patients with sarcomatoid histology in the ATEZO+BEVA arm had a longer median PFS than patients in the sunitinib arm, regardless of PD-L1+ status. [Figure 12A]

[0023] Figure 12A is a series of graphs showing Kaplan-Meier curves illustrating the rate of overall survival (OS) for patients treated with ATEZO+BEVA or sunitinib versus all patients with sarcomatoid tumors ("All Sarc") (Figure 12A) or PD-L1+ tumors ("PD-L1+ Sarc") (Figure 12B). OS was increased in patients with sarcomatoid histology treated with ATEZO+BEVA versus patients treated with sunitinib, regardless of PD-L1+ status. [Figure 12B]

[0023] Figure 12A is a series of graphs showing Kaplan-Meier curves illustrating the rate of overall survival (OS) for patients treated with ATEZO+BEVA or sunitinib versus all patients with sarcomatoid tumors ("All Sarc") (Figure 12A) or PD-L1+ tumors ("PD-L1+ Sarc") (Figure 12B). OS was increased in patients with sarcomatoid histology treated with ATEZO+BEVA versus patients treated with sunitinib, regardless of PD-L1+ status. [Figure 13]Figure 1 shows time to deteriorationa, symptom interference with daily functionb, for all patients with sarcomatoid tumors. DFR, deterioration-free rate. a Time to clinically meaningful deterioration was prespecified as the time from randomization to the patient's first increase of 2 or more points above baseline on the MD Anderson Symptom Assessment Inventory (MDASI) Interference scale (range, 0-10) (see, e.g., Mendoza et al., Clin. Breast Cancer 13:325-334, 2013; Jones et al., Clin. Genitourin. Cancer 12:41-49, 2014; and Shi et al., Pain 158:1108-1112, 2017). b Measures of daily functioning included work, general activities, walking, relationships with others, enjoyment of life, and mood. DETAILED DESCRIPTION OF THE INVENTION

[0051] The present invention provides diagnostic methods, therapeutic methods and uses, and compositions for the treatment of cancer, including sarcomatoid cancer (e.g., kidney cancer (e.g., renal cell carcinoma (RCC)). The present invention provides methods for the diagnosis of sarcomatoid cancer (e.g., sarcomatoid kidney cancer, such as sarcomatoid RCC) and / or an individual's Memorial Sloan Kettering Cancer Center (MSKCC) risk score, and for determining whether the individual has received a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab) or a VEGFR inhibitor (e.g., VEGF inhibitor). For example, identifying whether an individual is likely to respond to treatment including a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)) and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab (MPDL3280A)) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), selecting a therapy for treating the individual, a VEGF antagonist, and The present invention is based, at least in part, on the discovery that the PD-L1 axis-binding antagonists can be used as biomarkers (e.g., predictive biomarkers) in methods for optimizing the therapeutic efficacy of treatments comprising VEGF antagonists and PD-L1 axis-binding antagonists, and / or monitoring an individual's response to treatments comprising VEGF antagonists and PD-L1 axis-binding antagonists. The present invention also relates to the use of VEGF antagonists (e.g., anti-VEGF antibodies (e.g., bevacizumab) or VEGFR inhibitors (e.g., multi-targeted tyrosine kinase inhibitors (TKIs)). Methods are provided for treating an individual having cancer (e.g., kidney cancer (e.g., renal cell carcinoma (RCC)) by administering an anti-cancer therapy comprising a PD-L1 axis-binding antagonist (e.g., a PD-L1 enzyme inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)) and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab (MPDL3280A)) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

[0052] I. Definition Aspects and embodiments of the invention described herein should be understood to include aspects and embodiments "comprising," "consisting of," and "consisting essentially of." As used herein, the singular forms "a," "an," and "the" include plurals unless otherwise indicated.

[0053] As used herein, the term "about" refers to a normal range of error for each value, which would be readily understood by one of ordinary skill in the art. Reference herein to "about" a value or parameter includes (and describes) embodiments that are directed to that value or parameter itself.

[0054] As used herein, the terms "individual," "patient," or "subject" are used interchangeably and refer to any single animal, more preferably a mammal (including, for example, cats, dogs, horses, rabbits, zoo animals, cows, pigs, sheep, and non-human animals such as non-human primates), for which treatment is desired. In certain embodiments, the patient herein is a human. The patient may also be a "cancer patient," i.e., a patient suffering from or at risk of suffering from cancer (e.g., kidney cancer (e.g., RCC)), or one or more symptoms of cancer.

[0055] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by uncontrolled cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More specific examples of such cancers include, but are not limited to, kidney or renal cancer (e.g., renal cell carcinoma (RCC)), lung cancer (including small cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous cell carcinoma of the lung), bladder cancer (e.g., urothelial bladder cancer (UBC), muscle-invasive bladder cancer (MIBC), and BCG-refractory non-muscle-invasive bladder cancer (NMIBC)), cancers of the urinary tract, breast cancer (e.g., HER2+ breast cancer and triple-negative breast cancer (TNBC), which are estrogens, HER2+ breast cancer, ... Gen-receptor (ER-), progesterone receptor (PR-), and HER2 (HER2-) negative), prostate cancer such as castration-resistant prostate cancer (CRPC), cancer of the peritoneum, hepatocellular carcinoma, gastric cancer or stomach cancer (including gastrointestinal cancer and gastrointestinal stromal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer (e.g., hepatocellular carcinoma (HCC)), liver cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, Penile cancer, melanoma (including superficial spreading melanoma, lentigo maligna melanoma, acral lentiginous melanoma, and nodular melanoma), multiple myeloma and B-cell lymphoma (low-grade / follicular non-Hodgkin's 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, bulky disease NHL, mantle cell lymphoma, AIDS-related lymphoma, and Waldenstrom's macrocytoma) In some embodiments, the cancer is renal cancer, including leukemia, ...In certain embodiments, the kidney cancer is RCC (e.g., advanced or metastatic RCC (mRCC), including previously untreated RCC). In some embodiments, the kidney cancer is sarcomatoid kidney cancer (e.g., sarcomatoid RCC (e.g., sarcomatoid advanced or mRCC)).

[0056] "Early stage cancer" or "early stage tumor" means a cancer that is neither invasive nor metastatic and is classified as stage 0, stage I, or stage II cancer.

[0057] An "advanced" cancer is one that has spread outside its original site or organ, either by local invasion or metastasis.

[0058] A "refractory" cancer is one that progresses despite anti-tumor agents, such as chemotherapy, being administered to the cancer patient. An example of a refractory cancer is one that is platinum-refractory.

[0059] A "recurrent" cancer is one that has regrown, either at the original site or at a distant site, after responding to initial treatment.

[0060] The terms "cell proliferative disorder" and "proliferative disorder" refer to disorders associated with some degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer.

[0061] The term "tumor," as used herein, refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues.

[0062] The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive when referred to herein.

[0063] A "disorder" is any condition that would benefit from treatment, including, but not limited to, chronic and acute disorders or diseases, including pathological conditions that predispose a mammal to the disorder in question.

[0064] The term "sarcomatoid" refers to a cancer (e.g., a kidney cancer (e.g., RCC)) characterized by a sarcomatoid morphology, e.g., as assessed by histology. Sarcomatoid kidney cancer (e.g., sarcomatoid RCC) is associated with aggressive behavior and a poor prognosis. In some embodiments, sarcomatoid kidney cancer comprises or consists of atypical spindle cells and / or resembles the morphology of any sarcoma. See, e.g., El Mouallem et al., Urol. Oncol. 36:265-271, 2018, incorporated herein by reference in its entirety. Sarcomatoid RCC can occur in any subtype of RCC, including clear cell RCC, chromophobe RCC, collecting duct carcinoma, renal medullary carcinoma, fumarate hydratase (FH)-deficient RCC, and succinate dehydrogenase (SDH)-deficient RCC. The incidence of sarcomatoid RCC varies by subtype but is usually higher in clear cell RCC (approximately 5-8%) and chromophobe RCC (approximately 8-10%). The histology of the sarcomatoid component can vary and may include fibrosarcomatoid, pleomorphic undifferentiated sarcomatoid, or other heterogeneous sarcomatoid patterns (e.g., osteosarcoma, chondrosarcoma, or rhabdomyosarcoma). Necrosis is usually present in the majority (approximately 90%) of cases. In some embodiments, there is no minimum amount or percentage of sarcomatoid differentiation required for an individual's kidney cancer to be classified as sarcomatoid. Sarcomatoid RCC can be evaluated as described in Example 1. In other embodiments, sarcomatoid RCC can be characterized as described by the 2012 International Society of Urological Pathology (ISUP) Vancouver Consensus (see Srigley et al., Am. J. Surg. Pathol. 37:1469-89, 2013, incorporated herein by reference in its entirety).

[0065] The term "Memorial Sloan Kettering Cancer Center (MSKCC) risk score" refers to a scoring system based on a set of prognostic factors associated with survival in patients with kidney cancer (e.g., RCC, e.g., mRCC). See, e.g., Motzer et al., J. Clin. Oncol. 17(8):2530-2540, 1999 and Motzer et al., J. Clin. Oncol. 20(1):289-296, 2002, which are incorporated by reference in their entireties. In some embodiments, the MSKCC risk score can be calculated based on the following factors, as described in Example 1: (i) time from nephrectomy to treatment (e.g., systemic treatment) less than 1 year, lack of nephrectomy, or initial diagnosis of metastatic disease; (ii) hemoglobin level below the lower limit of normal (LLN) (in some cases, the normal range for hemoglobin is 13.5-17.5 g / dL for men and 12-15.5 g / dL for women); (iii) corrected serum calcium level greater than 10 mg / dL (in some cases, the corrected serum calcium level is serum calcium level (mg / dL) + 0.8 (4 - serum albumin (g / dL))); (iv) serum lactate dehydrogenase (LDH) level greater than 1.5 times the upper limit of normal (ULN) (in some cases, the ULN is 140 U / L); and / or (v) Karnofsky Performance Status (KPS) score less than 80. In some embodiments, if an individual does not have the above characteristics, the individual has a low MSKCC risk score. In some embodiments, if an individual has one or two of the above characteristics, the individual has a moderate MSKCC risk score. In some embodiments, if an individual has three or more of the above characteristics, the individual has a high MSKCC risk score. In some embodiments, an individual's MSKCC risk score can be used to identify whether the individual will benefit from anti-cancer treatment, for example, anti-cancer treatment including a VEGF antagonist (e.g., an anti-VEGF antibody such as bevacizumab) and a PD-L1 axis binding antagonist (e.g., an anti-PD-L1 antibody such as atezolizumab).

[0066] The term "detection" includes any means of detection, including direct or indirect detection.

[0067] The term "sample," as used herein, refers to a composition obtained or derived from a patient and / or individual of interest, containing cells and / or other molecular entities to be characterized and / or identified, for example, based on physical, biochemical, chemical, and / or physiological characteristics. Samples include, but are not limited to, tissue samples, primary or cultured cells or cell lines, cell supernatants, cell lysates, platelets, serum, plasma, vitreous fluid, lymphatic fluid, synovial fluid, follicular fluid, semen, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, tumor lysates, and tissue culture media, tissue extracts, e.g., homogenized tissue, tumor tissue, cell extracts, and combinations thereof.

[0068] As used herein, the terms "cell," "cell line," and "cell culture" are used interchangeably, and all such designations include progeny. Thus, the terms "transformant" and "transformed cell" include the primary subject cell and cultures derived therefrom, regardless of the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content due to deliberate or unintended mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where a separate designation is intended, it will be clear from the context.

[0069] The terms "biomarker" and "marker" are used interchangeably herein to refer to DNA, RNA, protein, carbohydrate, glycolipid, cell-based molecular marker, histological or morphological marker (e.g., sarcomatoid morphology), or risk score (e.g., MSKCC risk score), the expression, presence, and / or level of which in a patient sample can be detected by standard methods (or methods disclosed herein). Such markers include the presence of sarcomatoid kidney cancer (e.g., sarcomatoid RCC) and / or an individual's MSKCC risk score (e.g., a low or intermediate MSKCC risk score). Such biomarkers include, but are not limited to, CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, TAP2, VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, CD34, IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and / or S100A9. The presence, expression, and / or level of such biomarkers may be determined to be higher or lower in a sample obtained from a patient who is sensitive or responsive to treatment (e.g., treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, or treatment with a multi-targeted tyrosine kinase inhibitor) than a reference level (including, for example, the median expression level of the biomarker in samples from a group / population of patients, e.g., patients who have cancer and are being tested for responsiveness to treatment; the median expression level of the biomarker in samples from a group / population of patients, e.g., patients who have cancer and are identified as not responding to treatment; levels in a sample previously obtained from an individual; or levels in a sample from a patient in an early tumor state, and who may now be metastatic, and who has previously received treatment (e.g., anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, or treatment with a multi-targeted tyrosine kinase inhibitor).

[0070] The term "CD8A," as used herein, unless otherwise indicated, refers to any native CD8A from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CD8A and any form of CD8A that results from processing in cells. The term also encompasses naturally occurring variants of CD8A, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CD8A is set forth in SEQ ID NO: 1. The amino acid sequence of an exemplary protein encoded by human CD8A is set forth in SEQ ID NO: 2.

[0071] As used herein, the term "EOMES," unless otherwise indicated, refers to any naturally occurring EOMES (eomesodermin) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed EOMES and any form of EOMES resulting from processing in cells. The term also encompasses naturally occurring variants of EOMES, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human EOMES is set forth in SEQ ID NO: 3. The amino acid sequence of an exemplary protein encoded by human EOMES is set forth in SEQ ID NO: 4.

[0072] As used herein, the term "GZMA," unless otherwise indicated, refers to any native GZMA (granzyme A) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed GZMA and any form of GZMA that results from processing in cells. The term also encompasses naturally occurring variants of GZMA, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human GZMA is set forth in SEQ ID NO:51. The amino acid sequence of an exemplary protein encoded by human GZMA is set forth in SEQ ID NO:52.

[0073] As used herein, the term "GZMB," unless otherwise indicated, refers to any naturally occurring form of GZMB (Granzyme B) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed GZMB and any form of GZMB that results from processing in the cell. The term also encompasses naturally occurring variants of GZMB, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human GZMB is set forth in SEQ ID NO:53. The amino acid sequence of an exemplary protein encoded by human GZMB is set forth in SEQ ID NO:54.

[0074] As used herein, the term "PRF1," unless otherwise indicated, refers to any native PRF1 (also known as perforin 1, a pore-forming protein) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed PRF1 and any form of PRF1 resulting from processing in cells. The term also encompasses naturally occurring variants of PRF1, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human PRF1 is set forth in SEQ ID NO:5. The amino acid sequence of an exemplary protein encoded by human PRF1 is set forth in SEQ ID NO:6.

[0075] As used herein, the term "IFNG," unless otherwise indicated, refers to any naturally occurring IFNG (interferon, gamma) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed IFNG and any form of IFNG resulting from processing in cells. The term also encompasses naturally occurring variants of IFNG, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human IFNG is set forth in SEQ ID NO:7. The amino acid sequence of an exemplary protein encoded by human IFNG is set forth in SEQ ID NO:8.

[0076] The terms "programmed death-ligand 1" and "PD-L1," as used herein, refer to native sequence PD-L1 polypeptides, polypeptide variants, and fragments of native sequence polypeptides and polypeptide variants, as further defined herein. The PD-L1 polypeptides described herein may be isolated from a variety of sources, such as from human tissue types or from another source, or prepared by recombinant or synthetic methods.

[0077] A "native sequence PD-L1 polypeptide" includes a polypeptide having the same amino acid sequence as a corresponding PD-L1 polypeptide derived from nature. The term encompasses "full-length," unprocessed PD-L1 and any form of IFNG that results from processing in the cell. The term also encompasses naturally occurring variants of IFNG, such as splice variants or allelic variants.

[0078] The term "PD-L1 polypeptide variant" or variations thereof refers to a PD-L1 polypeptide, as defined herein, typically an active PD-L1 polypeptide, that has at least about 80% amino acid sequence identity to any of the native sequence PD-L1 polypeptide sequences disclosed herein. Such PD-L1 polypeptide variants include, for example, PD-L1 polypeptides with one or more amino acid residues added or deleted at the N- or C-terminus of the native amino acid sequence. Typically, a PD-L1 polypeptide variant will have at least about 80% amino acid sequence identity, or at least about 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity to a native sequence PD-L1 polypeptide sequence disclosed herein. Typically, PD-L1 variant polypeptides are at least about 10 amino acids in length, or at least about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 281, 282, 283, 284, 285, 286, 287, 288, or 289 amino acids in length or more. Optionally, the PD-L1 variant polypeptide has no more than one conservative amino acid substitution compared to a native PD-L1 polypeptide sequence, or no more than 2, 3, 4, 5, 6, 7, 8, 9, or 10 conservative amino acid substitutions compared to a native PD-L1 polypeptide sequence.

[0079] The term "vascular endothelial growth factor" or "VEGF" refers to vascular endothelial growth factor protein A, as exemplified by SwissProt accession number P15692, Gene ID (NCBI):7422. The term "VEGF" encompasses the protein having the amino acid sequence of SwissProt accession number P15692, Gene ID (NCBI):7422, as well as homologs and isoforms thereof. The term "VEGF" also refers to known isoforms of VEGF, e.g., splice isoforms, e.g., VEGF 111 , VEGF 121 , VEGF 145 , VEGF 165 , VEGF 189 , and VEGF 206 , as well as their naturally occurring alleles and processed forms (VEGF 165 These include VEGFs derived from non-human species such as mouse, rat, or primate. VEGFs derived from specific species may be designated by terms such as hVEGF for human VEGF and mVEGF for mouse VEGF. The term "VEGF" is also used to refer to truncated forms of the polypeptide, including amino acids 8-109 or 1-109 of the 165-amino acid human vascular endothelial growth factor. Reference to any such form of VEGF is referred to in this application as, for example, "VEGF." 109 ", "VEGF(8-109)", "VEGF(1-109)", or "VEGF 165". The amino acid positions of "truncated" native VEGF are numbered as shown in the native VEGF sequence. For example, amino acid position 17 (methionine) in truncated native VEGF is also position 17 (methionine) in native VEGF. Truncated native VEGF has binding affinity for KDR and Flt-1 receptors comparable to that of native VEGF. The term "VEGF variant," as used herein, refers to a VEGF polypeptide containing one or more amino acid mutations in the native VEGF sequence. In some cases, the one or more amino acid mutations include amino acid substitution(s). For purposes of concisely designating the VEGF variants described herein, it should be noted that the numbers refer to the amino acid residue position along the amino acid sequence of the putative native VEGF (provided in Leung et al., supra, and Houck et al., supra). Unless otherwise specified, the term "VEGF," as used herein, refers to VEGF-A.

[0080] The term "kinase insert domain receptor" or "KDR," as used herein, unless otherwise indicated, refers to any native KDR (also known in the art as fetal liver kinase 1 (FLK1) or vascular endothelial growth factor receptor 2 (VEGFR2)) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed KDR and any form of KDR resulting from processing in cells. The term also encompasses naturally occurring variants of KDR, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human KDR is set forth in SEQ ID NO:9. The amino acid sequence of an exemplary protein encoded by human KDR is set forth in SEQ ID NO:10.

[0081] The term "endothelial cell-specific molecule 1" or "ESM1," as used herein, unless otherwise indicated, refers to any native ESM1 (also known in the art as endocan) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed ESM1 and any form of ESM1 resulting from processing in cells. The term also encompasses naturally occurring variants of ESM1, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human ESM1 is set forth in SEQ ID NO: 11. The amino acid sequence of an exemplary protein encoded by human ESM1 is set forth in SEQ ID NO: 12.

[0082] As used herein, the term "platelet and endothelial cell adhesion molecule 1" or "PECAM1," unless otherwise indicated, refers to any native PECAM1 (also known in the art as CD31, endoCAM, GPIIA, or PECA1) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed PECAM1 and any form of PECAM1 that results from processing in cells. The term also encompasses naturally occurring variants of PECAM1, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human PECAM1 is set forth in SEQ ID NO: 13. The amino acid sequence of an exemplary protein encoded by human PECAM1 is set forth in SEQ ID NO: 14.

[0083] The term "FLT1," as used herein, unless otherwise indicated, refers to any native FLT1 (also known in the art as vascular endothelial growth factor receptor 1 (VEGFR1) or fms-related tyrosine kinase 1) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed FLT1 and any form of FLT1 that results from processing in cells. The term also encompasses naturally occurring variants of FLT1, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human FLT1 is set forth in SEQ ID NO:55. The amino acid sequence of an exemplary protein encoded by human FLT1 is set forth in SEQ ID NO:56.

[0084] The term "angiopoietin-like 4" or "ANGPTL4," as used herein, unless otherwise indicated, refers to any native ANGPTL4 (also known in the art as hepatic fibrinogen / angiopoietin-related protein (HFARP), peroxisome proliferator-activated receptor (PPAR) gamma, hepatic angiopoietin-related protein (HARP), angiopoietin-related protein 4 (Arp4), or fasting-induced adipose factor (FIAF)) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed ANGPTL4 and any form of ANGPTL4 resulting from processing in cells. The term also encompasses naturally occurring variants of ANGPTL4, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human ANGPTL4 is set forth in SEQ ID NO: 15. The amino acid sequence of an exemplary protein encoded by human ANGPTL4 is shown in SEQ ID NO:16.

[0085] The term "CD34," as used herein, unless otherwise indicated, refers to any native form of CD34 (also known in the art as the CD34 molecule or CD34 antigen) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CD34 and any form of CD34 that results from processing in cells. The term also encompasses naturally occurring variants of CD34, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CD34 is set forth in SEQ ID NO: 17. The amino acid sequence of an exemplary protein encoded by human CD34 is set forth in SEQ ID NO: 18.

[0086] As used herein, the term "interleukin-6" or "IL6," unless otherwise indicated, refers to any naturally occurring IL6 derived from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed IL6 and any form of IL6 resulting from processing in cells. The term also encompasses naturally occurring variants of IL6, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human IL6 is set forth in SEQ ID NO: 19. The amino acid sequence of an exemplary protein encoded by human IL6 is set forth in SEQ ID NO: 20.

[0087] The term "CXCL1," as used herein, unless otherwise indicated, refers to any native CXCL1 (also known as chemokine (C-X-C motif) ligand 1, GRO1, or neutrophil-activating protein 3 (NAP-3)) ​​from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCL1 and any form of CXCL1 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCL1, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCL1 is set forth in SEQ ID NO:21. The amino acid sequence of an exemplary protein encoded by human CXCL1 is set forth in SEQ ID NO:22.

[0088] As used herein, the term "CXCL2," unless otherwise indicated, refers to any native CXCL2 (also known as chemokine (C-X-C motif) ligand 2, macrophage inflammatory protein 2-alpha (MIP2-alpha)) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCL2 and any form of CXCL2 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCL2, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCL2 is set forth in SEQ ID NO: 23. The amino acid sequence of an exemplary protein encoded by human CXCL2 is set forth in SEQ ID NO: 24.

[0089] As used herein, the term "CXCL3," unless otherwise indicated, refers to any native CXCL3 (also known as chemokine (C-X-C motif) ligand 3, macrophage inflammatory protein 2-beta (MIP2-beta)) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCL3 and any form of CXCL3 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCL3, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCL3 is set forth in SEQ ID NO: 25. The amino acid sequence of an exemplary protein encoded by human CXCL3 is set forth in SEQ ID NO: 26.

[0090] The term "CXCL8," as used herein, unless otherwise indicated, refers to any native CXCL8 (also known as chemokine (C-X-C motif) ligand 8, interleukin 8 (IL8)) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCL8 and any form of CXCL8 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCL8, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCL8 is set forth in SEQ ID NO: 27. The amino acid sequence of an exemplary protein encoded by human CXCL8 is set forth in SEQ ID NO: 28.

[0091] As used herein, the term "PTGS2," unless otherwise indicated, refers to any native form of PTGS2 (also known as prostaglandin-endoperoxidase synthase 2, cyclooxygenase-2 (COX-2)) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed PTGS2 and any form of PTGS2 that results from processing in cells. The term also encompasses naturally occurring variants of PTGS2, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human PTGS2 is set forth in SEQ ID NO:29. The amino acid sequence of an exemplary protein encoded by human PTGS2 is set forth in SEQ ID NO:30.

[0092] As used herein, the term "CXCR1," unless otherwise indicated, refers to any native CXCR1 (also known as C-X-C motif chemokine receptor 1, interleukin-8 receptor, alpha, IL8RA, and CD181) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCR1 and any form of CXCR1 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCR1, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCR1 is set forth in SEQ ID NO: 75. The amino acid sequence of an exemplary protein encoded by human CXCR1 is set forth in SEQ ID NO: 76.

[0093] The term "CXCR2," as used herein, unless otherwise indicated, refers to any native CXCR2 (also known as C-X-C motif chemokine receptor 2, interleukin-8 receptor, beta, IL8RB, and CD182) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCR2 and any form of CXCR2 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCR2, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCR2 is set forth in SEQ ID NO: 77. The amino acid sequence of an exemplary protein encoded by human CXCR2 is set forth in SEQ ID NO: 78.

[0094] As used herein, the term "S100A8," unless otherwise indicated, refers to any native S100A8 (S100 calcium-binding protein A8, also known as calgranulin A) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). S100A8 can form a heterodimer with S100A9 called calprotectin. The term encompasses "full-length," unprocessed S100A8 and any form of S100A8 resulting from processing in cells. The term also encompasses naturally occurring variants of S100A8, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human S100A8 is set forth in SEQ ID NO:79. The amino acid sequence of an exemplary protein encoded by human S100A8 is set forth in SEQ ID NO:80.

[0095] The term "S100A9," as used herein, unless otherwise indicated, refers to any native S100A9 (also known as S100 calcium-binding protein A9, calgranulin B, and migration inhibitory-related protein 14 (MRP14)) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed S100A9 and any form of S100A9 resulting from processing in cells. The term also encompasses naturally occurring variants of S100A9, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human S100A9 is set forth in SEQ ID NO:81. The amino acid sequence of an exemplary protein encoded by human S100A9 is set forth in SEQ ID NO:82.

[0096] As used herein, the term "CXCL9," unless otherwise indicated, refers to any native CXCL9 (chemokine (C-X-C motif) ligand 9) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCL9 and any form of CXCL9 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCL9, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCL9 is set forth in SEQ ID NO:57. The amino acid sequence of an exemplary protein encoded by human CXCL9 is set forth in SEQ ID NO:58.

[0097] As used herein, the term "CXCL10," unless otherwise indicated, refers to any native CXCL10 (chemokine (C-X-C motif) ligand 10) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCL10 and any form of CXCL10 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCL10, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCL10 is set forth in SEQ ID NO:59. The amino acid sequence of an exemplary protein encoded by human CXCL10 is set forth in SEQ ID NO:60.

[0098] As used herein, the term "CXCL11," unless otherwise indicated, refers to any native CXCL11 (chemokine (C-X-C motif) ligand 11) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CXCL11 and any form of CXCL11 resulting from processing in cells. The term also encompasses naturally occurring variants of CXCL11, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CXCL11 is set forth in SEQ ID NO: 61. The amino acid sequence of an exemplary protein encoded by human CXCL11 is set forth in SEQ ID NO: 62.

[0099] The term "CD27," as used herein, unless otherwise indicated, refers to any native form of CD27 (also known in the art as CD27L receptor or TNFRSF7) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CD27 and any form of CD27 that results from processing in cells. The term also encompasses naturally occurring variants of CD27, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human CD27 is set forth in SEQ ID NO: 31. The amino acid sequence of an exemplary protein encoded by human CD27 is set forth in SEQ ID NO: 32.

[0100] As used herein, the term "FOXP3," unless otherwise indicated, refers to any native FOXP3 (forkhead box P3, also known in the art as scurfin) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed FOXP3 and any form of FOXP3 resulting from processing in cells. The term also encompasses naturally occurring variants of FOXP3, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human FOXP3 is set forth in SEQ ID NO: 33. The amino acid sequence of an exemplary protein encoded by human FOXP3 is set forth in SEQ ID NO: 34.

[0101] The term "PD-1," as used herein, unless otherwise indicated, refers to any native PD-1 (also known as PDCD1, programmed cell death protein 1, or CD279) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed PD-1 and any form of PD-1 that results from processing in cells. The term also encompasses naturally occurring variants of PD-1, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human PD-1 is set forth in SEQ ID NO:35. The amino acid sequence of an exemplary protein encoded by human PD-1 is set forth in SEQ ID NO:36.

[0102] The term "CTLA4," as used herein, unless otherwise indicated, refers to any native CTLA4 (cytotoxic T-lymphocyte-associated protein 4, also known in the art as CD152) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed CTLA4 and any form of CTLA4 that results from processing in cells. The term also encompasses naturally occurring variants of CTLA4, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human CTLA4 is set forth in SEQ ID NO:37. The amino acid sequence of an exemplary protein encoded by human CTLA4 is set forth in SEQ ID NO:38.

[0103] As used herein, the term "TIGIT," unless otherwise indicated, refers to any native TIGIT (a T cell immunoreceptor having Ig and ITIM domains) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed TIGIT and any form of TIGIT resulting from processing in cells. The term also encompasses naturally occurring variants of TIGIT, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human TIGIT is set forth in SEQ ID NO: 39. The amino acid sequence of an exemplary protein encoded by human TIGIT is set forth in SEQ ID NO: 40.

[0104] As used herein, the term "IDO1," unless otherwise indicated, refers to any native IDO1 (indoleamine 2,3-dioxygenase 1) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed IDO1 and any form of IDO1 resulting from processing in cells. The term also encompasses naturally occurring variants of IDO1, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human IDO1 is set forth in SEQ ID NO:41. The amino acid sequence of an exemplary protein encoded by human IDO1 is set forth in SEQ ID NO:42.

[0105] As used herein, the term "PSMB8" refers to any naturally occurring PSMB8 (proteasome subunit beta type 8) derived from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. The term encompasses "full-length," unprocessed PSMB8 and any form of PSMB8 resulting from processing in cells. The term also encompasses naturally occurring variants of PSMB8, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human PSMB8 is set forth in SEQ ID NO: 43. The amino acid sequence of an exemplary protein encoded by human PSMB8 is set forth in SEQ ID NO: 44.

[0106] As used herein, the term "PSMB9" refers to any naturally occurring PSMB9 (proteasome subunit beta 9) derived from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise indicated. The term encompasses "full-length," unprocessed PSMB9 and any form of PSMB9 resulting from processing in cells. The term also encompasses naturally occurring variants of PSMB9, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human PSMB9 is set forth in SEQ ID NO: 45. The amino acid sequence of an exemplary protein encoded by human PSMB9 is set forth in SEQ ID NO: 46.

[0107] The term "TAP1," as used herein, unless otherwise indicated, refers to any native form of TAP1 (transporter associated with antigen processing 1, also known in the art as antigenic peptide transporter 1) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed TAP1 and any form of TAP1 that results from processing in cells. The term also encompasses naturally occurring variants of TAP1, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human TAP1 is set forth in SEQ ID NO:47. The amino acid sequence of an exemplary protein encoded by human TAP1 is set forth in SEQ ID NO:48.

[0108] As used herein, the term "TAP2," unless otherwise indicated, refers to any native TAP2 (antigenic peptide transporter 2) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length," unprocessed TAP2 and any form of TAP2 resulting from processing in cells. The term also encompasses naturally occurring variants of TAP2, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human TAP2 is set forth in SEQ ID NO:49. The amino acid sequence of an exemplary protein encoded by human TAP2 is set forth in SEQ ID NO:50.

[0109] The terms "level of expression" or "expression level" are generally used interchangeably and generally refer to the amount of a biomarker in a biological sample. "Expression" generally refers to the process by which information (e.g., genetic code and / or epigenetic information) is converted into a structure present and functioning in a cell. Thus, as used herein, "expression" can refer to transcription into a polynucleotide, translation into a polypeptide, or even polynucleotide and / or polypeptide modification (e.g., post-translational modification of a polypeptide). Fragments of transcribed polynucleotides, translated polypeptides, or polynucleotide and / or polypeptide modifications (e.g., post-translational modification of a polypeptide) are also considered to be expressed, regardless of whether they are derived from transcripts generated by alternative splicing, degraded transcripts, or post-translational processing of a polypeptide, for example, by proteolysis. "Expressed genes" include those that are transcribed into polynucleotides as mRNA and then translated into polypeptides, and those that are transcribed into RNA but not translated into polypeptides (e.g., transfer and ribosomal RNA). The expression levels of two or more genes of interest can be determined by counting methods known to those skilled in the art and also disclosed herein, including, for example, calculating the median or mean value of all the expression levels of genes of interest.Before counting, the expression level of each gene of interest can be normalized by statistical methods known to those skilled in the art and also disclosed herein, including, for example, normalizing to the expression level of one or more housekeeping genes, or normalizing to the total library size, or normalizing to the median or mean value of the expression level values ​​of all genes measured.In some cases, before counting across multiple genes of interest, the normalized expression level of each gene of interest can be standardized by using statistical methods known to those skilled in the art and also disclosed herein (for example, by calculating the Z-score of the normalized expression level of each gene of interest).

[0110] A sample or cell that "expresses" a protein of interest is one in which the mRNA encoding the protein or the protein (including fragments thereof) is determined to be present in the sample or cell.

[0111] As used herein, the term "reference expression level" refers to an expression level in a sample from an individual that is compared to another expression level, e.g., the expression level of one or more genes described herein (e.g., any gene set forth in Table 1, or any combination thereof (e.g., any combination set forth in any one of Tables 2-12)), e.g., to make a predictive, diagnostic, prognostic, and / or therapeutic decision. For example, a reference expression level may be a level that is determined based on expression levels in a reference population (e.g., the median expression level in a reference population, e.g., a population of patients with cancer), a reference sample, and / or a pre-specified value (e.g., a significant difference (above a cutoff value and / or below a cutoff value) between the individual's responsiveness to treatment with an anti-cancer therapy and the individual's responsiveness to treatment with a different anti-cancer therapy) in a reference population. For example, the reference population may be derived from a predetermined cutoff value that significantly (e.g., statistically significantly) separates a first subset of individuals treated with an anti-cancer therapy comprising a PD-L1-binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab (MPDL3280A)), or a PD-1-binding antagonist (e.g., an anti-PD-1 antibody), or an anti-cancer therapy comprising a multi-targeted tyrosine kinase inhibitor) from a second subset of individuals in the same reference population who are treated with a different anti-cancer therapy (or who have not been treated with an anti-cancer therapy). In some embodiments, the cutoff value may be the median or mean expression level in the reference population. In other embodiments, the reference level may be in the top 40%, top 30%, top 20%, top 10%, top 5%, or top 1% of expression levels in the reference population. In certain embodiments, the cutoff value may be the median expression level in the reference population.One of skill in the art will understand that the numerical values ​​of the reference expression levels may vary depending on the indication (e.g., cancer (e.g., kidney cancer, breast cancer, lung cancer, or bladder cancer), the methodology used to detect expression levels (e.g., RNA-seq or RT-qPCR), and / or the particular combination of genes tested (e.g., any combination of genes listed in Table 1, or any one of the combinations of genes listed in Tables 2-12).

[0112] The terms "above" a level (e.g., above a reference level), "increased expression," "increased expression level," "increased level," "elevated expression," "elevated expression level," or "elevated level" refer to the expression level of a biomarker compared to the expression level of a biomarker in a control (e.g., individual(s) not afflicted with a disease or disorder (e.g., cancer)), an internal control (e.g., housekeeping biomarker), or the level of a biomarker in a sample obtained before administration of a treatment (e.g., an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 antagonist), or compared to a reference level. It refers to increased expression or increased level of a biomarker in an individual compared to (e.g., the median expression level of the biomarker in samples from a group / population of patients (e.g., patients with cancer who are being tested for responsiveness to a VEGF antagonist and a PD-L1 axis-binding antagonist), the median expression level of the biomarker in samples from a group / population of patients (e.g., patients with cancer who are identified as not responsive to a VEGF antagonist and a PD-L1 axis-binding antagonist), or the level in a sample previously obtained from the individual).

[0113] The terms "below" (e.g., below a reference level), "decreased expression," "decreased expression level," "reduced level," "reduced expression," "reduced expression level," or "reduced level" refer to the expression level of a biomarker compared to a control (e.g., an individual(s) not afflicted with a disease or disorder (e.g., cancer)), an internal control (e.g., a housekeeping biomarker), or the level of a biomarker in a sample obtained before administration of a treatment (e.g., an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 antagonist), or compared to a reference level. "Decreased expression" refers to decreased expression or a decreased level of a biomarker in an individual compared to (e.g., the median expression level of the biomarker in samples from a group / population of patients (e.g., patients with cancer who are being tested for responsiveness to a VEGF antagonist and a PD-L1 axis-binding antagonist), the median expression level of the biomarker in samples from a group / population of patients (e.g., patients with cancer who are identified as not responsive to a VEGF antagonist and a PD-L1 axis-binding antagonist), or the level in a sample previously obtained from the individual. In some embodiments, reduced expression is little or no expression.

[0114] As used herein, a "reference sample," "reference cell," "reference tissue," "control sample," "control cell," or "control tissue" refers to a sample, cell, tissue, or standard 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-diseased part of the body (e.g., tissue or cells) of the same patient or individual. For example, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue may be a healthy and / or non-diseased cell or tissue adjacent to the diseased cell or tissue (e.g., a cell or tissue adjacent to a tumor). In another embodiment, the reference sample is obtained from an untreated tissue and / or cell of the body of the same patient or individual. 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-diseased part of the body (e.g., tissue or cells) of an individual other than the patient or individual. In yet another embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from untreated tissues and / or cells from the body of an individual that is not the patient or individual. In another embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from the patient prior to administration of a treatment (e.g., an anti-cancer treatment comprising a VEGF antagonist and / or a PD-L1 axis-binding antagonist).

[0115] The phrase "based on," as used herein, means that information about one or more biomarkers is used to inform treatment decisions, information provided in package inserts, marketing / promotional guidelines, etc.

[0116] The term "housekeeping biomarker" refers to a biomarker or group of biomarkers (e.g., polynucleotides and / or polypeptides) that are typically present in the same way in all cell types. In some embodiments, a housekeeping biomarker is a "housekeeping gene." "Housekeeping gene," as used herein, refers to a gene or group of genes that encode proteins whose activity is essential for maintaining cellular function and that are typically present in the same way in all cell types.

[0117] "Correlate" or "correlating" means comparing, in any manner, the performance and / or results of a first analysis or protocol with the performance and / or results of a second analysis or protocol. For example, the results of a first analysis or protocol can be used in performing a second protocol and / or can be used to determine whether a second analysis or protocol should be performed. With respect to polypeptide analysis or protocol embodiments, the results of a polypeptide expression analysis or protocol can be used to determine whether a particular therapeutic regimen should be performed. With respect to polynucleotide analysis or protocol embodiments, the results of a polynucleotide expression analysis or protocol can be used to determine whether a particular therapeutic regimen should be performed.

[0118] As used herein, "treatment" (and grammatical variations thereof, such as "treat" or "treating") refers to a clinical intervention aimed at altering the natural course of the individual being treated and may be performed prophylactically or during the course of clinical pathology. Desired effects of treatment include prevention of disease onset or recurrence, alleviation of symptoms, attenuation of any direct or indirect pathological consequences of the disease, prevention of metastasis, slowing the rate of disease progression, remission or palliation of disease symptoms, and recovery or improved prognosis. In some embodiments, antibodies (e.g., anti-VEGF antibodies, and anti-PD-L1 antibodies, or anti-PD-1 antibodies) are used to delay disease development or slow the progression of a disease or disorder.

[0119] As used herein, "amplification" generally refers to the process of generating multiple copies of a desired sequence. "Multiple copies" means at least two copies. "Copy" does not necessarily mean perfect sequence complementarity or identity to the template sequence. For example, copies may contain nucleotide analogs such as deoxyinosine, intentional sequence modifications (such as those introduced by primers containing sequences that are hybridizable to but not complementary to the template), and / or sequence errors that occur during amplification.

[0120] The term "multiplex PCR" refers to a single PCR reaction performed on nucleic acids obtained from a single source (e.g., an individual) using two or more primer sets to amplify two or more DNA sequences in a single reaction.

[0121] As used herein, the term "polymerase chain reaction" or "PCR" generally refers to a procedure in which minute, specific pieces of nucleic acid, RNA, and / or DNA are amplified, as described, for example, in U.S. Pat. No. 4,683,195. Generally, sequence information at or beyond the ends of the region of interest must be available so that oligonucleotide primers can be designed; the sequences of these primers are identical or similar to those of opposite strands of the template to be amplified. The 5' terminal nucleotides of the two primers may correspond to the ends of the material to be amplified. PCR can be used to amplify specific RNA sequences, specific DNA sequences from total genomic DNA, cDNA transcribed from total cellular RNA, bacteriophage sequences, or plasmid sequences, etc. See generally Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263 (1987) and Erlich, ed., PCR Technology (Stockton Press, NY, 1989). As used herein, PCR is considered to be one example, but not the only example, of a nucleic acid polymerase reaction method for amplifying a nucleic acid test sample, which involves the use of known nucleic acids (DNA or RNA) as primers, and utilizes a nucleic acid polymerase to amplify or generate a specific piece of nucleic acid, or a specific piece of nucleic acid complementary to a specific nucleic acid.

[0122] "Quantitative real-time polymerase chain reaction" or "qRT-PCR" refers to a form of PCR in which the amount of PCR product is measured at each step of the PCR reaction. This technique has been described in various publications, including, for example, Cronin et al., Am. J. Pathol. 164(1):35-42 (2004) and Ma et al., Cancer Cell 5:607-616 (2004).

[0123] The term "microarray" refers to an ordered arrangement of hybridizable array elements, preferably polynucleotide probes, on a substrate.

[0124] The term "RNA-seq," also known as "whole transcriptome shotgun sequencing (WTSS)," refers to the use of high-throughput sequencing techniques to sequence and / or quantify cDNA to obtain information about the RNA content of a sample. Publications describing RNA-seq include Wang et al., Nature Reviews Genetics 10(1):57-63, 2009; Ryan et al., BioTechniques 45(1):81-94, 2008; and Maher et al., Nature 458(7234):97-101, 2009.

[0125] The term "diagnosis" is used herein to refer to the identification or classification of a molecular or pathological state, disease, or condition, such as cancer (e.g., kidney cancer). For example, "diagnosis" can refer to the identification of a particular type of cancer. "Diagnosis" can also refer to the classification of a particular subtype of cancer (e.g., a subtype characterized by the expression of one or a combination of biomarkers (e.g., particular genes, or proteins encoded by said genes)), for example, by histopathological criteria or by molecular features. In some embodiments, the diagnosis is of a sarcomatoid cancer (e.g., sarcomatoid kidney cancer (e.g., sarcomatoid RCC)).

[0126] "Tumor-infiltrating immune cells," as used herein, refer to any immune cells present in a tumor or a sample thereof. Tumor-infiltrating immune cells include, but are not limited to, intratumoral immune cells, peritumoral immune cells, other tumor stromal cells (e.g., fibroblasts), or any combination thereof. Such tumor-infiltrating immune cells include, for example, T lymphocytes (e.g., CD8 + T lymphocytes and / or CD4 + T lymphocytes), B lymphocytes, or other myeloid lineage cells, such as granulocytes (e.g., neutrophils, eosinophils, and basophils), monocytes, macrophages (e.g., CD68 + / CD163 +macrophages), dendritic cells (e.g., interdigitating dendritic cells), histiocytes, and natural killer (NK) cells.

[0127] As used herein, "tumor cell" refers to any tumor cell present in a tumor or a sample thereof. Tumor cells can be distinguished from other cells that may be present in a tumor sample, such as stromal cells and tumor-infiltrating immune cells, using methods known in the art and / or described herein.

[0128] As used herein, "administering" refers to administering a dosage of a compound (e.g., a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab) or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), a PD-L1 axis-binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab), and / or an angiogenesis inhibitor (e.g., a VEGF antagonist (e.g., a VEGFR inhibitor (e.g., or a composition (e.g., a pharmaceutical composition, e.g., a pharmaceutical composition comprising a VEGF antagonist, a PD-L1 axis binding antagonist, and / or an angiogenesis inhibitor (e.g., a VEGF antagonist (e.g., a VEGFR inhibitor, (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))))) to a patient. The compositions used in the methods described herein can be administered, for example, intramuscularly, intravenously, intradermally, transdermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intrathecally, intranasally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctivally, intravesicularly, mucosally, intrapericardially, intraumbilically, intraocularly, intraorbitally, intravitreally (for example, by intravitreal injection), by eye drops, orally, topically, transdermally, parenterally, by inhalation, by injection, by implantation, by infusion, by continuous infusion, by local perfusion that directly bathes target cells, by catheter, by lavage, in cream, or in lipid compositions.The compositions used in the methods described herein can also be administered systemically or locally.The administration method can vary depending on various factors (for example, the compound or composition to be administered and the severity of the condition, disease, or disorder to be treated).

[0129] A "therapeutically effective amount" refers to the amount of a therapeutic agent for treating or preventing a disease or disorder (e.g., cancer, e.g., renal cancer (e.g., RCC)) in a mammal (e.g., a human). In the case of cancer, a therapeutically effective amount of a therapeutic agent can reduce the number of cancer cells, reduce primary tumor size, inhibit (i.e., slow down to some extent, and preferably stop) cancer cell invasion into peripheral organs, inhibit (i.e., slow down to some extent, and preferably stop) tumor metastasis, inhibit tumor growth to some extent, and / or alleviate to some extent one or more symptoms associated with the disorder. To the extent that a drug can prevent the growth of and / or kill existing cancer cells, the drug can be cytostatic and / or cytotoxic. With respect to cancer therapy, in vivo efficacy may be measured, for example, by assessing duration of survival (e.g., overall survival or progression-free survival), time to disease progression (TTP), response rate (e.g., overall response (ORR), complete response (CR), and partial response (PR)), duration of response, progression-free rate (DFR), and / or quality of life.

[0130] The term "concurrently" is used herein to refer to the administration of two or more therapeutic agents, with at least partial overlap in administration time. Thus, concurrent administration includes a dosing regimen in which the administration of one or more agents continues after the administration of one or more other agents is discontinued. For example, in some embodiments, a VEGF antagonist and a PD-L1 axis-binding antagonist can be administered simultaneously.

[0131] "Reduce or inhibit" refers to the ability to cause an overall decrease of 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or more. Reduction or inhibition may refer, for example, to symptoms of the disorder being treated, the presence or size of metastases, or the size of the primary tumor.

[0132] As used herein, a "loading" dose generally refers to the initial dose of a therapeutic agent administered to a patient, followed by one or more maintenance doses thereof. Generally, a single loading dose is administered, although multiple loading doses are contemplated herein. Typically, the amount of the administered loading dose(s) exceeds the amount of the administered maintenance dose(s), and / or the loading dose(s) are administered more frequently than the maintenance dose(s) so as to achieve the desired steady-state concentration of the therapeutic agent more quickly than can be achieved with the maintenance dose(s).

[0133] A "maintenance" dose or an "extended" dose herein refers to one or more doses of a therapeutic agent administered to a patient over a treatment period. Typically, maintenance doses are administered at spaced treatment intervals, such as about every week, about every two weeks, about every three weeks, or about every four weeks.

[0134] "Response to treatment," "responsiveness to treatment," or "benefit from treatment" can be assessed using any endpoint that indicates benefit to an individual, including, but not limited to: (1) inhibition to some extent of disease progression (e.g., cancer progression), including slowing and complete halt; (2) reduction in tumor size; (3) inhibition (i.e., reduction, slowing, or complete halt) of cancer cell invasion into adjacent peripheral organs and / or tissues; (4) inhibition (i.e., reduction, slowing, or complete halt) of metastasis; (5) alleviation to some extent of one or more symptoms associated with a disease or disorder (e.g., cancer); (6) overall survival (OS HR<1), progression-free survival (PFS), or (7) an increase in overall response rate (ORR), complete response (CR) rate, and / or progression-free rate (DFR); and / or (8) a significant improvement in survival after treatment (e.g., a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab) or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., A reduction in mortality at a given time point following treatment with an anti-cancer therapy comprising a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab (MPDL3280A)) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), or an angiogenesis inhibitor (e.g., a VEGF antagonist (e.g., a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))).

[0135] "Response" refers to a measurable response, including complete response (CR) or partial response (PR). In some embodiments, "objective response rate (ORR)" refers to the sum of the complete response (CR) rate and the partial response (PR) rate.

[0136] By "complete response" or "CR" is intended the disappearance of all signs of cancer (e.g., disappearance of all target lesions) in response to treatment. This does not necessarily mean that the cancer has been cured.

[0137] As used herein, "partial response" or "PR" refers to a decrease in the size of one or more tumors or lesions, or the extent of cancer within the body, in response to treatment. For example, in some embodiments, PR refers to at least a 30% decrease in the sum of the longest diameters (SLD) of target lesions, relative to the baseline SLD.

[0138] "Sustained response" refers to a sustained effect on reducing tumor growth after treatment has ceased. For example, the tumor size may remain the same or become smaller compared to the size at the beginning of the administration period. In some embodiments, the sustained response has a duration at least equal to the treatment period, or at least 1.5, 2.0, 2.5, or 3.0 times the treatment period or more.

[0139] As used herein, "stable disease" or "SD" refers to neither sufficient shrinkage of target lesions to qualify as PR nor sufficient increase to qualify as PD, based on the smallest SLD since treatment initiation.

[0140] As used herein, "progressive disease" or "PD" refers to at least a 20% increase in the SLD of a target lesion relative to the smallest SLD recorded since the start of treatment or the presence of one or more new lesions.

[0141] The term "survival" refers to the patient being alive, and includes overall survival and progression-free survival.

[0142] As used herein, "progression-free survival" (PFS) refers to the length of time during and after treatment during which the disease being treated (e.g., cancer, e.g., renal cancer (e.g., RCC)) does not worsen or worsen. Progression-free survival can include the amount of time an individual experiences a complete or partial response, and the amount of time an individual experiences stable disease.

[0143] As used herein, "overall survival" or "OS" refers to the percentage of subjects in a population who are likely alive after a particular period of time (e.g., 6 months, 1 year, 2 years, 3 years, 4 years, 5 years, 10 years, 15 years, 20 years, or more than 20 years from the time of diagnosis or treatment).

[0144] "Prolonged survival" means an increase in overall survival or progression-free survival in a treated patient compared to untreated patients (i.e., compared to patients not treated with the pharmaceutical agent), or compared to patients who do not express the biomarker at a specified level, and / or compared to patients who are treated with an approved anti-tumor agent (e.g., an anti-VEGF antibody (e.g., bevacizumab), a PD-L1 axis-binding antagonist (e.g., atezolizumab), and / or a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib)).

[0145] As used herein, "hazard ratio" or "HR" is a statistical definition of the occurrence rate of an event. For the purpose of the present invention, hazard ratio is defined as the probability of an event (e.g., PFS or OS) in an experimental (e.g., treatment) group / arm divided by the probability of the event in a control group / arm at any particular time point. An HR value of 1 indicates that the relative risk of an endpoint (e.g., death) is equal in both the "treatment" group and the "control" group, a value greater than 1 indicates that the risk is greater in the treatment group compared to the control group, and a value less than 1 indicates that the risk is greater in the control group compared to the treatment group. In progression-free survival analysis, the "hazard ratio" (i.e., PFS HR) is a summary of the difference between two progression-free survival curves, representing the reduction in the risk of death in treatment compared to control over the follow-up period. In overall survival analysis, the "hazard ratio" (i.e., OS HR) is a summary of the difference between two overall survival curves, representing the reduction in the risk of death in treatment compared to control over the follow-up period.

[0146] As used herein, "deterioration-free rate" or "DFR" refers to the probability that a patient will experience a clinically meaningful deterioration over a given length of time (e.g., the time from the start of treatment to the patient's first increase of 2 or more points above baseline on the MD Anderson Symptom Assessment Inventory (MDASI) Interference scale).

[0147] The "MD Anderson Symptom Assessment Inventory (MDASI) Interference Scale" refers to a patient-reported outcome measurement scoring system that assesses the severity and impact of multiple symptoms related to cancer and its treatment (see, e.g., Mendoza et al., Clin. Breast Cancer 13:325–334, 2013; Jones et al., Clin. Genitourin. Cancer 12:41–49, 2014; and Shi et al., Pain 158:1108–1112, 2017). On the MDASI Interference Scale, patients rate how much their symptoms have interfered with various aspects of their life over the past 24 hours. Each interference item (work, general activities, walking, relationships with others, enjoyment of life, and mood) is rated on a scale of 0–10, with 0 representing "not interfering" and 10 representing "completely interfering."

[0148] The term "anti-cancer therapy" refers to a therapy useful in the treatment of cancer. Examples of anti-cancer therapeutic agents include cytotoxic agents, chemotherapeutic agents, growth inhibitory agents, agents used in radiation therapy, anti-angiogenic agents, apoptotic agents, anti-tubulin agents, and other agents for treating cancer, such as, but not limited to, anti-CD20 antibodies, platelet-derived growth factor inhibitors (e.g., GLEEVEC™ (imatinib mesylate)), COX-2 inhibitors (e.g., celecoxib), interferons, cytokines, antagonists (e.g., neutralizing antibodies) that bind to one or more of the following targets: PDGFR-β, BlγS, APRIL, BCMA receptor(s), TRAIL / Apo2, other bioactive and organic chemical agents, and the like. Combinations of these are also encompassed by the present invention.

[0149] The term "VEGF antagonist" or "VEGF-specific antagonist" refers to a molecule capable of binding to VEGF, reducing VEGF expression levels, or neutralizing, blocking, inhibiting, abrogating, reducing, or interfering with the biological activities of VEGF (including, but not limited to, VEGF binding to one or more VEGF receptors, VEGF signal transduction, and VEGF-mediated angiogenesis, and endothelial cell survival or proliferation). For example, a molecule capable of neutralizing, blocking, inhibiting, abrogating, reducing, or interfering with the biological activities of VEGF may exert its effect by binding to one or more VEGF receptors (VEGFRs) (e.g., VEGFR1, VEGFR2, VEGFR3, membrane-bound VEGF receptors (mbVEGFRs), or soluble VEGF receptors (sVEGFRs)). Such antagonists are also referred to herein as "VEGFR inhibitors." VEGF-specific antagonists useful in the methods of the present invention include polypeptides that specifically bind to VEGF, anti-VEGF antibodies and antigen-binding fragments thereof, receptor molecules and derivatives that specifically bind to VEGF and thereby block binding to one or more receptors, fusion proteins (e.g., VEGF-Trap (Regeneron)), and VEGF receptor molecules. 121-Gelonin (Peregrine). VEGF-specific antagonists also include antagonist variants of VEGF polypeptides, antisense nucleobase oligomers complementary to at least a fragment of a nucleic acid molecule encoding a VEGF polypeptide, small RNAs complementary to at least a fragment of a nucleic acid molecule encoding a VEGF polypeptide, ribozymes targeting VEGF, peptibodies against VEGF, and VEGF aptamers. VEGF antagonists also include polypeptides that bind to VEGFR, anti-VEGFR antibodies and antigen-binding fragments thereof, as well as derivatives or fusion proteins that bind to VEGFR and thereby block, inhibit, abrogate, reduce, or interfere with VEGF biological activity (e.g., VEGF signal transduction). VEGF-specific antagonists also include non-peptide small molecules that can bind to VEGF or VEGFR and block, inhibit, abrogate, reduce, or interfere with VEGF biological activity. Thus, the term "VEGF activity" specifically includes VEGF-mediated biological activity of VEGF. In certain embodiments, the VEGF antagonist reduces or inhibits the expression level or biological activity of VEGF by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. In some embodiments, the VEGF inhibited by the VEGF-specific antagonist is VEGF(8-109), VEGF(1-109), or VEGF 165 is.

[0150] As used herein, VEGF antagonists include anti-VEGFR2 antibodies and related molecules (e.g., ramucirumab, tanibirumab, aflibercept), anti-VEGFR1 antibodies and related molecules (e.g., icrucumab, aflibercept (VEGF Trap-Eye, EYLEA®), and dib-aflibercept (VEGF Trap, ZALTRAP®), bispecific VEGF antibodies (e.g., MP-0250, vanucizumab (VEGF-ANG2), and the bispecific antibodies disclosed in US2001 / 0236388), bispecific antibodies comprising a combination of two of an anti-VEGF arm, an anti-VEGFR1 arm, and an anti-VEGFR2 arm, anti-VEGFA antibodies (e.g., bevacizumab, sevacizumab), anti-VEGFB antibodies, anti-VEGF antibodies, and the like. These include, but are not limited to, GFC antibodies (e.g., VGX-100), anti-VEGFD antibodies, and non-peptide small molecule VEGF antagonists (e.g., pazopanib, axitinib, vandetanib, stivarga, cabozantinib, lenvatinib, nintedanib, orantinib, telatinib, dovitinib, cediranib, motesanib, surufatinib, apatinib, foretinib, famitinib, and tivozanib).

[0151] An "anti-VEGF antibody" is an antibody that binds to VEGF with sufficient affinity and specificity. In certain embodiments, the antibody will have a sufficiently high binding affinity for VEGF, e.g., the antibody may bind to hVEGF with a Kd value of 100 nM to 1 pM. Antibody affinity can be determined, for example, by surface plasmon resonance-based assays (such as BIAcore® assays as described in PCT Application Publication No. WO 2005 / 012359), enzyme-linked immunosorbent assays (ELISAs), and competitive assays (e.g., radioimmunoassays (RIAs)).

[0152] In certain embodiments, anti-VEGF antibodies can be used as therapeutic agents to target and interfere with diseases or conditions involving VEGF activity. Antibodies can also be subjected to other biological activity assays, for example, to evaluate their effectiveness as therapeutic agents. Such assays are known in the art and depend on the target antigen and intended use of the antibody. Examples include HUVEC inhibition assays, tumor cell growth inhibition assays (e.g., those described in WO 89 / 06692), antibody-dependent cellular cytotoxicity (ADCC) and complement-mediated cytotoxicity (CDC) assays (U.S. Pat. No. 5,500,362), and agonist activity or hematopoietic assays (see WO 95 / 27062). Anti-VEGF antibodies typically do not bind to other VEGF homologs, such as VEGF-B or VEGF-C, or other growth factors, such as PIGF, PDGF, or bFGF. In one embodiment, the anti-VEGF antibody is a monoclonal antibody that binds to the same epitope as monoclonal anti-VEGF antibody A4.6.1, produced by hybridoma ATCC HB 10709. In another embodiment, the anti-VEGF antibody is a recombinant humanized anti-VEGF monoclonal antibody produced according to Presta et al. (Cancer Res. 57:4593-4599, 1997), including, but not limited to, the antibody known as bevacizumab (BV, AVASTIN®).

[0153] The anti-VEGF antibody "bevacizumab (BV)," also known as "rhuMAb VEGF" or "AVASTIN®," is a recombinant humanized anti-VEGF monoclonal antibody produced according to Presta et al. (Cancer Res. 57:4593-4599, 1997). It contains mutated human IgG1 framework regions and antigen-binding complementarity-determining regions derived from the murine anti-hVEGF monoclonal antibody A.4.6.1, which blocks binding of human VEGF to its receptor. Approximately 93% of the amino acid sequence of bevacizumab, including most of the framework regions, is derived from human IgG1, and approximately 7% of the sequence is derived from the murine antibody A4.6.1. Bevacizumab has a molecular weight of approximately 149,000 daltons and is glycosylated. Bevacizumab and other humanized anti-VEGF antibodies are further described in U.S. Patent No. 6,884,879, issued February 26, 2005, the entire disclosure of which is expressly incorporated herein by reference. Additional preferred antibodies include the G6 or B20 series antibodies (e.g., G6-31, B20-4.1), described in PCT Application Publication No. WO2005 / 012359. For additional preferred antibodies, see U.S. Patent Nos. 7,060,269, 6,582,959, 6,703,020, 6,054,297, WO98 / 45332, WO96 / 30046, WO94 / 10202, EP0666868B1, U.S. Patent Application Publication Nos. 2006009360, 20050186208, 20030206899, 20030190317, 20030203409, and 20050112126, and Popkov et al. (Journal of Immunological Methods 288:149-164, 2004). Other preferred antibodies include antibodies that bind to a functional epitope on human VEGF that includes residues F17, M18, D19, Y21, Y25, Q89, 191, K101, E103, and C104, or alternatively includes residues F17, Y21, Q22, Y25, D63, 183, and Q89.

[0154] The term "PD-L1 axis-binding antagonist" refers to a molecule that inhibits the interaction of a PD-L1 axis-binding partner with one or more of its binding partners, so as to eliminate T cell dysfunction resulting from signaling on the PD-1 signaling axis, resulting in restored or enhanced T cell function. As used herein, PD-L1 axis-binding antagonists include PD-L1 binding antagonists and PD-1 binding antagonists, as well as molecules that interfere with the interaction between PD-L1 and PD-1 (e.g., PD-L2-Fc fusions).

[0155] The terms "anti-PD-L1 antibody" and "antibody that binds to PD-L1" refer to an antibody that can bind to PD-L1 with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent in targeting PD-L1. In one embodiment, the extent of binding of the anti-PD-L1 antibody to an unrelated, non-PD-L1 protein is less than about 10% of the antibody's binding to PD-L1, as measured, for example, by RIA. In certain embodiments, the anti-PD-L1 antibody binds to an epitope of PD-L1 that is conserved among PD-L1 from different species.

[0156] The terms "anti-PD-1 antibody" and "antibody that binds to PD-1" refer to an antibody that can bind to PD-1 with sufficient affinity so that the antibody is useful as a diagnostic and / or therapeutic agent in targeting PD-1. In one embodiment, the extent of binding of the anti-PD-1 antibody to an unrelated, non-PD-1 protein is less than about 10% of the antibody's binding to PD-1, as measured, for example, by RIA. In certain embodiments, the anti-PD-1 antibody binds to an epitope of PD-1 that is conserved among PD-1 from different species.

[0157] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, abrogates, or prevents signaling resulting from the interaction of PD-L1 with any one or more of its binding partners (e.g., PD-1 or B7-1). In some embodiments, a PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partners. In particular aspects, PD-L1 binding antagonists inhibit the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, PD-L1 binding antagonists include anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, abrogate, or prevent signaling resulting from the interaction of PD-L1 with one or more of its binding partners (e.g., PD-1 or B7-1). In one embodiment, the PD-L1 binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes that mediate PD-L1-mediated signaling, alleviating the dysfunctional state of dysfunctional T cells (e.g., enhancing 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 (CAS Registry Number: 1422185-06-5), also known as MPDL3280A and described herein. In another specific embodiment, the anti-PD-L1 antibody is YW243.55.S70, described herein. In another specific embodiment, the anti-PD-L1 antibody is MDX-1105, described herein. In yet another specific aspect, the anti-PD-L1 antibody is MEDI4736 (durvalumab), described herein. In yet another specific embodiment, the anti-PD-L1 antibody is MSB0010718C (avelumab) described herein.

[0158] As used herein, a "PD-1 binding antagonist" is a molecule that reduces, blocks, inhibits, abrogates, or interferes with signal transduction resulting from the interaction of PD-1 with one or more of its binding partners (e.g., PD-L1 and / or PD-L2). In some embodiments, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to its binding partners. In particular aspects, a PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and / or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies and antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, small molecule antagonists, polynucleotide antagonists, and other molecules that reduce, block, inhibit, abrogate, or interfere with signal transduction resulting from the interaction of PD-1 with PD-L1 and / or PD-L2. In one embodiment, the PD-1 binding antagonist reduces negative signals mediated by or through cell surface proteins expressed on T lymphocytes and other cells that mediate signaling through PD-1 or PD-L1, thereby alleviating the dysfunctional state of dysfunctional T cells. In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In a specific aspect, the PD-1 binding antagonist is MDX-1106 (nivolumab). In another specific aspect, the PD-1 binding antagonist is MK-3475 (pembrolizumab). In another specific aspect, the PD-1 binding antagonist is MEDI-0680 (AMP-514). In another specific aspect, the PD-1 binding antagonist is PDR001. In another specific aspect, the PD-1 binding antagonist is REGN2810. In another specific aspect, the PD-1 binding antagonist is BGB-108. In another specific embodiment, the PD-1 binding antagonist is AMP-224.

[0159] "Angiogenesis inhibitor" or "anti-angiogenic agent" refers to a low molecular weight substance, polynucleotide, polypeptide, isolated protein, recombinant protein, antibody, or conjugate or fusion protein thereof that directly or indirectly inhibits angiogenesis, vasculogenesis, or unwanted vascular permeability. It should be understood that anti-angiogenic agents include agents that bind to and block the angiogenic activity of an angiogenic factor or its receptor. For example, anti-angiogenic agents are antibodies or other antagonists against the angiogenic agents defined above, such as antibodies against VEGF-A or VEGF-A receptors (e.g., KDR receptor or Flt-1 receptor), anti-PDGFR inhibitors such as GLEEVEC™ (imatinib mesylate). Anti-angiogenic agents also include natural angiogenesis inhibitors, such as angiostatin, endostatin, etc. See, e.g., Klagsbrun and D'Amore, Annu. Rev. Physiol., 53:217-39 (1991); Streit and Detmar, Oncogene, 22:3172-3179 (2003) (e.g., Table 3 listing antiangiogenic treatments in malignant melanoma); Ferrara & Alitalo, Nature Medicine 5(12):1359-1364 (1999); Tonini et al., Oncogene, 22:6549-6556 (2003); and Sato, Int. J. Clin. Oncol., 8:200-206 (2003).

[0160] The term "cytotoxic agent," as used herein, refers to a substance that inhibits or prevents the function of cells and / or causes destruction of cells. This term includes radioactive isotopes (e.g., At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212 , P 32, and radioactive isotopes of Lu), chemotherapeutic agents such as methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin, or other intercalating agents, enzymes such as nucleases and fragments thereof, antibiotics, toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant, or animal origin (including fragments and / or variants thereof), and various antitumor or anticancer agents disclosed below. Tumoricidal agents cause the destruction of tumor cells.

[0161] A "chemotherapeutic agent" includes chemical compounds useful in the treatment of cancer. Examples of chemotherapeutic agents include erlotinib (TARCEVA®, Genentech / OSI Pharm.), bortezomib (VELCADE®, Millennium), and erythropoietinib (ERT). 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), finasunate (VATALANIB®, Novartis), oxaliplatin (ELOXATIN®, Sanofi), 5-FU (5-fluorouracil), leucovorin, rapamycin (sirolimus, RAPAMUNE®, Pfizer), lapatinib (TYKERB®, GSK572016, GlaxoSmithKline), Smith Kline), lonafamib (SCH66336), sorafenib (NEXAVAR®, Bayer Labs), gefitinib (IRESSA®, AstraZeneca), AG1478, alkylating agents (such as thiotepa and CYTOXAN® cyclophosphamide), alkylsulfonates (such as busulfan, improsulfan, and piposulfan), aziridines (such as benzodopa, carboquone, methyledopa, and uredopa), ethyleneimines and methylmelamines (altretamine, triethylene melamine, triethylene phosphoramide, triethylenethiophosphoramide, and trimethylmelamine), acetogenins (especially bullatacin and bullatacinone), camptothecins (including topotecan and irinotecan), bryostatin, kallistatin, CC-1065 (including its azozelesin, carzelesin, and bizelesin synthetic analogs), cryptophycins (especially cryptophycin 1 and cryptophycin 8), corticosteroids (including prednisone and prednisolone), cyproterone acetate,5α-reductase inhibitors (including finasteride and dutasteride), vorinostat, romidepsin, panobinostat, valproic acid, mocetinostat, dolastatins, aldesleukin, talc duocarmycins (including synthetic analogs KW-2189 and CB1-TM1), eluterobin, pancratistatin, sarcodictyin, spongistatin, nitrogen mustards (chlorambucil, chromafazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melfala antibiotics such as benzodiazepines (e.g., benzodiazepines, benzocaine, benzodiazepines, benzocaine, benzodiazepines), nitrosoureas (e.g., carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine), enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin γ1I and calicheamicin ω1I (Angew. Chem. Intl. Ed. Engl. 33:183-186, 1994)), dynemicins (including dynemicin A), bisphosphonates (e.g., clodronate), esperamicin , and neocarzinostatin chromophores and related chromoprotein enediyne antibiotic chromophores), antibiotics such as aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxol bicin, and deoxydoxorubicin), mitomycins such as epirubicin, esorubicin, idarubicin, marcelomycin, and mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, chelamycin, rhodrubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, antimetabolites (such as methotrexate and 5-fluorouracil (5-FU)), folic acid analogs (denopterin, methotrexate, pteropterin,Trimetrexate, etc.), purine analogues (fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, etc.), pyrimidine analogues (ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, etc.), androgens (calusterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, etc.), antiadrenal agents (aminoglutethimide, mitotane, trilostane, etc.), folic acid replacement fluids (furoic acid, etc.), aceglatone, aldophosphamide glycoside, a Minolevulinic acid, eniluracil, amsacrine, bestravcil, bisantrene, edatrexate, defofamine, demecolcine, diaziquone, elfomitin, elliptinium acetate, epothilone, etoglucide, gallium nitrate, hydroxyurea, lentinan, lonidynin, maytansinoids (such as maytansine and ansamitocin), mitoguazone, mitoxantrone, mopidamol, nitraelin, pentostatin, phenamet, pirarubicin, losoxantrone, podophyllic acid, 2-ethylhydrazide, procarbazine, PSK® polysaccharide complex (JHS) Natural Products, Eugene, Oreg.), razoxane, rhizoxin, sizofiran, spirogermanium, tenuazonic acid, triaziquone, 2,2',2''-trichlorotriethylamine, trichothecenes (especially T-2 toxin, veracrine A, roridin A, and anguidine), urethane, vindesine, dacarbazine, mannomustine, mitobronitol, mitolactol, pipobroman, gacytosine, arabinoside ("Ara-C"), cyclophosphamide, thiotepa, taxoids such as TAXOL (paclitaxel; Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE® (cremophor-free), albumin-engineered nanoparticle formulations of paclitaxel (American Pharmaceutical Partners, Schaumberg, III.), and TAXOTERE® (docetaxel; Sanofi-Aventis), chlorambucil, GEMZAR® (gemcitabine),6-thioguanine, mercaptopurine, methotrexate, platinum analogs (such as cisplatin and carboplatin), vinblastine, etoposide (VP-16), ifosfamide, mitoxantrone, vincristine, NAVELBINE® (vinorelbine), novantrone, teniposide, edatrexate, daunomycin, aminopterin, capecitabine (XELODA®), ibandronate, CPT-11, the topoisomerase inhibitor RFS2000, difluoromethylornithine (DMFO), retinoids (such as retinoic acid), and pharmaceutically acceptable salts, acids, and derivatives of any of the above.

[0162] Chemotherapeutic agents include antihormonal agents that act to regulate or inhibit hormone action on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs) (e.g., tamoxifen (NOLVADEX®, including tamoxifen citrate), raloxifene, droloxifene, iodoxifene, 4-hydroxytamoxifen, trioxifene, ketoxifene, LY117018, onapristone, and FARESTON® (including toremifene citrate), which inhibit estrogen in the adrenal glands). Aromatase inhibitors (e.g., 4(5)-imidazole, aminoglutethimide, MEGASE® (megestrol acetate), AROMASIN® (exemestane, Pfizer), formestany, fadrozole, RIVISOR® (vorozole), FEMARA® (letrozole, Novartis), and ARIMIDEX® (anastrozole, AstraZeneca)) that block the aromatase enzyme that controls its production; antiandrogens (flutamide, nilutamide, bicalcin, etc.); anti-inflammatory drugs, such as thiazolamide, leuprolide, and goserelin), buserelin, triptorelin, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, all-trans retinoic acid, fenretinide, and troxacitabine (a 1,3-dioxolane nucleoside cytosine analog), protein kinase inhibitors, lipid kinase inhibitors, antisense oligonucleotides, especially those that inhibit the expression of genes in signal transduction pathways involved in abnormal cell growth (e.g., PKC-alpha, Ralf , and H-Ras), ribozymes (VEGF expression inhibitors (e.g., ANGIOZYME® and HER2 expression inhibitors), vaccines such as gene therapy vaccines (e.g., ALLOVECTIN®, LEUVECTIN®, and VAXID®, PROLEUKIN®, rIL-2), topoisomerase 1 inhibitors (e.g., LURTOTECAN®), ABARELIX® rmRH, and pharmaceutically acceptable salts, acids, and derivatives of any of the above.

[0163] Chemotherapeutic agents also 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 the antibody-drug conjugate, gemtuzumab ozogamicin (MYLOTARG®, Wyeth). Additional humanized monoclonal antibodies with therapeutic potential as agents in combination with the compounds of the invention include apolizumab, aselizumab, atlizumab, bapineuzumab, bivatuzumab mertansine, cantuzumab mertansine, cedelizumab, certolizumab pegol, cidfusituzumab, cidtuzumab, daclizumab, eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab, fontolizumab, gemtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, labetuzumab, lintuzumab, matuzumab, mepolizumab, motavizumab, motavizumab, natalizumab, nimotuzumab, norobizumab, numab, Vizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pecfusituzumab, pectuzumab, pexelizumab, ralivizumab, ranibizumab, reslivizumab, reslizumab, resivizumab, rovelizumab, ruplizumab, sibrotuzumab, siplizumab, sontuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tefibazumab, tocilizumab, toralizumab, tucotuzumab celmoleukin, tuxituzumab, umavizumab, urtoxazumab, ustekinumab, visilizumab, and exclusively human sequence recombinant full-length IgG1 genetically modified to recognize the interleukin-12p40 protein. and the lambda antibody, anti-interleukin-12 (ABT-874 / J695, Wyeth Research and Abbott Laboratories).

[0164] Chemotherapeutic agents also include "EGFR inhibitors," alternatively referred to as "EGFR antagonists," which refer to compounds that bind to or otherwise directly interact with EGFR and disrupt or reduce its signaling activity. 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 CRL 8508), MAb528 (ATCC CRL 8509) (see U.S. Pat. No. 4,943,533, Mendelsohn et al.), and variants thereof, such as chimerized 225 (C225 or cetuximab, ERBUTIX®) and reconstituted human 225 (H225) (WO 96 / 40210, Imclone Systems, Inc.). Inc.), IMC-11F8, a fully human EGFR-targeting antibody (Imclone), antibodies that bind to type II mutant EGFR (U.S. Pat. No. 5,212,290), humanized and chimeric antibodies that bind to EGFR as described in U.S. Pat. No. 5,891,996, and human antibodies that bind to EGFR, such as ABX-EGF or panitumumab (WO 98 / 50433, Abgenix / Amgen), EMD55900 (Stragliotto et al., Eur. J. Can. cer32A:636-640 (1996)), EMD7200 (matuzumab) (a humanized EGFR antibody against EGFR that competes for EGFR binding of both EGF and TGF-alpha (EMD / Merck), human EGFR antibodies, HuMax-EGFR (GenMab), fully human antibodies known as E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3, and E7.6.3, and described in U.S. Pat. No. 6,235,883, MDX-447 (Medarex Inc), and mAb806 or humanized mAb806 (Johns et al., J. Biol. Chem. 279(29):30375-30384 (2004)). Anti-EGFR antibodies can be conjugated to cytotoxic agents to generate immunoconjugates (see, e.g., EP 659,439 A2, Merck Patent GmbH).EGFR antagonists are disclosed 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, 5,800,000, and 5,810,000. and 5,747,498, and the following PCT publications: WO98 / 14451, WO98 / 50038, WO99 / 09016, and WO99 / 24037.Specific small molecule EGFR antagonists include OSI-774 (CP-358774, erlotinib, TARCEVA®, 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®) 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), 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-quinolinyl]-4-(dimethylamino)-2-butynamide) (Wyeth), AG1478 (Pfizer), AG1571 (SU5271, Pfizer), and dual EGFR / HER2 tyrosine kinase inhibitors such as lapatinib (TYKERB®, GSK572016 or N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6[5[[[2methylsulfonyl)ethyl]amino]methyl]-2-furanyl]-4-quinazolinamine).

[0165] Chemotherapeutic agents also include "tyrosine kinase inhibitors" (including the EGFR-targeted drugs described in the previous paragraph), 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) which preferentially binds to EGFR but inhibits both HER2 and EGFR overexpressing cells), lapatinib (GSK572016, available from Glaxo-SmithKline), oral HER2 and EGFR tyrosine kinase inhibitor, PKI-166 (available from Novartis), pan-HER inhibitors (such as canertinib (CI-1033, Pharmacia)), Raf-1 inhibitors (such as ISIS, which inhibits Raf-1 signaling) antisense drug ISIS-5132 available from Novartis / Schering Pharmaceuticals), non-HER-targeted TK inhibitors (such as imatinib mesylate (GLEEVEC®, available from GlaxoSmithKline)), multi-targeted tyrosine kinase inhibitors (such as sunitinib (SUTENT®, available from Pfizer)), VEGF receptor tyrosine kinase inhibitors (vatalanib (PTK787 / ZK222584, Novartis / Schering)), available from AG), MAPK extracellular regulated kinase I inhibitor CI-1040 (available from Pharmacia), quinazolines (such as PD153035, 4-(3-chloroanilino)quinazoline), pyridopyrimidines, pyrimidopyrimidines, pyrrolopyrimidines (such as CGP59326, CGP60261, and CGP62706), pyrazolopyrimidines, 4-(phenylamino)-7H-pyrrolo[2,3-d]pyrimidine, curcumin (diferuloylmethane, 4,5-bis(4-fluoroanilino)phthalimide), tyrphostins containing a nitrothiophene moiety, PD-0183805 (Warner-Lambert), antisense molecules (e.g., those that bind to HER-encoding nucleic acids), quinoxalines (U.S. Pat. No. 5,804,396), tryphostins (U.S. Pat. No. 5,804,No. 396), ZD6474 (AstraZeneca), PTK-787 (Novartis / Schering AG), pan-HER inhibitors (such as CI-1033 (Pfizer)), Affinitac (ISIS3521, Isis / Lilly), imatinib mesylate (GLEEVEC®), PKI166 (Novartis), GW2016 (GlaxoSmithKline), 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. Pat. No. 5,804,396, WO1999 / 09016, WO1998 / 43960, WO1997 / 38983, WO1999 / 06378, WO1999 / 06396, WO1996 / 30347, WO1996 / 33978, WO1996 / 3397, and WO1996 / 33980.

[0166] The term "multi-targeted tyrosine kinase inhibitor" as used herein refers to a tyrosine kinase inhibitor that inhibits multiple (i.e., two or more) tyrosine kinase proteins. The tyrosine kinase proteins may be receptor tyrosine kinases and / or cellular tyrosine kinases. For example, the multi-targeted tyrosine kinase inhibitor may inhibit platelet-derived growth factor receptors (e.g., PDGFR-αα, PDGFR-ββ, and / or PDGFR-αβ), VEGF receptors (e.g., VEGFR1 and / or VEGFR2), CD117 (c-Kit), RET, CD114, and / or CD135. Exemplary multi-targeted tyrosine kinase inhibitors include sunitinib (N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-1,2-dihydro-3H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-carboxamide, also known as SUTENT®, or SU11248), SU6656, motesanib, sorafenib (e.g., NEXEVAR® or BAY439006), axitinib, afatinib, bosutinib, crizotinib, cabozantinib, dasatinib, entrectinib, pazopanib, lapatinib, and vandetanib (also known as ZACTIMA® or ZD6474). It should be understood that a multi-targeted tyrosine kinase inhibitor that inhibits a VEGF receptor can also be considered a VEGFR inhibitor.

[0167] Chemotherapeutic agents also include dexamethasone, interferon, colchicine, metoprine, cyclosporine, amphotericin, metronidazole, alemtuzumab, alitretinoin, allopurinol, amifostine, arsenic trioxide, asparaginase, live BCG, bevacuzimab, bexarotene, cladribine, clofarabine, darbepoetin alfa, denileukin, dexrazoxane, epoetin alfa, erlotinib, filgrastim, histrelin acetate, ibritumomab, interferon alfa-2a, interferon alfa-2b, lenalid Also included are acetaminophen, levamisole, mesna, methoxsalen, nandrolone, nelarabine, nofetumomab, oprelvekin, palifermin, pamidronate, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium, quinacrine, rasburicase, sargramostim, temozolomide, VM-26, 6-TG, toremifene, tretinoin, all-trans retinoic acid (ATRA), valrubicin, zoledronate, and zoledronic acid, and pharmaceutically acceptable salts thereof.

[0168] The term "prodrug" as used herein refers to a precursor or derivative form of a pharmaceutically active substance that has reduced cytotoxicity to tumor cells compared to the parent drug and can be enzymatically activated or converted into a more active parent form.See, for example, Wilman, "Prodrugs in Cancer Chemotherapy," Biochemical Society Transactions, 14, pp. 375-382, 615th Meeting Belfast (1986) and Stella et al., "Prodrugs: A Chemical Approach to Targeted Drug Delivery," Directed Drug Delivery, Borchardt et al. (eds.), pp. 247-267, Humana Press (1985). The prodrugs of the present invention include, but are not limited to, phosphate-containing prodrugs, thiophosphate-containing prodrugs, sulfate-containing prodrugs, peptide-containing prodrugs, D-amino acid modified prodrugs, glycosylated prodrugs, β-lactam-containing prodrugs, optionally substituted phenoxyacetamide-containing prodrugs, or optionally substituted phenylacetamide-containing prodrugs, 5-fluorocytosine and other 5-fluorouridine prodrugs, which can be converted into more active cytotoxic free drugs. Examples of cytotoxic drugs that can be derivatized into prodrug forms for use in the present invention include, but are not limited to, the chemotherapeutic agents described above.

[0169] A "growth inhibitory agent," as used herein, refers to a compound or composition that inhibits the growth and / or proliferation of cells (e.g., cells whose growth is dependent on PD-L1 expression) either in vitro or in vivo. Thus, a growth inhibitory agent may be one that significantly reduces the percentage of cells in S phase. Examples of growth inhibitory agents include agents that block cell cycle progression (at a place other than S phase), such as agents that induce G1 arrest and M-phase arrest. Standard M-phase blockers include vincas (vincristine and vinblastine), taxanes, and topoisomerase II inhibitors, such as the anthracycline antibiotic doxorubicin ((8S-cis)-10-[(3-amino-2,3,6-trideoxy-α-L-lyxo-hexapyranosyl)oxy]-7,8,9,10-tetrahydro-6,8,11-trihydroxy-8-(hydroxyacetyl)-1-methoxy-5,12-naphthacenedione), epirubicin, daunorubicin, etoposide, and bleomycin. Agents that arrest G1 also extend to S-phase arrest, such as DNA alkylating agents, such as tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C. Further information can be found in "The Molecular Basis of Cancer" by Murakami et al., edited by Mendelsohn and Israel, Chapter 1, entitled "Cell cycle regulation, oncogenes, and antitineoplastic drugs" (WB Saunders: Philadelphia, 1995), particularly page 13. Taxanes (paclitaxel and docetaxel) are anticancer drugs, both derived from the yew tree. Docetaxel (TAXOTERE®, Rhone-Poulenc Rorer), derived from the European yew, is a semisynthetic analog of paclitaxel (TAXOL®, Bristol-Myers Squibb). Paclitaxel and docetaxel promote the assembly of microtubules derived from tubulin dimers, stabilize microtubules by preventing depolymerization, and inhibit mitosis in cells.

[0170] "Radiation therapy" refers to the use of directed gamma or beta radiation to induce sufficient damage to cells so as to limit their ability to function normally or to destroy them entirely. It will be appreciated that there are many methods known in the art for determining dose and duration of treatment. A typical treatment is given as a single administration, with typical doses ranging from 10 to 200 units (Gy) per day.

[0171] The term "pharmaceutical formulation" refers to a preparation in which the biological activity of the active ingredient contained therein is in a form such that it is effective, and which does not contain any additional components that are unacceptably toxic to the patient to whom the formulation is administered.

[0172] A "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation, other than an active ingredient, that is non-toxic to a patient. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0173] The term "package insert" is used to refer to instructions typically included in commercial packaging of a therapeutic product that contain information about directions, usage, dosage, administration, concomitant therapy, contraindications, and / or warnings regarding the use of such therapeutic product.

[0174] A "sterile" preparation is aseptic or free of all viable microorganisms and their spores.

[0175] An "article of manufacture" is any manufactured article (e.g., package or container) or kit that includes at least one reagent, e.g., a pharmaceutical agent for the treatment of a disease or disorder (e.g., cancer), or a probe for specifically detecting a biomarker described herein. In certain embodiments, the article of manufacture or kit is promoted, distributed, or sold as a unit for performing a method described herein.

[0176] The term "small molecule" refers to any molecule having a molecular weight below about 2000 daltons, preferably below about 500 daltons.

[0177] The term "label" as used herein refers to a compound or composition that is directly or indirectly conjugated or fused to a reagent, such as a polynucleotide probe or antibody, to facilitate detection of the conjugated or fused reagent. The label may be detectable itself (e.g., a radioisotope label or a fluorescent label), or, in the case of an enzymatic label, may catalyze the chemical modification of a detectable substrate compound or composition. This term is intended to encompass both direct labeling of a probe or antibody by binding (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of a probe or antibody by reactivity with another directly labeled reagent. Examples of indirect labeling include detection of a primary antibody using a fluorescently labeled secondary antibody and end-labeling of a DNA probe with biotin so that it can be detected with fluorescently labeled streptavidin.

[0178] The term "antibody" is used in the broadest sense and specifically covers monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired biological activity.

[0179] "Native antibodies" are typically heterotetrameric glycoproteins of approximately 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide bonds varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced interchain disulfide bridges. Each heavy chain has a variable domain (VH) at one end followed by several constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end, with the light chain constant domain aligned with the first constant domain of the heavy chain and the light chain variable domain aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light chain variable domain and the heavy chain variable domain.

[0180] An "isolated" antibody is one that has been identified and separated and / or recovered from a component of its natural environment. Contaminant components of its natural environment are substances that would interfere with research, diagnostic, and / or therapeutic uses of the antibody, including enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In some embodiments, the antibody is purified (1) to greater than 95% by weight, and in some embodiments, greater than 99% by weight, as determined, for example, by the Lowry method; (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence, for example, using a spinning cup sequenator; or (3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions, for example, using Coomassie blue or silver staining. Isolated antibodies include antibodies in situ within recombinant cells, since at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated antibodies will be prepared by at least one purification step.

[0181] A "blocking" antibody or antibody "antagonist" is one that inhibits or reduces the biological activity of the antigen to which it binds. For example, a VEGF-specific antagonist antibody binds to VEGF and inhibits the ability of VEGF to induce vascular endothelial cell proliferation. Preferred blocking or antagonist antibodies completely inhibit the biological activity of the antigen.

[0182] Unless otherwise indicated, the phrase "multivalent antibody" is used throughout this specification to refer to an antibody that comprises three or more antigen-binding sites. A multivalent antibody is preferably engineered to have three or more antigen-binding sites and is generally not a native sequence IgM or IgA antibody.

[0183] The "light chains" of antibodies (immunoglobulins) from any mammalian species can be assigned to one of two clearly distinct types, called kappa ("κ") and lambda ("λ"), based on the amino acid sequences of their constant domains.

[0184] The term "constant domain" refers to the portion of an immunoglobulin molecule that has a more conserved amino acid sequence compared to the other portion of the immunoglobulin, the variable domain, which contains the antigen-binding site. The constant domain includes the CH1, CH2, and CH3 domains of the heavy chain (collectively, CH), and the CHL (or CL) domain of the light chain.

[0185] 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 domain of the heavy chain may be referred to as "VH." The variable domain of the light chain may be referred to as "VL." These domains are generally the most variable parts of an antibody and contain the antigen-binding site.

[0186] The term "variable" refers to the fact that certain segments of the variable domains differ extensively in sequence among antibodies. The variable or "V" domains mediate antigen binding and define the specificity of a particular antibody for its particular antigen. However, variability is not evenly distributed across the span of the variable domains. Instead, V regions consist of relatively invariant stretches of 15-30 amino acids called framework regions (FRs), separated by short regions of extreme variability called "hypervariable regions," each 9-12 amino acids long. The term "hypervariable region" or "HVR," as used herein, refers to the amino acid residues of an antibody responsible for antigen binding. The hypervariable regions generally comprise, for example, amino acid residues from about residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the VL, and from about residues 26-35 (H1), 49-65 (H2), and 95-102 (H3) in the VH (in one embodiment, H1 is from about residues 31-35) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. (1991)), and / or residues from the "hypervariable loops" (e.g., residues 26-32 (L1), 50-52 (L2), and 91-96 (L3) in VL, and residues 26-32 (H1), 53-55 (H2), and 96-101 (H3) in VH (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). Native heavy and light chain variable domains each contain four FRs that predominantly adopt a beta-sheet configuration connected by three hypervariable regions, which form loops that connect, and in some cases, form part of, the beta-sheet structure.The hypervariable regions in each chain are held in close proximity by FRs, which, together with the hypervariable regions from the other chain, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, (1991)). Thus, the HVR and FR sequences generally follow the sequence FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4 in VH (or VL). The constant domains are not directly involved in binding an antibody to an antigen, but exhibit various effector functions, such as antibody-dependent cellular cytotoxicity (ADCC).

[0187] For purposes herein, an "acceptor human framework" is a framework comprising the amino acid sequence of a light chain variable domain (VL) framework or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human consensus framework, as defined below. An acceptor human framework "derived from" a human immunoglobulin framework or a human consensus framework may comprise the same amino acid sequence or may contain amino acid sequence changes. In some embodiments, the number of amino acid changes is 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 or fewer, 4 or fewer, 3 or fewer, or 2 or fewer. In some embodiments, the VL acceptor human framework is identical in sequence to the VL human immunoglobulin framework sequence or the human consensus framework sequence.

[0188] The terms "hypervariable region," "HVR," or "HV" as used herein refer to the region of an antibody variable domain that is hypervariable in sequence and / or forms structurally defined loops. Generally, antibodies contain six HVRs: three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). In natural antibodies, H3 and L3 exhibit the highest diversity of the six HVRs, and H3 in particular is thought to play a unique role in conferring superior specificity to antibodies. See, for example, Xu et al., Immunity 13:37-45 (2000); Johnson and Wu, Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, NJ, 2003). In fact, naturally occurring camelid antibodies, consisting only of heavy chains, are functional and stable in the absence of light chains. See, for example, Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al., Nature Struct. Biol. 3:733-736 (1996).

[0189] Several HVR delineations are used and encompassed herein. Kabat complementarity-determining regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). Chothia instead refers to the location of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). AbM HVRs represent a compromise between Kabat HVRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. "Contact" HVRs are based on analysis of available complex crystal structures. Residues from each of these HVRs are shown below. Loop Kabat AbM Chothia Contact L1 L24-L34 L24-L34 L26-L32 L30-L36 L2 L50-L56 L50-L56 L50-L52 L46-L55 L3 L89-L97 L89-L97 L91-L96 L89-L96 H1 H31-H35b H26-H35b H26-H32 H30-H35b (Kabat numbering) H1 H31-H35 H26-H35 H26-H32 H30-H35 (Chothia numbering) H2 H50-H65 H50-H58 H53-H55 H47-H58 H3 H95-H102 H95-H102 H96-H101 H93-H101

[0190] HVRs may include the following "extended HVRs": 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL, and 26-35 (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) in VH. The variable domain residues are numbered according to Kabat et al. (see above) for each of these definitions.

[0191] "Framework" or "FR" residues are those variable domain residues other than the HVR residues as herein defined.

[0192] A "human consensus framework" is a framework representing the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. Generally, the subgroup of sequences is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., NIH Publication 91-3242, Bethesda, MD (1991), vols. 1-3. In one embodiment, for VL, the subgroup is subgroup kappa I as in Kabat et al. (supra). In one embodiment, for VH, the subgroup is subgroup III as in Kabat et al. (supra).

[0193] The terms "variable domain residue numbering as in Kabat" or "amino acid position numbering as in Kabat," and variations thereof, refer to the numbering system used for the heavy or light chain variable domains of the compilation of antibodies in Kabat et al. (see above). Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening of, or insertion into, the FRs or HVRs of the variable domain. For example, a heavy chain variable domain may contain a single amino acid insertion after residue 52 of H2 (residue 52a according to Kabat) and inserted residues after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat, etc.). The Kabat numbering of residues can be determined for a given antibody by alignment of the antibody's sequence with the "standard" Kabat-numbered sequence at the regions of homology.

[0194] The Kabat numbering system is generally used when referring to residues within the variable domain (approximately residues 1-107 of the light chain and residues 1-113 of the heavy chain) (e.g., Kabat et al., Sequences of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The "EU numbering system" or "EU index" is generally used when referring to residues in immunoglobulin heavy chain constant regions (e.g., the EU index reported in Kabat et al., supra). The "EU index as in Kabat" refers to the residue numbering of the human IgG1 EU antibody. Unless otherwise indicated herein, references to residue numbers in the variable domain of an antibody refer to residue numbering according to the Kabat numbering system. Unless otherwise indicated herein, references to residue numbers in the constant domain of an antibody refer to residue numbering according to the EU numbering system (see, e.g., U.S. Provisional Application No. 60 / 640,323, figures relating to EU numbering).

[0195] Unless otherwise indicated, HVR residues and other residues in the variable domain (e.g., FR residues) are numbered herein according to Kabat et al. (supra).

[0196] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody in its substantially intact form, rather than an antibody fragment, as defined below. These terms specifically refer to an antibody having a heavy chain that includes an Fc region.

[0197] An "antibody fragment" comprises a portion of an intact antibody, preferably comprising its antigen-binding region. In some embodiments, the antibody fragments described herein are antigen-binding fragments. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments, diabodies, linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.

[0198] Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment, named for its ability to crystallize readily. Pepsin treatment yields an F(ab')2 fragment, which has two antigen-binding sites and is still capable of cross-linking antigen.

[0199] The term "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain containing at least a portion of the constant region. This term includes native-sequence Fc regions and variant Fc regions. In one embodiment, a human IgG heavy chain Fc region extends from Cys226, or from Pro230, to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region follows the EU numbering system (also referred to as the EU index), as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991).

[0200] "Effector function" refers to a biological activity attributable to the Fc region of an antibody, which varies depending on the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0201] An "Fv" is the minimum antibody fragment that contains a complete antigen-binding site. In one embodiment, a two-chain Fv species consists of a dimer of one heavy-chain variable domain and one light-chain variable domain in tight, non-covalent association. In a single-chain Fv (scFv) species, one heavy-chain variable domain and one light-chain variable domain can be covalently linked by a flexible peptide linker so that the light and heavy chains can associate in a "dimeric" structure similar to that in a two-chain Fv species. It is in this configuration that the three HVRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six HVRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three antigen-specific HVRs) has the ability to recognize and bind to antigen, albeit with lower affinity than the entire binding site.

[0202] Fab fragments contain heavy and light chain variable domains, and also contain a light chain constant domain and the first heavy chain constant domain (CH1). Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine ​​residue(s) of the constant domains bear a free thiol group. F(ab')2 antibody fragments were originally produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0203] "Single-chain Fv" or "scFv" antibody fragments comprise the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Generally, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains, which enables the scFv to form the desired structure for antigen binding. For a review of scFvs, see, for example, Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, edited by Rosenburg and Moore (Springer-Verlag, New York, 1994), pp. 269-315.

[0204] The term "multispecific antibody" is used in the broadest sense and specifically encompasses antibodies comprising a heavy chain variable domain (VH) and a light chain variable domain (VL) in which the VH-VL unit has polyepitopic specificity (i.e., can bind to two different epitopes on one biological molecule, or each epitope on a different biological molecule). Such multispecific antibodies include, but are not limited to, full-length antibodies, antibodies having two or more VL and VH domains, antibody fragments such as Fab, Fv, dsFv, scFv, diabodies, bispecific diabodies, and triabodies, as well as covalently or noncovalently linked antibody fragments. "Polyepitopic specificity" refers to the ability to specifically bind to two or more different epitopes on the same or different target(s). "Bispecificity" or "dual specificity" refers to the ability to specifically bind to two different epitopes on the same or different target(s). However, in contrast to bispecific antibodies, diabodies have two antigen-binding arms with identical amino acid sequences, and each Fab arm can recognize two antigens. Bispecificity allows an antibody to interact with two different antigens as a single Fab or IgG molecule with high affinity. According to one embodiment, a multispecific antibody in IgG1 binds to each epitope with an affinity of 5 μM to 0.001 pM, 3 μM to 0.001 pM, 1 μM to 0.001 pM, 0.5 μM to 0.001 pM, or 0.1 μM to 0.001 pM. "Monospecificity" refers to the ability to bind to only one epitope.

[0205] The term "diabody" refers to an antibody fragment having two antigen-binding sites, comprising a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, these domains are forced to pair with complementary domains on another chain, creating two antigen-binding sites. Diabodies can be bivalent or bispecific. Diabodies are more fully described, for example, in EP 404,097, WO 1993 / 01161, Hudson et al., Nat. Med. 9:129-134 (2003), and Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003).

[0206] The "class" of an antibody refers to the type of constant domain or constant region carried by its heavy chain. There are five major classes of antibodies (IgA, IgD, IgE, IgG, and IgM), some of which can be further divided into "subclasses" (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of antibodies are called α, δ, ε, γ, and μ, respectively.

[0207] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible mutations that may be present in minor amounts, e.g., naturally occurring mutations. Thus, the modifier "monoclonal" indicates the character of the antibody as not being a mixture of distinct antibodies. In certain embodiments, such monoclonal antibodies typically comprise an antibody comprising a polypeptide sequence that binds to a target, the target-binding polypeptide sequence being obtained by a process that includes selecting a single target-binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, or recombinant DNA clones. It should be understood that the selected target-binding sequence may be further modified, for example, to improve affinity for the target, humanize the target-binding sequence, improve its production in cell culture, reduce its immunogenicity in vivo, create a multispecific antibody, etc., and that an antibody comprising the modified target-binding sequence is also a monoclonal antibody of the present invention. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are typically uncontaminated by other immunoglobulins.

[0208] The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention can be produced using, for example, hybridoma techniques (e.g., Kohler and Milstein, Nature 256:495-97 (1975); Hongo et al., Hybridoma 14(3):253-260 (1995); Harlow et al., Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 2nd ed., 1988); Hammerling et al., Monoclonal Antibodies and T Cell Hybridomas 563-681 (Elsevier, NY, 1981)), recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567), phage display technology (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 the like. , and Lee et al., J. Immunol. Methods 284(1-2):119-132, 2004), and techniques for producing human or human-like antibodies in animals that have some or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (e.g., WO 1998 / 24893, WO 1996 / 34096, WO 1996 / 33735, WO 1991 / 10741, Jakobovits et al., Proc. Natl. Acad. Sci. USA 90:2551, 1993, Jakobovits et al., Nature 362:255-258, 1993, Bruggemann et al., Year in Immunol.7:33, 1993; U.S. Patent Nos. 5,545,807, 5,545,806, 5,569,825, 5,625,126, 5,633,425, and 5,661,016; Marks et al., Bio / Technology 10:779-783 (1992); Lonberg et al., Nature 368:856-859, 1994; Morrison, Nature 368:812-813, 1994; Fishwild et al., Nature Biotechnol. 14:845-851, 1996; Neuberger, Nature Biotechnol. 14:826, 1996, and Lonberg et al. Intern. Rev. Immunol. 13:65-93, 1995).

[0209] The monoclonal antibodies herein specifically include "chimeric" antibodies in which a portion of the heavy and / or light chain is identical to or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical to or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (see, e.g., U.S. Pat. No. 4,816,567 and Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)). Chimeric antibodies include PRIMATIZED® antibodies, whose antigen-binding regions are derived from, for example, antibodies produced by immunizing macaque monkeys with an antigen of interest.

[0210] A "human antibody" is one that possesses an amino acid sequence that corresponds to that of an antibody produced by a human or human cell, or derived from a non-human source that utilizes the human antibody repertoire or other human antibody coding sequences. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues.

[0211] "Humanized" forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human primate having the desired antibody specificity, affinity, and capacity. In some instances, FR residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or the donor antibody. These modifications are made to further refine antibody performance. In general, humanized antibodies will comprise substantially all of at least one, and typically two, variable domains, with all or substantially all of the hypervariable loops corresponding to those of a non-human immunoglobulin and all or substantially all of the FRs being those of a human immunoglobulin sequence. Humanized antibodies will also optionally comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see 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.

[0212] A "wild-type (WT)" or "reference" sequence, or the sequence of a "wild-type" or "reference" protein / polypeptide (such as an HVR or variable domain of a reference antibody), can be a reference sequence from which variant polypeptides are derived through the introduction of mutations. Generally, the "wild-type" sequence of a given protein is the most common sequence found in nature. Similarly, a "wild-type" gene sequence is the sequence for that gene that is most commonly found in nature. Mutations can be introduced into a "wild-type" gene (and thus the protein it encodes) through natural processes or through artificial means. The product of such a process is a "variant" or "mutant" form of the original "wild-type" protein or gene.

[0213] A "variant" or "mutant" of a starting or reference polypeptide (e.g., a reference antibody or variable domain(s) / HVR(s) thereof) is a polypeptide that (1) has an amino acid sequence that differs from that of the starting or reference polypeptide and (2) is derived from the starting or reference polypeptide through either natural or artificial mutagenesis. Such variants include, for example, deletions from, and / or insertions into, and / or substitutions of residues within the amino acid sequence of a polypeptide of interest, referred to herein as "amino acid residue modifications." Thus, a variant HVR refers to an HVR that comprises a variant sequence with respect to a starting or reference polypeptide sequence (such as that of a source antibody or antigen-binding fragment). An amino acid residue modification, in this context, refers to an amino acid that differs from the amino acid at the corresponding position in the starting or reference polypeptide sequence (such as that of a reference antibody or fragment thereof). Any combination of deletion, insertion, and substitution may be used to arrive at the final variant or mutant construct, provided that the final construct possesses the desired functional characteristics. Amino acid changes may also alter post-translational processing of the polypeptide, such as changing the number or position of glycosylation sites.

[0214] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described herein.

[0215] With respect to the binding of an antibody to a target molecule, the term "specific binding" or "specifically binds" or "specifically" to a particular polypeptide or epitope on a particular polypeptide target refers to binding that is measurably different from non-specific interactions. Specific binding can be measured, for example, by determining the binding of a molecule in comparison to the binding of a control molecule. For example, specific binding can be determined by competition with a control molecule that is similar to the target, e.g., excess unlabeled target. In this case, specific binding is indicated when the binding of the labeled target to the probe is competitively inhibited by excess unlabeled target. As used herein, the term "specific binding" or "specifically binds" or "specifically" to a particular polypeptide or epitope on a particular polypeptide target refers to binding that is measurably different from non-specific interactions, for example, when the binding of a control molecule is competitively inhibited by excess unlabeled target. -4 M or less, or 10 -5 M or less, or 10 -6 M or less, or 10 -7 M or less, or 10 -8 M or less, or 10 -9 M or less, or 10 -10 M or less, or 10 -11 M or less, or 10 -12 M or less, or 10 -4 M~10 -6 M or 10 -6 M~10 -10 M or 10 -7 M~10 -9 Affinity can be exhibited by a molecule having a Kd in the range of M. As will be understood by those skilled in the art, affinity and Kd values ​​are inversely related. A high affinity for an antigen is measured by a low Kd value. In one embodiment, the term "specific binding" refers to binding when a molecule binds to a particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptides or polypeptide epitopes.

[0216] An "affinity matured" antibody refers to an antibody that has one or more alterations in one or more hypervariable regions (HVRs) compared to a parent antibody that does not possess such alterations, which improve the affinity of the antibody for its antigen.

[0217] An "antibody that binds to the same epitope" as a reference antibody refers to an antibody that blocks the reference antibody from binding to its antigen by 50% or more in a competitive assay, and conversely, a reference antibody that blocks the antibody from binding to its antigen by 50% or more in a competitive assay.

[0218] An "immunoconjugate" is an antibody conjugated to one or more heterologous molecule(s), including, but not limited to, a cytotoxic agent.

[0219] As used herein, the term "immunoadhesin" refers to an antibody-like molecule that combines the binding specificity of a heterologous protein ("adhesin") with the effector functions of immunoglobulin constant domains. Structurally, immunoadhesins comprise a fusion of an amino acid sequence with the desired binding specificity, other than the antigen recognition and binding site of an antibody (i.e., "heterologous"), with an immunoglobulin constant domain sequence. The adhesin portion of an immunoadhesin molecule is typically a contiguous amino acid sequence that includes at least the binding site of a receptor or ligand. The immunoglobulin constant domain sequence in an immunoadhesin can be derived from any immunoglobulin, such as IgG1, IgG2 (including IgG2A and IgG2B), IgG3, or IgG4 subtypes, IgA (including IgA1 and IgA2), IgE, IgD, or IgM. Ig fusions preferably involve the substitution of a polypeptide or antibody domain as described herein in place of at least one variable region within an Ig molecule. In a particularly preferred embodiment, the immunoglobulin fusion comprises the hinge, CH2, and CH3 regions, or the hinge, CH1, CH2, and CH3 regions, of an IgG1 molecule. See also U.S. Patent No. 5,428,130 for information on the generation of immunoglobulin fusions. For example, immunoadhesins useful as therapeutic medicaments herein include polypeptides comprising the extracellular domain (ECD) or PD-1-binding portion of PD-L1 or PD-L2, or the extracellular or PD-L1- or PD-L2-binding portion of PD-1 (such as PD-L1 ECD-Fc, PD-L2 ECD-Fc, and PD-1 ECD-Fc, respectively), fused to a constant domain of an immunoglobulin sequence. Immunoadhesin combinations of the Ig Fc and ECD of cell surface receptors are sometimes referred to as soluble receptors.

[0220] "Fusion protein" and "fusion polypeptide" refer to a polypeptide having two covalently linked portions, each of which is a polypeptide with a different property. This property can be a biological property, such as in vitro or in vivo activity. This property can also be a simple chemical or physical property, such as binding to a target molecule or catalysis of a reaction. The two portions can be linked directly by a single peptide bond or through a peptide linker, but they are in each other's reading frame.

[0221] "Percent (%) amino acid sequence identity" with respect to the polypeptide sequences identified herein is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the polypeptide being compared, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in a variety of ways within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared. However, for purposes herein, 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, together with user documentation, has been filed with the U.S. Copyright Office, Washington, DC 20559, and is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California. The ALIGN-2 program must be compiled for use on a UNIX operating system, preferably Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.

[0222] In situations 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 opposite to a given amino acid sequence B (alternatively, it can be written as, "given amino acid sequence A has or contains a certain % amino acid sequence identity to, with, or opposite to sequence B" for a given amino acid sequence B) is calculated as follows: 100 x fraction X / Y

[0223] where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in a programmatic alignment of A and B, and Y is the total number of amino acid residues in B. It will be understood that if the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not be equal to the % amino acid sequence identity of B to A. Unless otherwise specified, all % amino acid sequence identity values ​​used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[0224] As used interchangeably herein, "polynucleotide" or "nucleic acid" refers to a polymer of nucleotides of any length, including DNA and RNA. The nucleotides may be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or 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, but are not limited to, single- and double-stranded DNA, DNA containing single- and double-stranded regions, single- and double-stranded RNA, and RNA containing single- and double-stranded regions, and hybrid molecules containing DNA and RNA that may contain single or, more typically, double strands, or may contain single and double-stranded regions. Additionally, 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 may be from the same molecule or from different molecules. These regions may contain all of one or more of these molecules, but more typically, only one region of some of these molecules. One of the molecules in the triple helix region is often an oligonucleotide. The term "polynucleotide" specifically includes cDNA.

[0225] A polynucleotide may comprise modified nucleotides, such as methylated nucleotides and their analogs. If present, modifications to the nucleotide structure may be imparted before or after assembly of the polymer. The sequence of nucleotides may be interrupted by non-nucleotide components. A polynucleotide may be further modified after synthesis, such as by conjugation with a label. Other types of modifications include, for example, "caps," substitution of one or more of the naturally occurring nucleotides with an analog, internucleotide modifications such as those with uncharged linkages (e.g., methylphosphonates, phosphotriesters, phosphoamidates, carbamates, etc.) and those with charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), those containing pendant moieties such as proteins (e.g., nucleases, toxins, antibodies, signal peptides, poly-L-lysine, etc.), those containing intercalating agents (e.g., acridine, psoralens, etc.), those containing chelating agents (e.g., metals, radioactive metals, boron, metal oxides, etc.), those containing alkylating agents, those with modified linkages (e.g., alpha anomeric nucleic acids, etc.), and unmodified forms of the polynucleotide(s). Additionally, any of the hydroxyl groups normally present in the sugar may be replaced, for example, with a phosphonate group, a phosphate group, protected with a standard protecting group, or activated to provide for additional linkage to an additional nucleotide, or conjugated to a solid or semi-solid support. The 5' and 3' terminal OH 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 to standard protecting groups. Polynucleotides can also contain analogous forms of ribose or deoxyribose sugars known in the art, including, for example, 2'-O-methyl-, 2'-O-allyl-, 2'-fluoro-, or 2'-azido-ribose, carbocyclic sugar analogs, α-anomeric sugars, epimeric sugars such as arabinose, xylose, or lyxose, pyranose sugars, furanose sugars, sedoheptulose, acyclic 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, but are not limited to, embodiments in which phosphate is replaced by P(O)S ("thioate"), P(S)S ("dithioate"), "(O)NR2 ("amidate"), P(O)R, P(O)OR', CO, or CH2 ("formacetal"), where each R or R' is independently H or substituted or unsubstituted alkyl (1-20C) (optionally containing an ether (-O-) linkage), aryl, alkenyl, cycloalkyl, cycloalkenyl, or araldyl. Not all of the linkages in a polynucleotide need be identical. The foregoing description applies to all polynucleotides referred to herein, including RNA and DNA.

[0226] " Oligonucleotide " as used herein generally refers to a short single-stranded polynucleotide that is less than about 250 nucleotides in length, but not necessarily. Oligonucleotide may be synthetic. The terms " oligonucleotide " and " polynucleotide " are not mutually exclusive. The above description of polynucleotide is equally and completely applicable to oligonucleotide.

[0227] The term "primer" refers to a single-stranded polynucleotide that is capable of hybridizing to a nucleic acid and generally provides a free 3'-OH group, thereby allowing polymerization of a complementary nucleic acid.

[0228] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," including the primary transformed cell and its progeny regardless of the number of passages. The progeny may not be completely identical in nucleic acid content to the parent cell and may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.

[0229] The term "vector," as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors as self-replicating nucleic acid structures and vectors that integrate into the genome of a host cell into which the vector is introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as "expression vectors."

[0230] An "isolated" nucleic acid molecule is a nucleic acid molecule that has been identified and separated from at least one contaminating nucleic acid molecule, which is normally associated with the natural source of the nucleic acid. An isolated nucleic acid molecule is other than the form or state in which it is found in nature. Thus, an isolated nucleic acid molecule is distinct from the nucleic acid molecule present in natural cells. However, an isolated nucleic acid molecule also includes nucleic acid molecules contained in cells that normally express an antibody, for example, when the nucleic acid molecule is in a chromosomal location different from that of natural cells.

[0231] II. Diagnostic Methods Provided herein are methods for identifying individuals having cancer (e.g., kidney cancer (e.g., renal cell carcinoma (RCC)) who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab (MPDL3280A), or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

[0232] The methods described herein include determining the presence of sarcomatoid cancer and / or using the individual's Memorial Sloan Kettering Cancer Center (MSKCC) risk score to determine whether the individual is receiving a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab (MPDL3280A), or a PD-1 The present invention is based, at least in part, on the discovery that it is possible to identify patients who are likely to benefit from anti-cancer treatments comprising binding antagonists (e.g., anti-PD-1 antibodies). The benefit may be, for example, related to improved progression-free survival (PFS), overall survival (OS), overall response rate (ORR), complete response (CR) rate, and / or disease-free rate (DFR). For example, in some cases, the benefit may be related to PFS. In other cases, the benefit may be related to OS. In still other cases, the benefit may be related to ORR. In still other cases, the benefit may be related to CR rate. In still other cases, the benefit may be related to DFR.

[0233] The methods described herein also include assaying one or more genes (e.g., CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, TAP2, VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, CD34, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-1, IL-2, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-1, IL-2, IL-1, IL-2, IL-3, IL-4, IL-1, IL-2, IL-3, IL-4, IL-5, IL-1, IL-2, IL-1, IL-2, IL-3, IL-4 ...3, IL-4, IL-1, IL- 6. The expression levels of CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and / or S100A9 are increased after treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab)), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis binding antagonist. In another aspect, the methods and assays described herein are based, at least in part, on the discovery that the expression levels of one or more genes (e.g., VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, and / or CD34) in a sample from an individual can be used to predict the therapeutic efficacy of a treatment comprising an angiogenesis inhibitor (e.g., a VEGF antagonist (e.g., a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))).In some embodiments, a sarcomatoid cancer and / or an individual's MSKCC risk score, in combination with the expression levels of one or more genes (e.g., CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, TAP2, VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, CD34, IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and / or S100A9) in a sample from the individual, can be used, for example, to identify individuals who are likely to benefit (e.g., in terms of PFS) from an anti-cancer treatment described herein, to select individuals for an anti-cancer treatment described herein, and / or to optimize the therapeutic efficacy of an anti-cancer treatment described herein.

[0234] Further provided herein are methods for selecting a treatment for an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising: selecting a treatment for an individual having cancer; and determining whether the individual having cancer is receiving a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab (MPDL3280A), or and methods for determining whether an individual having cancer is likely to respond to treatment comprising an angiogenesis inhibitor (e.g., a VEGF antagonist (e.g., a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))). Methods for predicting the responsiveness of an individual with cancer to treatments including angiogenesis inhibitors (e.g., VEGF antagonists (e.g., VEGFR inhibitors (e.g., multi-targeted tyrosine kinase inhibitors (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), VEGF antagonists (e.g., anti-VEGF antibodies (e.g., bevacizumab), or VEGFR inhibitors (e.g., multi-targeted tyrosine kinase inhibitors (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and methods for monitoring the response of individuals with cancer to treatments comprising PD-L1 axis-binding antagonists (e.g., PD-L1 binding antagonists, (e.g., anti-PD-L1 antibodies, e.g., atezolizumab (MPDL3280A), or PD-1 binding antagonists (e.g., anti-PD-1 antibodies)), as well as to treatments comprising angiogenesis inhibitors (e.g., VEGF antagonists (e.g., VEGFR inhibitors (e.g., multi-targeted tyrosine kinase inhibitors (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))).A method for monitoring the response of an individual with cancer. Any of the methods provided herein may further include administering to the individual a VEGF antagonist and a PD-L1 axis binding antagonist (e.g., as described in Section III below).

[0235] For example, provided herein are methods for treating cancer including the treatment of cancer with an anti-cancer therapy comprising a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). A method for identifying an individual having cancer (e.g., renal cancer (e.g., RCC)) who may benefit, the method comprising determining whether the individual has a sarcomatoid cancer, wherein the presence of a sarcomatoid cancer identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0236] In another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising (a) determining whether the individual has sarcomatoid cancer, wherein the presence of sarcomatoid cancer indicates treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab) or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis-binding antagonist (e.g., ... and (b) identifying the individual as one who would benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the presence of sarcomatoid cancer. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0237] The benefit may relate, for example, to improved progression-free survival (PFS), overall survival (OS), overall response rate (ORR), complete response (CR) rate, or disease progression-free rate (DFR). In some embodiments, the benefit relates to improved PFS. In some cases, the benefit relates to improved OS. In some cases, the benefit relates to improved ORR. In some cases, the benefit relates to improved CR rate. In some cases, the benefit relates to improved DFR. In some cases, the DFR is determined as the time from the start of treatment to the individual's first increase of 2 or more points above baseline on the MD Anderson Symptom Assessment Index (MDASI) disability scale.

[0238] For example, provided herein are cancer cells that may benefit from treatment with anti-cancer therapies including a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)),

[0010] In some embodiments, the present invention relates to a method for identifying an individual having a cancer (e.g., a renal cancer (e.g., RCC)), the method comprising determining whether the individual has a sarcomatoid cancer, wherein the presence of a sarcomatoid cancer identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, wherein the benefit is related to improved PFS. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0239] In another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising (a) determining whether the individual has sarcomatoid cancer, wherein the presence of sarcomatoid cancer indicates treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab) or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis-binding antagonist (e.g., ... and (b) identifying the individual as one who would benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the presence of sarcomatoid cancer. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0240] In another example, provided herein is a cancer treatment for a patient that may benefit from treatment with an anti-cancer therapy including a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). and a method for identifying an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising determining whether the individual has sarcomatoid cancer, wherein the presence of sarcomatoid cancer identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the benefit being related to improved OS. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0241] In yet another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising (a) determining whether the individual has a sarcomatoid cancer, wherein the presence of a sarcomatoid cancer indicates a potential benefit from treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., For example, the method includes (a) identifying the individual as one who would benefit from treatment with an anti-cancer therapy comprising an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody), and determining that benefit is in terms of improved OS, and (b) selecting an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist based on the presence of sarcomatoid cancer. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist.

[0242] In further examples, provided herein are cancer patients that may benefit from treatment with an anti-cancer therapy including a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). and a method for identifying an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising determining whether the individual has sarcomatoid cancer, wherein the presence of sarcomatoid cancer identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, wherein the benefit is associated with improved ORR. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0243] In yet another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising (a) determining whether the individual has a sarcomatoid cancer, wherein the presence of a sarcomatoid cancer indicates a potential benefit from treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., For example, the method includes (a) identifying the individual as one who would benefit from treatment with an anti-cancer therapy comprising an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody), and determining whether the benefit is in terms of improved ORR; and (b) selecting an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the presence of sarcomatoid cancer. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0244] In yet another example, provided herein is a cancer therapy that may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

[0010] A method for identifying an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising determining whether the individual has sarcomatoid carcinoma, wherein the presence of sarcomatoid carcinoma identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, wherein the benefit is related to an improved CR rate. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0245] In another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising (a) determining whether the individual has sarcomatoid cancer, wherein the presence of sarcomatoid cancer indicates a potential benefit from treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., and (b) identifying the individual as one who would benefit from treatment with an anti-cancer therapy comprising an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody), where the benefit is in terms of an improved CR rate, and (b) selecting an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist based on the presence of sarcomatoid cancer. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist.

[0246] In yet another example, provided herein is a cancer therapy that may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

[0010] A method for identifying an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising determining whether the individual has sarcomatoid carcinoma, wherein the presence of sarcomatoid carcinoma identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, wherein the benefit is associated with an improved DFR. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0247] In another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., renal cancer (e.g., RCC)), the method comprising (a) determining whether the individual has sarcomatoid cancer, wherein the presence of sarcomatoid cancer indicates a potential benefit from treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., and (b) identifying the individual as one who would benefit from treatment with an anti-cancer therapy comprising an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody), and determining whether the benefit is in terms of improved DFR, and (b) selecting an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist based on the presence of sarcomatoid cancer. In some embodiments, the method further comprises administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist.

[0248] The presence of sarcomatoid cancer (e.g., sarcomatoid renal cancer (e.g., sarcomatoid RCC)) can be determined using any suitable approach. See, e.g., El Mouallem et al., Urol. Oncol. 36:265-271, 2018. For example, in some embodiments, the presence of sarcomatoid cancer (e.g., sarcomatoid renal cancer (e.g., sarcomatoid RCC)) is assessed by histological analysis of a sample obtained from an individual. In some embodiments, the renal cancer is sarcomatoid if a tumor sample from the individual contains foci(s) of any component high-grade malignant spindle cells relative to the overall tumor area. In some embodiments, the spindle cells exhibit moderate to marked atypia and / or resemble any form of sarcoma. In some embodiments, the spindle cells show evidence of epithelial differentiation, as assessed by immunohistological positivity for keratin or epithelial membrane antigen (EMA). In some embodiments, the renal cancer is renal cell carcinoma, and the tumor sample has epithelial differentiation with concurrent areas of renal cell carcinoma.

[0249] In any of the above methods, the method may further comprise determining the individual's MSKCC risk score. In other embodiments, the individual's MSKCC risk score has been previously determined. In any of the above methods, the individual may have a low or intermediate MSKCC risk score.

[0250] In another example, provided herein is a method of identifying an individual having cancer (e.g., renal cancer (e.g., RCC)) who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))) and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining the individual's MSKCC risk score, wherein a low or intermediate MSKCC risk score identifies the individual as likely to benefit from anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0251] In yet another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., kidney cancer (e.g., RCC)), the method comprising: (a) determining the individual's MSKCC risk score, wherein a low or moderate MSKCC risk score indicates a risk factor for treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab) or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)). and (b) identifying, assessing the individual as likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), and (b) selecting an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the individual having a low or intermediate MSKCC risk score.

[0252] The benefit may relate, for example, to improved progression-free survival (PFS), overall survival (OS), overall response rate (ORR), complete response (CR) rate, or disease progression-free rate (DFR). In some embodiments, the benefit relates to improved PFS. In some cases, the benefit relates to improved OS. In some cases, the benefit relates to improved ORR. In some cases, the benefit relates to improved CR rate. In some cases, the benefit relates to improved DFR. In some cases, the DFR is determined as the time from the start of treatment to the individual's first increase of 2 or more points above baseline on the MD Anderson Symptom Assessment Index (MDASI) disability scale.

[0253] For example, provided herein are therapeutic agents that combine a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))) and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab), or a PD-1 binding antagonist (e.g., 1. A method for identifying an individual having cancer (e.g., kidney cancer (e.g., RCC)) who may benefit from treatment with an anti-cancer therapy comprising an anti-PD-1 antibody), the method comprising determining the individual's MSKCC risk score, wherein a low or intermediate MSKCC risk score identifies the individual as likely to benefit from an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, wherein the benefit is associated with improved PFS.

[0254] In another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., kidney cancer (e.g., RCC)), the method comprising (a) determining the individual's MSKCC risk score, wherein a low or moderate MSKCC risk score indicates a risk of progression to a cancer that is associated with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab) or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis inhibitor. and (b) identifying an individual as likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), and determining whether the benefit is in terms of improved PFS; and (b) selecting an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist based on the individual having a low or intermediate MSKCC risk score.

[0255] In another example, provided herein is a combination of a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))) and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab), or a PD-1 binding antagonist (e.g., 1. A method for identifying an individual having cancer (e.g., kidney cancer (e.g., RCC)) who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the method comprising determining the individual's MSKCC risk score, wherein a low or intermediate MSKCC risk score identifies the individual as likely to benefit from an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the benefit being associated with improved OS.

[0256] In yet another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., kidney cancer (e.g., RCC)), the method comprising (a) determining the individual's MSKCC risk score, wherein a low or moderate MSKCC risk score indicates a potential treatment option for a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L and (b) identifying an individual as likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist (e.g., a PD-L1-binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1-binding antagonist (e.g., an anti-PD-1 antibody)), and determining that benefit is in terms of improved OS; and (b) selecting an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the individual having a low or intermediate MSKCC risk score.

[0257] In further examples, provided herein are therapeutic agents that combine a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))) and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab), or a PD-1 binding antagonist (e.g., 1. A method for identifying an individual having cancer (e.g., kidney cancer (e.g., RCC)) who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the method comprising determining an MSKCC risk score for the individual, wherein a low or intermediate MSKCC risk score identifies the individual as likely to benefit from an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the benefit being associated with an improved ORR.

[0258] In yet another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., kidney cancer (e.g., RCC)), the method comprising (a) determining the individual's MSKCC risk score, wherein a low or moderate MSKCC risk score indicates a potential treatment option for a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L and (b) identifying an individual as likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist (e.g., a PD-L1-binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1-binding antagonist (e.g., an anti-PD-1 antibody)), and determining whether the benefit is in terms of improved ORR; and (b) selecting an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the individual having a low or intermediate MSKCC risk score.

[0259] In yet another example, provided herein is a combination of a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))) and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab), or a PD-1 binding antagonist (e.g., a PD-L1 antibody, e.g., atezolizumab). and a VEGF antagonist and a PD-L1 axis-binding antagonist), the method comprising determining an MSKCC risk score for the individual, wherein a low or intermediate MSKCC risk score identifies the individual as likely to benefit from anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, wherein the benefit is associated with an improved CR rate.

[0260] In yet another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., kidney cancer (e.g., RCC)), the method comprising (a) determining the individual's MSKCC risk score, wherein a low or moderate MSKCC risk score indicates a potential treatment option for a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L and (b) identifying an individual as likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist (e.g., a PD-L1-binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1-binding antagonist (e.g., an anti-PD-1 antibody)), and determining whether said benefit is in terms of improved CR rate; and (b) selecting an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the individual having a low or intermediate MSKCC risk score.

[0261] In another example, provided herein is a combination of a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))) and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab), or a PD-1 binding antagonist (e.g., , anti-PD-1 antibody)) the method of identifying an individual having cancer (e.g., kidney cancer (e.g., RCC)) who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the method comprising determining the individual's MSKCC risk score, wherein a low or intermediate MSKCC risk score identifies the individual as likely to benefit from an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist, the benefit being associated with an improved DFR.

[0262] In yet another example, provided herein is a method for selecting a treatment for an individual having cancer (e.g., kidney cancer (e.g., RCC)), the method comprising: (a) determining the individual's MSKCC risk score; and determining whether a low or moderate MSKCC risk score is a good indication for treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab) or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 inhibitor. (b) identifying the individual as likely to benefit from an anti-cancer treatment comprising an axis-binding antagonist (e.g., a PD-L1-binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab, or a PD-1-binding antagonist (e.g., an anti-PD-1 antibody)), and determining whether the benefit is in terms of improved DFR; and (b) selecting an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis-binding antagonist based on the individual having a low or intermediate MSKCC risk score.

[0263] In any of the foregoing methods, an individual may have a high MSKCC risk score if the individual has three or more (e.g., three, four, or all five) of the following characteristics: (i) time from nephrectomy to systemic treatment less than one year, absence of nephrectomy, or initial diagnosis of metastatic disease; (ii) hemoglobin levels below the lower limit of normal (LLN) (in some cases, the normal range for hemoglobin is 13.5-17.5 g / dL for men and 12-15.5 g / dL for women). (iii) a corrected serum calcium level greater than 10 mg / dL (optionally, the corrected serum calcium level is serum calcium level (mg / dL) + 0.8(4 - serum albumin (g / dL))); (iv) a serum lactate dehydrogenase (LDH) level greater than 1.5 times the upper limit of normal (ULN) (optionally, the ULN is 140 U / L); and / or (v) a Karnofsky Performance Status (KPS) score less than 80. In some embodiments, the individual has three of the foregoing characteristics. In other embodiments, the individual has four of the foregoing characteristics. In yet other embodiments, the individual has all five of the foregoing characteristics.

[0264] In any of the foregoing methods, an individual may have a moderate MSKCC risk score if the individual has one or two of the following characteristics: (i) time from nephrectomy to systemic treatment less than one year, absence of nephrectomy, or initial diagnosis of metastatic disease; (ii) a hemoglobin level below the LLN (optionally, the normal range for hemoglobin is 13.5-17.5 g / dL for men and 12-15.5 g / dL for women); (iii) a corrected serum calcium level greater than 10 mg / dL (optionally, the corrected serum calcium level is serum calcium level (mg / dL) + 0.8 (4 - serum albumin (g / dL))); (iv) a serum LDH level greater than 1.5 times the ULN (optionally, the ULN is 140 U / L); and / or (v) a KPS score less than 80. In some embodiments, the individual has one of the foregoing characteristics. In other embodiments, the individual has two of the foregoing characteristics.

[0265] In any of the foregoing methods, the individual can have a sarcomatoid cancer (eg, a sarcomatoid renal cancer (eg, a sarcomatoid RCC)).

[0266] In some embodiments of any of the aforementioned methods, the method further comprises determining the expression level of one or more of the genes listed in Table 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, or 37). In other embodiments, the expression level of one or more of the genes listed in Table 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, or 37) has been previously determined. Table 1. Exemplary biomarkers TIFF2025041681000001.tif173170

[0267] For example, in some embodiments, the method further comprises determining 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, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33) of the following genes in a sample from the individual: CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2; VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34; or IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2. In other embodiments, the expression level of one or more of the following genes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33) in a sample from the individual has been previously determined: CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2; VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34; or IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2.

[0268] In some embodiments of any of the foregoing methods, (i) an 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 20) of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample that is equal to or greater than a reference expression level of the one or more genes. or (ii) an expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13) of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34, or IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample that is less than the reference expression level of the one or more genes identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis-binding antagonist.

[0269] In any of the foregoing methods, the method may include determining 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 20) of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2. In some embodiments, the method comprises determining the expression level of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or all twenty of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2.

[0270] For example, any of the foregoing methods may include determining the expression level of one or more (e.g., 1, 2, 3, 4, or 5) of CD8A, EOMES, PRF1, IFNG, or PD-L1. In some embodiments, the method includes determining the expression level of at least two, at least three, at least four, or all five of CD8A, EOMES, PRF1, IFNG, and PD-L1. In some embodiments, the method includes determining the expression level of two of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 2). In some embodiments, the method includes determining the expression level of three of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 3). In some embodiments, the method includes determining the expression level of four of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 4). In some embodiments, the method comprises determining the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1. [Table 2] Combinations of two of the following genes: CD8A, EOMES, PRF1, IFNG, and PD-L1 TIFF2025041681000002.tif50170[Table 3] Combinations of three genes among CD8A, EOMES, PRF1, IFNG, and PD-L1 TIFF2025041681000003.tif63170[Table 4] Combinations of four genes: CD8A, EOMES, PRF1, IFNG, and PD-L1 TIFF2025041681000004.tif29170

[0271] In some embodiments, any of the foregoing methods may include determining the expression level of PD-L1 and one or more additional genes, wherein the one or more additional genes are other than PD-L1. For example, in some embodiments, a method may include determining the expression level of PD-L1 and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36) additional genes selected from the group consisting of: CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8 In some embodiments, the method comprises determining the expression level of PD-L1 and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19) additional genes selected from the group consisting of: PSMB9, TAP1, TAP2, VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, CD34, IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9. In other embodiments, the method comprises determining the expression levels of PD-L1 and one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34. In other embodiments, the method comprises determining the expression levels of PD-L1 and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9.

[0272] Any of the foregoing methods may include determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34. In some embodiments, the method includes determining the expression level of at least two, at least three, at least four, at least five, at least six, or all seven of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34. For example, in some embodiments, the method includes determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, or 6) of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, or CD34. In some embodiments, the method includes determining the expression level of at least two, at least three, at least four, at least five, or all six of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34. In some embodiments, the method comprises determining the expression levels of two of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 5). In some embodiments, the method comprises determining the expression levels of three of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 6). In some embodiments, the method comprises determining the expression levels of four of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 7). In some embodiments, the method comprises determining the expression levels of five of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 8). In some embodiments, the method comprises determining the expression levels of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34. Table 5. Combinations of two genes from VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 TIFF2025041681000005.tif71170[Table 6] Combinations of three genes among VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 TIFF2025041681000006.tif94170 [Table 7] Combinations of four genes among VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 TIFF2025041681000007.tif74170 [Table 8] Combination of five genes among VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 TIFF2025041681000008.tif31170

[0273] Any of the foregoing methods can include determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9. In some embodiments, the method includes determining the expression level of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all ten of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9. In some embodiments, the method comprises determining the expression levels of two of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 9). In some embodiments, the method comprises determining the expression levels of three of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 10). In some embodiments, the method comprises determining the expression levels of four of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 11). In some embodiments, the method comprises determining the expression levels of five of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 12). In some embodiments, the method comprises determining the expression levels of six of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 13).In some embodiments, the method comprises determining the expression levels of seven of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 14). In some embodiments, the method comprises determining the expression levels of eight of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 15). In some embodiments, the method comprises determining the expression levels of nine of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 16). In some embodiments, the method comprises determining the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9. [Table 9] Combinations of two genes from IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 TIFF2025041681000009.tif204170[Table 10] Combinations of three genes from IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 TIFF2025041681000010.tif255170TIFF2025041681000011.tif255170TIFF2025041681000012.tif16170[Table 11] Combinations of four genes from IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 TIFF2025041681000013.tif227170TIFF2025041681000014.tif255170TIFF2025041681000015.tif255170TIFF2025041681000016.tif180170[Table 12] Combinations of five genes among IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 TIFF2025041681000017.tif255170TIFF2025041681000018.tif255170TIFF2025041681000019.tif255170TIFF2025041681000020.tif255170TIFF2025041681000021.tif76170[Table 13] Combination of six genes among IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 TIFF2025041681000022.tif162170TIFF2025041681000023.tif255170TIFF2025041681000024.tif255170TIFF2025041681000025.tif237170[Table 14] Combination of seven genes among IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 TIFF2025041681000026.tif254170TIFF2025041681000027.tif255170TIFF2025041681000028.tif255170TIFF2025041681000029.tif255170[Table 15] Combination of eight genes among IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 TIFF2025041681000030.tif239170TIFF2025041681000031.tif129170[Table 16] Combinations of nine genes among IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 TIFF2025041681000032.tif88170

[0274] In any of the foregoing methods, the method may include determining 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 20) of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2, and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9. For example, in some embodiments, the method includes detecting at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least one of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, and TAP2. determining the expression levels of at least 3, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or all 20 of the following: IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9.

[0275] For example, any of the foregoing methods may include determining the expression levels of one or more (e.g., 1, 2, 3, 4, or 5) of CD8A, EOMES, PRF1, IFNG, or PD-L1, and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9. In some embodiments, the method comprises determining the expression levels of at least two, at least three, at least four, or all five of CD8A, EOMES, PRF1, IFNG, and PD-L1, and at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all ten of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9. In some embodiments, the method comprises determining the expression levels of any one of the combinations set forth in Tables 2-4 and any one of the combinations set forth in Tables 9-16. For example, in some embodiments, the method comprises determining the expression levels of two of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 2), and two of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 9). In some embodiments, the method comprises determining the expression levels of three of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 3), and three of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 10).In some embodiments, the method comprises determining the expression levels of four of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 4), and four of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 11). In some embodiments, the method comprises determining the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and five of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 12). In some embodiments, the method comprises determining the expression levels of six of CD8A, EOMES, PRF1, IFNG, and PD-L1, and IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 13). In some embodiments, the method comprises determining the expression levels of seven of CD8A, EOMES, PRF1, IFNG, and PD-L1, and IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 14). In some embodiments, the method comprises determining the expression levels of eight of CD8A, EOMES, PRF1, IFNG, and PD-L1, and IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 15). In some embodiments, the method comprises determining the expression levels of nine of CD8A, EOMES, PRF1, IFNG, and PD-L1, and IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 16).In some embodiments, the method comprises determining the expression levels of CD8A, EOMES, PRF1, IFNG, PD-L1, IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9.

[0276] In other embodiments, in any of the foregoing methods, the method may include determining 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 20) of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2, and one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34. For example, in some embodiments, a method comprises determining the expression levels of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or all 20 of: CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, and TAP2; and at least two, at least three, at least four, at least five, at least six, or all seven of: VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34.

[0277] For example, any of the aforementioned methods may include determining the expression levels of one or more (e.g., 1, 2, 3, 4, or 5) of CD8A, EOMES, PRF1, IFNG, or PD-L1, and one or more (e.g., 1, 2, 3, 4, 5, or 6) of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, or CD34. In some embodiments, the method includes determining the expression levels of at least two, at least three, at least four, or all five of CD8A, EOMES, PRF1, IFNG, and PD-L1, and at least one, at least two, at least three, at least four, at least five, or all six of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34. In some embodiments, the method includes determining the expression levels of any one of the combinations listed in Tables 2-4 and any one of the combinations listed in Tables 5-8. For example, in some embodiments, the method comprises determining the expression levels of two of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 2), and two of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 5). In some embodiments, the method comprises determining the expression levels of three of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 3), and three of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 6). In some embodiments, the method includes determining the expression levels of four of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 4), and four of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 7).In some embodiments, the method comprises determining the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and five of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 8). In some embodiments, the method comprises determining the expression levels of CD8A, EOMES, PRF1, IFNG, PD-L1, VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34.

[0278] In further embodiments, in any of the foregoing methods, the method may include determining the expression levels of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9, and one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34. For example, in some embodiments, the methods include determining the expression levels of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all ten of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9, and at least two, at least three, at least four, at least five, at least six, or all seven of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34.

[0279] For example, any of the foregoing methods may include determining the expression levels of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9, and one or more (e.g., 1, 2, 3, 4, 5, or 6) of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, or CD34. In some embodiments, the method comprises determining the expression levels of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all ten of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9, and at least two, at least three, at least four, at least five, or all six of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34. In some embodiments, the method comprises determining the expression levels of any one of the combinations set forth in Tables 9-16 and any one of the combinations set forth in Tables 5-8. For example, in some embodiments, the method includes determining the expression levels of two of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 9), and two of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 5). In some embodiments, the method includes determining the expression levels of three of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 10), and three of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 6).In some embodiments, the method comprises determining the expression levels of four of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 11), and four of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 7). In some embodiments, the method comprises determining the expression levels of five of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 12), and five of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 8). In some embodiments, the method comprises determining the expression levels of six of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 13), as well as VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34. In some embodiments, the method comprises determining the expression levels of seven of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 14), as well as VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34. In some embodiments, the method includes determining the expression levels of eight of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 15), as well as VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34.In some embodiments, the method comprises determining the expression levels of nine of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 16), as well as VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34. In some embodiments, the method comprises determining the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, S100A9, VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34.

[0280] In some embodiments of any of the aforementioned methods, 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 20) of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample is equal to or greater than a reference expression level of the one or more genes, and the method further comprises administering to the individual an effective amount of an anti-cancer therapy. For example, in some embodiments, the expression level of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or all twenty of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 is equal to or greater than the reference expression level of the one or more genes. In some cases, the expression level of one or more (e.g., 1, 2, 3, 4, or 5) of CD8A, EOMES, PRF1, IFNG, or PD-L1 in the sample is equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression level of one or more of the exemplary combinations shown in Tables 2-4 in the sample is equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 in the sample are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1.

[0281] In some embodiments of any of the foregoing methods, the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 in the sample is equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression level of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all ten of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 in the sample is equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression level of one or more of the exemplary combinations shown in Tables 9-16 in the sample is equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 in the sample are equal to or greater than the reference expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9.

[0282] In some embodiments of any of the aforementioned methods, one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 19, or 20) is equal to or greater than the baseline expression level of the one or more genes, and the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 is equal to or greater than the baseline expression level of the one or more genes, and the method further comprises administering to the individual an effective amount of an anti-cancer therapy. For example, in some embodiments, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least 12, at least 13, at least 14, at least 15, or at least a few of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, and TAP2 are identified. The expression levels of at least 16, at least 17, at least 18, at least 19, or all 20 genes are equal to or greater than the reference expression level of the one or more genes, and the expression levels of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all 10 of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 are equal to or greater than the reference expression level of the one or more genes.In some embodiments, the presence of osteomyelitis in the tumor is identified by an expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 that is equal to or greater than the reference expression level of the one or more genes.

[0283] For example, in some embodiments, the expression level of one or more (e.g., 1, 2, 3, 4, or 5) of CD8A, EOMES, PRF1, IFNG, or PD-L1 is equal to or greater than the reference expression level of the one or more genes, and the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 is equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression levels of at least two, at least three, at least four, or all five of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression level of the one or more genes, and the expression levels of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all ten of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 are equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression level of any one of the combinations shown in Tables 2-4 is equal to or greater than the reference expression level of the one or more genes, and the expression level of any one of the combinations shown in Tables 9-16 is equal to or greater than the reference expression level of the one or more genes. For example, in some embodiments, the expression levels of two of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 2) are equal to or greater than the reference expression level of the one or more genes, and the expression levels of two of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 9) are equal to or greater than the reference expression level of the one or more genes.In some embodiments, the expression levels of three of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 3) are equal to or greater than the reference expression level of the one or more genes, and the expression levels of three of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 10) are equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression levels of four of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 4) are equal to or greater than the reference expression level of the one or more genes, and the expression levels of four of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 11) are equal to or greater than the reference expression level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of five of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 12) are equal to or greater than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of six of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 13) are equal to or greater than the reference level of the one or more genes.In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of seven of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 14) are equal to or greater than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of eight of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 15) are equal to or greater than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of nine of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 16) are equal to or greater than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, PD-L1, IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, PD-L1, IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9.In some embodiments, the presence of osteomyelitis in the tumor is identified by an expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 that is equal to or greater than the reference expression level of the one or more genes. In some embodiments, 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 20) of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample are equal to or greater than the reference expression level of the one or more genes. and an expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 that is equal to or greater than the baseline expression level of the one or more genes indicates that the individual is unlikely to benefit (e.g., is resistant) to a PD-L1 axis-binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab) monotherapy.

[0284] In other embodiments of any of the foregoing methods, 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 20) of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample indicates a level of expression of the one or more genes. or greater than the baseline expression level of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9, and the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 is less than the baseline expression level of said one or more genes, and the method further comprises administering to the individual an effective amount of a PD-L1 axis-binding antagonist (e.g., an anti-PD-L1 antibody (e.g., atezolizumab) or an anti-PD-1 antibody) monotherapy. For example, in some embodiments, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least 12, at least 13, at least 14, at least 15, or at least a few of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, and TAP2 are identified. The expression levels of at least 16, at least 17, at least 18, at least 19, or all 20 genes are equal to or greater than the reference expression level of the one or more genes, and the expression levels of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all 10 of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 are less than the reference expression level of the one or more genes.

[0285] For example, in some embodiments, the expression level of one or more (e.g., 1, 2, 3, 4, or 5) of CD8A, EOMES, PRF1, IFNG, or PD-L1 is equal to or greater than the reference expression level of the one or more genes, and the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 is less than the reference expression level of the one or more genes. In some embodiments, the expression levels of at least two, at least three, at least four, or all five of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression level of the one or more genes, and the expression levels of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all ten of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 are less than the reference expression level of the one or more genes. In some embodiments, the expression level of any one of the combinations shown in Tables 2-4 is equal to or greater than the reference expression level of the one or more genes, and the expression level of any one of the combinations shown in Tables 9-16 is less than the reference expression level of the one or more genes. For example, in some embodiments, the expression levels of two of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 2) are equal to or greater than the reference expression level of the one or more genes, and the expression levels of two of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 9) are less than the reference expression level of the one or more genes.In some embodiments, the expression levels of three of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 3) are equal to or greater than the reference expression level of the one or more genes, and the expression levels of three of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 10) are less than the reference expression level of the one or more genes. In some embodiments, the expression levels of four of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 4) are equal to or greater than the reference expression level of the one or more genes, and the expression levels of four of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 11) are less than the reference expression level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of five of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 12) are less than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of six of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 13) are less than the reference level of the one or more genes.In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of seven of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 14) are less than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of eight of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 15) are less than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of nine of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 (e.g., any of the exemplary combinations shown in Table 16) are less than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 are less than the reference expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9.

[0286] In some embodiments of any of the foregoing methods, the expression level of PD-L1 in the sample is equal to or greater than a reference expression level of PD-L1, and 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, or 19) additional genes selected from the group consisting of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample is equal to or greater than a reference expression level of the one or more additional genes.

[0287] In some embodiments of any of the foregoing methods, the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 in the sample is less than the reference level for the one or more genes, and the method further comprises administering an effective amount of an anti-cancer treatment to the individual. For example, in some embodiments, the expression level of at least one, at least two, at least three, at least four, at least five, at least six, or all seven of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 in the sample is less than the reference level for the one or more genes. In some embodiments, the expression level of one or more of the exemplary combinations shown in Tables 5-8 in the sample is less than the reference expression level for the one or more genes. In some embodiments, the expression level of one or more of VEGFA, KDR, ESM1, PECAM1, ANGPTL4 or CD34 in sample is less than the reference level of said one or more genes.For example, in some embodiments, the expression level of VEGFA, KDR, ESM1, PECAM1, ANGPTL4 and CD34 in sample is less than the reference level of VEGFA, KDR, ESM1, PECAM1, ANGPTL4 and CD34.

[0288] In other embodiments, in any of the foregoing methods, one or more of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) is equal to or greater than the reference level for the one or more genes, and the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 is less than the reference level for the one or more genes, and the method further comprises administering to the individual an effective amount of an anti-cancer therapy. For example, in some embodiments, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, or at least twelve of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, and TAP2 are selected from the group consisting of: , at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or all 20 of the following genes are at or above the reference level for the one or more genes: VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34; and the expression levels of at least two, at least three, at least four, at least five, at least six, or all seven of the following genes are below the reference level for the one or more genes:

[0289] For example, in some embodiments, the expression level of one or more (e.g., 1, 2, 3, 4, or 5) of CD8A, EOMES, PRF1, IFNG, or PD-L1 is equal to or greater than the reference level for the one or more genes, and the expression level of one or more (e.g., 1, 2, 3, 4, 5, or 6) of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, or CD34 is less than the reference level for the one or more genes. In some embodiments, the expression levels of at least two, at least three, at least four, or all five of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference level for the one or more genes, and the expression levels of at least one, at least two, at least three, at least four, at least five, or all six of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 are less than the reference level for the one or more genes. In some embodiments, the expression level of any one of the combinations shown in Tables 2-4 is equal to or greater than the reference level of the one or more genes, and the expression level of any one of the combinations shown in Tables 5-8 is less than the reference level of the one or more genes. For example, in some embodiments, the expression level of two of CD8A, EOMES, PRFl, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 2) is equal to or greater than the reference level of the one or more genes, and the expression level of two of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 5) is less than the reference level of the one or more genes. In some embodiments, the expression levels of three of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 3) are equal to or greater than the reference level of the one or more genes, and the expression levels of three of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 6) are less than the reference level of the one or more genes.In some embodiments, the expression levels of four of CD8A, EOMES, PRF1, IFNG, and PD-L1 (e.g., any of the exemplary combinations shown in Table 4) are equal to or greater than the reference level of the one or more genes, and the expression levels of four of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 7) are less than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference level of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of five of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 (e.g., any of the exemplary combinations shown in Table 8) are less than the reference level of the one or more genes. In some embodiments, the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 are equal to or greater than the reference levels of CD8A, EOMES, PRF1, IFNG, and PD-L1, and the expression levels of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 are less than the reference levels of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34.

[0290] In some embodiments of any of the aforementioned methods, the expression level of one or more (e.g., 1, 2, 3, 4, 5, or 6) of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 in the sample is less than the reference level of the one or more genes, and the method further comprises administering an effective amount of an anti-cancer treatment to the individual. In some embodiments, the expression level of at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all ten of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9 in the sample is less than the reference level of the one or more genes. For example, in some embodiments, the expression levels of one or more of the exemplary combinations shown in Tables 9-16 in a sample are less than the reference expression levels of the one or more genes. In some embodiments, the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9 in a sample are less than the reference levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and S100A9.

[0291] In other embodiments of any of the foregoing methods, the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 in the sample is equal to or greater than a reference level for the one or more genes, and the method further comprises administering to the individual an effective amount of an angiogenesis inhibitor (e.g., a VEGF antagonist (e.g., a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))). In some embodiments, the expression level of at least one, at least two, at least three, at least four, at least five, at least six, or all seven of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 is equal to or greater than a reference level for the one or more genes. In some embodiments, the expression levels of one or more (e.g., 1, 2, 3, 4, 5, or 6) of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, or CD34 in the sample are equal to or greater than the reference level for the one or more genes. In some embodiments, the expression levels of one or more of the exemplary combinations shown in Tables 5-8 in the sample are equal to or greater than the reference expression level for the one or more genes. In some embodiments, the expression levels of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 in the sample are equal to or greater than the reference levels for VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34.

[0292] In certain embodiments of any of the foregoing methods, the reference level is a reference population (e.g., a population of individuals with cancer (e.g., kidney cancer (e.g., RCC))) 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, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, or 37) genes (e.g., CD8A, EOMES, GZMA, GZ, In certain embodiments, the reference level is the expression level of one or more genes in a reference population, such as a reference group of individuals with cancer. In other embodiments, the reference level may be the top 40%, top 30%, top 20%, top 10%, top 5%, or top 1% of the expression levels in the reference population. In certain embodiments, reference level is the predetermined expression level of one or more genes.In some embodiments, reference level is the median Z-score of the normalized expression level of one or more genes.In some embodiments, reference level is the expression level of one or more genes in the biological sample obtained from patient at a previous time point, wherein the previous time point is after administering anti-cancer treatment.In some embodiments of any of the foregoing methods, the reference level is the expression level of one or more genes (e.g., CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2; VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34; or IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, or S100A9) in a biological sample from the patient obtained prior to administration of the anti-cancer treatment (e.g., minutes, hours, days, weeks (e.g., 1, 2, 3, 4, 5, 6, or 7 weeks), months, or years). In other embodiments, the reference level is the expression level of one or more genes in a biological sample obtained from the patient at a later time point (e.g., minutes, hours, days, weeks, months, or years after administration of the anti-cancer treatment).

[0293] The presence and / or expression level of any of the above biomarkers can be assessed qualitatively and / or quantitatively based on any suitable criteria known in the art, including, but not limited to, DNA, mRNA, cDNA, protein, protein fragment, and / or gene copy number. Methodologies for measuring such biomarkers are known in the art and understood by those skilled in the art. Such methodologies include, but are not limited to, 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 (ISH), fluorescent in situ hybridization (FISH), Southern analysis, Northern analysis, whole genome sequencing, polymerase chain reaction (PCR), such as quantitative real-time PCR (qRT-PCR) and other amplification-based detection methods, such as branched DNA, SISBA, and TMA, RNA-Seq, microarray analysis, gene expression profiling, whole genome sequencing (WGS), and / or serial analysis of gene expression ("SAGE"), and any one of a wide variety of assays that can be performed by protein, gene, and / or tissue array analysis. Exemplary protocols for assessing the status of genes and gene products are found, for example, in Ausubel et al. (Current Protocols In Molecular Biology, 1995), Units 2 (Northern blotting), 4 (Southern blotting), 15 (immunoblotting), and 18 (PCR analysis). Multiplexed immunoassays, such as those available from Rules Based Medicine or Meso Scale Discovery ("MSD"), can also be used.

[0294] In some embodiments of any of the foregoing methods, the expression level of the biomarker may be a nucleic acid expression level (e.g., a DNA expression level) or an RNA expression level (e.g., an mRNA expression level). Any suitable method of determining nucleic acid expression levels may be used. In some embodiments, the nucleic acid expression levels are determined using RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

[0295] Methods for assessing mRNA in cells are well known and include, for example, serial analysis of gene expression (SAGE), whole genome sequencing (WGS), hybridization assays using complementary DNA probes (such as in situ hybridization, Northern blots, and related techniques using labeled riboprobes specific for one or more genes), and various nucleic acid amplification assays (such as RT-PCR (e.g., qRT-PCR) using complementary primers specific for one or more genes, and other amplification-based detection methods such as branched DNA, SISBA, and TMA). Additionally, such methods may include one or more steps that allow for the determination of the level of a target mRNA in a biological sample (e.g., by simultaneously testing the level of a control mRNA sequence of a "housekeeping" gene, such as an actin family member). In some cases, the sequence of the amplified target cDNA may be determined. Some methods include protocols for testing or detecting mRNAs, such as target mRNAs, in tissue or cell samples using microarray technology. Using nucleic acid microarray, test and control mRNA samples from test and control tissue samples are reverse transcribed and labeled to generate cDNA probes.The probes are then hybridized to the array of nucleic acids fixed on solid support.The array is configured so that the sequence and position of each array member can be identified.For example, a selection of genes whose expression correlates with the increase or decrease of clinical benefit of treatments that include VEGF antagonists and PD-L1 axis binding antagonists can be arrayed on solid support.The hybridization of a specific array member with a labeled probe indicates that the sample from which the probe is derived expresses that gene.

[0296] In other embodiments of any of the foregoing methods, the expression level of the biomarker may be a protein expression level. In certain embodiments, the method comprises contacting the sample with an antibody that specifically binds to a biomarker described herein under conditions that allow binding of the biomarker, and detecting whether a complex is formed between the antibody and the biomarker. Such methods may be in vitro or in vivo methods. In some cases, the antibodies are used to select patients eligible for treatment with a VEGF antagonist (e.g., an anti-VEGF antibody (e.g., bevacizumab), or a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib))), and a PD-L1 axis-binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atezolizumab (MPDL3280A)), or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), e.g., in conjunction with a biomarker for individual selection. In other cases, the antibody is used in conjunction with an angiogenesis inhibitor (e.g., a VEGF antagonist (e.g., a VEGFR inhibitor (e.g., a multi-targeted tyrosine kinase inhibitor (e.g., sunitinib, axitinib, pazopanib, or cabozantinib)))), e.g., a biomarker for the selection of individuals to select patients eligible for treatment. Any method of measuring protein expression levels known in the art or provided herein may be used. For example, in some embodiments, the protein expression level of the biomarker is determined using a method selected from the group consisting of flow cytometry (e.g., fluorescence-activated cell sorting (FACS™)), Western blot, enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, immunohistochemistry (IHC), immunofluorescence, radioimmunoassay, dot blotting, immunodetection methods, HPLC, surface plasmon resonance, optical spectroscopy, mass spectrometry, and HPLC. In some embodiments, the protein expression level of the biomarker is determined in tumor-infiltrating immune cells. In some embodiments, the protein expression level of the biomarker is determined in tumor cells.In some embodiments, the protein expression level of the biomarker is determined in tumor-infiltrating immune cells and / or tumor cells. In some embodiments, the protein expression level of the biomarker is determined in peripheral blood mononuclear cells (PBMCs).

[0297] In certain embodiments, the presence and / or expression level / amount of biomarker proteins in a sample are tested using IHC and staining protocols. IHC staining of tissue sections has been shown to be a reliable method for determining or detecting the presence of proteins in a sample. In some embodiments of any of the methods, assays, and / or kits, the biomarker is one or more of the protein expression products of the following genes: CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, TAP2, VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, CD34, IL6, CXCL1, CXCL2, CXCL3, CXCL8, PTGS2, CXCR1, CXCR2, S100A8, and / or S100A9. In one embodiment, the expression level of a biomarker is determined using a method comprising: (a) performing an IHC analysis of a sample (such as a tumor sample obtained from a patient) with an antibody; and (b) determining the expression level of the biomarker in the sample. In some embodiments, the IHC staining intensity is determined relative to a reference. In some embodiments, the reference is a reference value. In some embodiments, the reference is a reference sample (e.g., a control cell line staining sample, a tissue sample from a non-cancerous patient, or a tumor sample determined to be negative for the biomarker of interest).

[0298] IHC may be performed in combination with additional techniques such as morphological staining and / or in situ hybridization (e.g., ISH). Two common IHC methods are available: direct assays and indirect assays. According to the first assay, the binding of an antibody to a target antigen is directly determined. This direct assay uses a labeling reagent, such as a fluorescent tag or an enzyme-labeled primary antibody, which can be visualized without further antibody interaction. In a typical indirect assay, an unconjugated primary antibody binds to the antigen, and then a labeled secondary antibody binds to the primary antibody. If the secondary antibody is conjugated to an enzymatic label, a chromogenic or fluorogenic substrate is added to visualize the antigen. Signal amplification occurs because several secondary antibodies can react with different epitopes on the primary antibody.

[0299] The primary and / or secondary antibodies used in IHC are typically labeled with a detectable moiety. Numerous labels are available, which can generally be grouped into the following categories: (a) 35 S, 14 C. 125 I, 3 H, 131(b) radioisotopes such as I; (b) colloidal gold particles; (c) fluorescent labels and / or derivatives of any one or more of the above, including, but not limited to, rare earth chelates (europium chelates), Texas Red, rhodamine, fluorescein, dansyl, Lissamine, umbelliferone, phycoerythrin, phycocyanin, or commercially available fluorophores such as SPECTRUM ORANGE 7 and SPECTRUM GREEN 7; (d) a variety of enzyme-substrate labels are available; U.S. Pat. No. 4,275,149 provides a review of some of these. Examples of enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase, see, e.g., U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidases such as horseradish peroxidase (HRPO), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, sugar oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like.

[0300] Examples of enzyme-substrate combinations include, for example, horseradish peroxidase (HRPO), which uses hydrogen peroxide as a substrate, alkaline phosphatase (AP), which uses para-nitrophenyl phosphate as a chromogenic substrate, and β-D-galactosidase (β-D-Gal), which uses a chromogenic substrate (e.g., p-nitrophenyl-β-D-galactosidase) or a fluorogenic substrate (e.g., 4-methylumbelliferyl-β-D-galactosidase). For a general review, see, e.g., U.S. Patent Nos. 4,275,149 and 4,318,980.

[0301] Specimens may be prepared, for example, manually or using an automated staining instrument (e.g., a Ventana BenchMark XT or Benchmark ULTRA instrument). Specimens prepared in this manner may be mounted and coverslipped. Slide evaluation may then be determined, for example, using a microscope, and staining intensity criteria routinely used in the art may be used. In one embodiment, when IHC is used to examine cells and / or tissues from a tumor, it is understood that staining is generally determined or assessed in the tumor cell(s) and / or tissue (versus stroma or surrounding tissue that may be present in the sample). In some embodiments, when IHC is used to examine cells and / or tissues from a tumor, it is understood that staining includes determining or assessing in tumor-infiltrating immune cells, including intratumoral or peritumoral immune cells. In some embodiments, the presence of the biomarker is detected by IHC in more than 0% of the samples, at least 1% of the samples, at least 5% of the samples, at least 10% of the samples, at least 15% of the samples, at least 15% of the samples, at least 20% of the samples, at least 25% of the samples, at least 30% of the samples, at least 35% of the samples, at least 40% of the samples, at least 45% of the samples, at least 50% of the samples, at least 55% of the samples, at least 60% of the samples, at least 65% of the samples, at least 70% of the samples, at least 75% of the samples, at least 80% of the samples, at least 85% of the samples, at least 90% of the samples, at least 95% of the samples, or more. Samples can be scored using any method known in the art, for example, by a pathologist or by automated image analysis.

[0302] In some embodiments of any of the methods, the biomarker is detected by immunohistochemistry using a diagnostic antibody (i.e., a primary antibody). In some embodiments, the diagnostic antibody specifically binds to a human antigen. In some embodiments, the diagnostic antibody is a non-human antibody. In some embodiments, the diagnostic antibody is a rat, mouse, or rabbit antibody. In some embodiments, the diagnostic antibody is a rabbit antibody. In some embodiments, the diagnostic antibody is a monoclonal antibody. In some embodiments, the diagnostic antibody is directly labeled. In other embodiments, the diagnostic antibody is indirectly labeled.

[0303] In some embodiments of any of the foregoing embodiments, the sample is obtained from the individual prior to administration of the anti-cancer treatment (e.g., minutes, hours, days, weeks (e.g., 1, 2, 3, 4, 5, 6, or 7 weeks), months, or years). In some embodiments of any of the foregoing methods, the sample from the individual is obtained about 2 to about 10 weeks (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks) after administration of the anti-cancer treatment. In some embodiments, the sample from the individual is obtained about 4 to about 6 weeks after administration of the anti-cancer treatment.

[0304] In some embodiments of any of the aforementioned methods, the expression level or number of biomarkers is detected in tissue samples, primary or cultured cells or cell lines, cell supernatants, cell lysates, platelets, serum, plasma, vitreous humor, lymphatic fluid, synovial fluid, follicular fluid, semen, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, tumor lysates, and tissue culture media, tissue extracts, such as homogenized tissues, tumor tissues, cell extracts, or any combination thereof. In some embodiments, the sample is a tissue sample (e.g., a tumor tissue sample), a cell sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof. In some embodiments, the tumor tissue sample contains tumor cells, tumor-infiltrating immune cells, stromal cells, or a combination thereof. In some embodiments, the tumor tissue sample is a formalin-fixed and paraffin-embedded (FFPE) sample, an archival sample, a fresh sample, or a frozen sample.

[0305] For example, in some embodiments of any of the aforementioned methods, the expression level of the biomarker is detected in tumor-infiltrating immune cells, tumor cells, PBMCs, or a combination thereof using known techniques (e.g., flow cytometry or IHC). Tumor-infiltrating immune cells include, but are not limited to, intratumoral immune cells, peritumoral immune cells, or any combination thereof, and other tumor stromal cells (e.g., fibroblasts). Such tumor-infiltrating immune cells include T lymphocytes (e.g., CD8 + T lymphocytes (e.g., CD8 + T Effector (T eff ) cells) and / or CD4 + T lymphocytes (e.g., CD4 + T eff The biomarkers may be erythrocytes (e.g., leukocytes, eosinophils, basophils), B lymphocytes, or other myeloid lineage cells, such as granulocytes (neutrophils, eosinophils, basophils), monocytes, macrophages, dendritic cells (e.g., interdigitating dendritic cells), histiocytes, and natural killer (NK) cells. In some embodiments, staining for the biomarker is detected as membrane staining, cytoplasmic staining, or a combination thereof. In other embodiments, the absence of a biomarker is detected as absence or no staining in the sample compared to a reference sample.

[0306] In certain embodiments of any of the aforementioned methods, the expression level of the biomarker is assessed in a sample containing or suspected of containing cancer cells. The sample may be, for example, a tissue biopsy or metastatic lesion obtained from a patient suffering from, suspected of, or diagnosed with cancer (e.g., kidney cancer, particularly renal cell carcinoma (RCC), e.g., advanced RCC or metastatic RCC (mRCC)). In some embodiments, the sample is kidney tissue, a kidney tumor biopsy, a known or suspected metastatic kidney cancer lesion or section, or a blood sample, e.g., a peripheral blood sample, known or suspected to contain circulating cancer cells, e.g., kidney cancer cells. The sample may contain both cancer cells (i.e., tumor cells) and non-cancerous cells (e.g., lymphocytes such as T cells or NK cells), and in certain embodiments, contains both cancerous and non-cancerous cells. Methods for obtaining biological samples, including tissue sections, biopsies, and bodily fluids (e.g., blood samples containing cancer / tumor cells), are well known in the art.

[0307] In some embodiments of any of the foregoing methods, the patient has carcinoma, lymphoma, blastoma (including medulloblastoma and retinoblastoma), sarcoma (including liposarcoma and synovial cell sarcoma), neuroendocrine tumors (including carcinoid tumor, gastrinoma, and islet cell carcinoma), mesothelioma, schwannoma (including acoustic neuroma), meningioma, adenocarcinoma, melanoma, and leukemia or lymphoid malignancies. In some embodiments, the cancer is selected from the group consisting of kidney cancer (e.g., renal cell carcinoma (RCC), e.g., advanced RCC or metastatic RCC (mRCC)), squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (including small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung, and squamous cell carcinoma of the lung), cancer of the peritoneum, hepatocellular carcinoma, gastric cancer or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer (e.g., HCC), liver cancer, breast cancer (including metastatic breast cancer), bladder cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, Merkel cell carcinoma, mycosis fungoides, testicular cancer, esophageal ... Cancer of the esophagus, bile duct tumors, head and neck cancer, 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-dividing cell NHL, bulky disease NHL, mantle cell lymphoma, AIDS-related lymphoma, and Waldenström's macroglobulinemia), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myeloblastic leukemia, a...

Claims

1. 1. A method of treating an individual having renal cancer, comprising administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist, wherein the individual has been identified as likely to benefit from the anti-cancer therapy based on the individual having sarcomatoid renal cancer.

2. 1. A method of treating an individual having renal cancer, comprising: (a) determining whether the individual has sarcomatoid renal cancer, wherein the presence of sarcomatoid renal cancer indicates that the individual may benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist; and (b) based on the presence of sarcomatoid renal cancer, administering to the individual an anti-cancer treatment comprising an effective amount of a VEGF antagonist and a PD-L1 axis binding antagonist; A method comprising:

3. 1. A method of identifying an individual having renal cancer who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist, comprising determining whether the individual has sarcomatoid renal cancer, wherein the presence of sarcomatoid renal cancer identifies the individual as an individual who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist.

4. 1. A method for selecting a treatment for an individual with kidney cancer, comprising: (a) determining whether the individual has sarcomatoid renal cancer, wherein the presence of sarcomatoid renal cancer identifies the individual as an individual who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist; and (b) selecting an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist based on the presence of sarcomatoid renal cancer; A method comprising:

5. 5. The method of claim 3 or 4, further comprising administering to the individual an effective amount of an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist.

6. The method of any one of claims 1 to 5, wherein the presence of sarcomatoid renal cancer is assessed by histological analysis of a sample obtained from the individual.

7. 7. The method of claim 6, wherein the kidney cancer is sarcomatoid if a tumor sample from the individual contains a focus(s) of high-grade malignant spindle cells of any component relative to the overall tumor area.

8. 8. The method of claim 6 or 7, wherein the spindle cells show moderate to marked atypia and / or resemble any form of sarcoma.

9. 9. The method of claim 7 or 8, wherein the spindle cells show evidence of epithelial differentiation as assessed by immunohistological positivity for keratin or epithelial membrane antigen (EMA).

10. 9. The method of claim 7 or 8, wherein the kidney cancer is renal cell carcinoma and the tumor sample has epithelial differentiation with concurrent areas of renal cell carcinoma.

11. 11. The method of any one of claims 1 to 10, wherein said benefit relates to improved progression free survival (PFS), overall survival (OS), overall response rate (ORR), complete response (CR) rate, or disease progression free rate (DFR).

12. 12. The method of claim 11, wherein the benefit relates to improved PFS.

13. The method of claim 11 , wherein the benefit relates to an improved OS.

14. 12. The method of claim 11, wherein the benefit relates to improved ORR.

15. The method of claim 11 , wherein the benefit relates to an improved CR rate.

16. The method of claim 11 , wherein the benefit relates to improved DFR.

17. 17. The method of claim 16, wherein DFR is determined as the time from initiation of treatment to the individual's first 2 or more point increase above baseline on the M.D. Anderson Symptom Index (MDASI) Interference scale.

18. The method of any one of claims 1 to 17, wherein the individual has a low or intermediate Memorial Sloan Kettering Cancer Center (MSKCC) risk score.

19. 1. A method of treating an individual having renal cancer, comprising administering to the individual an effective amount of an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist, wherein the individual has been identified as likely to benefit from the anti-cancer therapy based on having a low or intermediate MSKCC risk score.

20. 1. A method of treating an individual having renal cancer, comprising: (a) determining an MSKCC risk score for the individual, wherein a low or moderate MSKCC risk score indicates that the individual is likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist based on the individual having a low or intermediate MSKCC risk score; A method comprising:

21. 1. A method of identifying an individual having renal cancer who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist, comprising determining the individual's MSKCC risk score, wherein a low or moderate MSKCC risk score identifies the individual as likely to benefit from an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist.

22. 1. A method for selecting a treatment for an individual with kidney cancer, comprising: (a) determining an MSKCC risk score for the individual, wherein a low or moderate MSKCC risk score identifies the individual as likely to benefit from an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist; and (b) selecting an anti-cancer treatment comprising a VEGF antagonist and a PD-L1 axis binding antagonist based on the individual having a low or intermediate MSKCC risk score; A method comprising:

23. The individual has the following characteristics: (i) time from nephrectomy to systemic treatment less than 1 year, lack of nephrectomy, or initial diagnosis of metastatic disease; (ii) hemoglobin levels below the lower limit of normal (LLN) (in some cases, the normal range for hemoglobin is 13.5-17.5 g / dL for men and 12-15.5 g / dL for women); (iii) a corrected serum calcium level of greater than 10 mg / dL (optionally, the corrected serum calcium level is the serum calcium level (mg / dL) + 0.8 (4 - serum albumin (g / dL))); (iv) serum lactate dehydrogenase (LDH) levels greater than 1.5 times the upper limit of normal (ULN); in some cases, the ULN is 140 U / L; and / or (v) Karnofsky Performance Status (KPS) score less than 80; The method of any one of claims 18 to 22, wherein the individual has a low MSKCC risk score if they have three or more of:

24. The individual has the following characteristics: (i) time from nephrectomy to systemic treatment less than 1 year, lack of nephrectomy, or initial diagnosis of metastatic disease; (ii) hemoglobin levels below LLN (in some cases, the normal range for hemoglobin is 13.5-17.5 g / dL for men and 12-15.5 g / dL for women); (iii) a corrected serum calcium level of greater than 10 mg / dL (optionally, the corrected serum calcium level is the serum calcium level (mg / dL) + 0.8 (4 - serum albumin (g / dL))); (iv) a serum LDH level greater than 1.5 times the ULN (optionally, the ULN is 140 U / L); and / or (v) KPS score less than 80; The method of any one of claims 18 to 22, wherein the individual has an intermediate MSKCC risk score if they have one or two of:

25. The method of any one of claims 19 to 24, wherein the individual has sarcomatoid renal cancer.

26. The method of any one of claims 19 to 25, wherein said benefit relates to improved PFS, OS, ORR, CR rate, or DFR.

27. 27. The method of claim 26, wherein the benefit relates to improved PFS.

28. 27. The method of claim 26, wherein the benefit relates to an improved OS.

29. 27. The method of claim 26, wherein the benefit relates to improved ORR.

30. 27. The method of claim 26, wherein the benefit relates to an improved CR rate.

31. 27. The method of claim 26, wherein the benefit relates to improved DFR.

32. 32. The method of claim 31, wherein DFR is determined as the time from initiation of treatment to the individual's first 2 or more point increase above baseline on the MDASI Disability scale.

33. In a sample from the individual, the following genes: CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2; VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34; or IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2, The method of any one of claims 1 to 32, further comprising determining the expression level of one or more of:

34. (i) an expression level of one or more of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample that is equal to or greater than a reference expression level of the one or more genes; or (ii) an expression level of one or more of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34; or IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample that is less than a reference expression level of the one or more genes; identifies the individual as one who may benefit from treatment with an anti-cancer therapy comprising a VEGF antagonist and a PD-L1 axis binding antagonist.

35. 35. The method of claim 33 or 34, wherein the expression level of one or more of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample is determined to be equal to or greater than a reference level of the one or more genes.

36. 36. The method of claim 35, wherein expression levels of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen, at least nineteen, or all twenty of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, PD-L1, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample are determined to be equal to or greater than a reference level of the one or more genes.

37. The method of claim 35 or 36, wherein the expression level of one or more of CD8A, EOMES, PRF1, IFNG, or PD-L1 in the sample is determined to be equal to or greater than a reference level of the one or more genes.

38. 38. The method of claim 37, wherein the expression levels of CD8A, EOMES, PRF1, IFNG, and PD-L1 in the sample are determined to be equal to or greater than a reference level of CD8A, EOMES, PRF1, IFNG, and PD-L1.

39. 39. The method of any one of claims 33 to 38, wherein the expression level of one or more of IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample is determined to be equal to or greater than a reference level of the one or more genes.

40. 40. The method of claim 39, wherein the expression levels of at least one, at least two, at least three, at least four, at least five, or all six of IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample are determined to be equal to or greater than a reference level of the one or more genes.

41. 41. The method of claim 39 or 40, wherein the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, and PTGS2 in the sample are determined to be equal to or greater than the reference levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, and PTGS2.

42. 42. The method of any one of claims 33-41, wherein the expression level of PD-L1 in the sample is determined to be equal to or greater than a reference expression level of PD-L1, and the expression level of one or more additional genes selected from the group consisting of CD8A, EOMES, GZMA, GZMB, PRF1, IFNG, CXCL9, CXCL10, CXCL11, CD27, FOXP3, PD-1, CTLA4, TIGIT, IDO1, PSMB8, PSMB9, TAP1, or TAP2 in the sample is determined to be equal to or greater than a reference expression level of the one or more additional genes.

43. The method of claim 33 or 34, wherein the expression level of one or more of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 in the sample is determined to be less than a reference level of the one or more genes.

44. The method of claim 43, wherein the expression levels of at least one, at least two, at least three, at least four, at least five, at least six, or all seven of VEGFA, KDR, ESM1, PECAM1, FLT1, ANGPTL4, or CD34 in the sample are determined to be less than a reference level of the one or more genes.

45. The method of claim 43 or 44, wherein the expression level of one or more of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, or CD34 in the sample is determined to be less than a reference level of the one or more genes.

46. 46. ​​The method of claim 45, wherein the expression levels of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34 in the sample are determined to be less than the reference levels of VEGFA, KDR, ESM1, PECAM1, ANGPTL4, and CD34.

47. 35. The method of claim 33 or 34, wherein the expression level of one or more of IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample is determined to be less than a reference level of the one or more genes.

48. 48. The method of claim 47, wherein the expression levels of at least one, at least two, at least three, at least four, at least five, or all six of IL6, CXCL1, CXCL2, CXCL3, CXCL8, or PTGS2 in the sample are determined to be less than a reference level of the one or more genes.

49. 49. The method of claim 47 or 48, wherein the expression levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, and PTGS2 in the sample are determined to be less than the reference levels of IL6, CXCL1, CXCL2, CXCL3, CXCL8, and PTGS2.

50. 50. The method of any one of claims 34-49, wherein said reference levels of said one or more genes are determined from a population of individuals with renal cancer.

51. 51. The method of claim 50, wherein the reference level of the one or more genes is a median expression level determined in a population of patients with renal cancer.

52. 52. The method of claim 51 , wherein the reference level is the median Z-score of the normalized expression levels of the one or more genes.

53. The method of any one of claims 33 to 52, wherein the expression level is a nucleic acid expression level.

54. 54. The method of claim 53, wherein the nucleic acid expression level is an mRNA expression level.

55. 55. The method of claim 54, wherein the mRNA expression levels are determined by RNA-seq, RT-qPCR, qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, MassARRAY technology, ISH, or a combination thereof.

56. The method of any one of claims 33 to 52, wherein the expression level is a protein expression level.

57. 57. The method of claim 56, wherein the protein expression level is determined by immunohistochemistry (IHC), Western blot, enzyme-linked immunosorbent assay (ELISA), immunoprecipitation, immunofluorescence, radioimmunoassay, or mass spectrometry.

58. 58. The method of any one of claims 6 or 33-57, wherein the sample is a tissue sample, a cell sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

59. 59. The method of claim 58, wherein the tissue sample is a tumor tissue sample.

60. 60. The method of claim 59, wherein the tumor tissue sample comprises tumor cells, tumor-infiltrating immune cells, stromal cells, or a combination thereof.

61. 61. The method of claim 59 or 60, wherein the tumor tissue sample is a formalin-fixed and paraffin-embedded (FFPE) sample, an archival sample, a fresh sample, or a frozen sample.

62. The method of any one of claims 1-61, wherein the individual has not been previously treated for renal cancer.

63. 63. The method of any one of claims 1 to 62, wherein the kidney cancer is renal cell carcinoma (RCC).

64. 64. The method of claim 63, wherein the RCC is clear cell RCC.

65. The method of claim 63 or 64, wherein the RCC is locally advanced or metastatic RCC (mRCC).

66. 66. The method of any one of claims 1-65, wherein a tumor sample obtained from the patient has been determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that constitute about 1% or more of the tumor sample.

67. 67. The method of claim 66, wherein the tumor sample has been determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise between about 1% and less than 5% of the tumor sample.

68. 67. The method of claim 66, wherein the tumor sample has been determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise about 5% or more of the tumor sample.

69. 69. The method of claim 68, wherein the tumor sample has been determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise greater than about 5% and less than about 10% of the tumor sample.

70. 69. The method of claim 66 or 68, wherein the tumor sample obtained from the patient has been determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that constitute about 10% or more of the tumor sample.

71. 66. The method of any one of claims 1-65, wherein a tumor sample obtained from the patient has been determined to have a detectable expression level of PD-L1 in tumor-infiltrating immune cells that comprise less than about 1% of the tumor sample.

72. The method of any one of claims 1 to 71, wherein the VEGF antagonist is an anti-VEGF antibody or a VEGF receptor (VEGFR) inhibitor.

73. 73. The method of claim 72, wherein the VEGF antagonist is an anti-VEGF antibody.

74. 74. The method of claim 73, wherein the anti-VEGF antibody is bevacizumab.

75. 73. The method of claim 72, wherein the VEGF antagonist is a VEGFR inhibitor.

76. 76. The method of claim 75, wherein the VEGFR inhibitor is a multi-targeted tyrosine kinase inhibitor.

77. 77. The method of claim 76, wherein the multi-targeted tyrosine kinase inhibitor is sunitinib, axitinib, pazopanib, or cabozantinib.

78. 78. The method of claim 77, wherein the multi-targeted tyrosine kinase inhibitor is sunitinib.

79. 79. The method of any one of claims 1-78, wherein the PD-L1 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. 80. The method of claim 79, wherein the PD-L1 axis binding antagonist is a PD-L1 binding antagonist.

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

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

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

84. The method of any one of claims 81 to 83, wherein the PD-L1 binding antagonist inhibits binding of PD-L1 to both PD-1 and B7-1.

85. The method of any one of claims 80 to 84, wherein the PD-L1 binding antagonist is an anti-PD-L1 antibody.

86. 86. The method of claim 85, wherein the anti-PD-L1 antibody is selected from the group consisting of MPDL3280A (atezolizumab), YW243.55.S70, MDX-1105, MEDI4736 (durvalumab), and MSB0010718C (avelumab).

87. The anti-PD-L1 antibody has the following hypervariable regions (HVRs): (a) the HVR-H1 sequence of GFTFSDSWIH (SEQ ID NO:63); (b) the HVR-H2 sequence of AWISPYGGSTYYADSVKG (SEQ ID NO:64); (c) the HVR-H3 sequence of RHWPGGFDY (SEQ ID NO:65); (d) the HVR-L1 sequence of RASQDVSTAVA (SEQ ID NO:66); (e) the HVR-L2 sequence of SASFLYS (SEQ ID NO:67); and (f) the HVR-L3 sequence of QQYLYHPAT (SEQ ID NO:68); 87. The method of claim 85 or 86, comprising:

88. The anti-PD-L1 antibody, (a) a heavy chain variable (VH) domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 69); (b) a light chain variable (VL) domain comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 70); or (c) a VH domain as in (a) and a VL domain as in (b); The method of any one of claims 85 to 87, comprising:

89. The anti-PD-L1 antibody, (a) a VH domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:69; (b) a VL domain comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain as in (a) and a VL domain as in (b); 89. The method of claim 88, comprising:

90. The anti-PD-L1 antibody, (a) a VH domain comprising an amino acid sequence having at least 96% sequence identity to the amino acid sequence of SEQ ID NO: 69; (b) a VL domain comprising an amino acid sequence having at least 96% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain as in (a) and a VL domain as in (b); 90. The method of claim 89, comprising:

91. The anti-PD-L1 antibody, (a) a VH domain comprising an amino acid sequence having at least 97% sequence identity to the amino acid sequence of SEQ ID NO:69; (b) a VL domain comprising an amino acid sequence having at least 97% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain as in (a) and a VL domain as in (b); 91. The method of claim 90, comprising:

92. The anti-PD-L1 antibody, (a) a VH domain comprising an amino acid sequence having at least 98% sequence identity to the amino acid sequence of SEQ ID NO:69; (b) a VL domain comprising an amino acid sequence having at least 98% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain as in (a) and a VL domain as in (b); 92. The method of claim 91, comprising:

93. The anti-PD-L1 antibody, (a) a VH domain comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO:69; (b) a VL domain comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain as in (a) and a VL domain as in (b); 93. The method of claim 92, comprising:

94. The anti-PD-L1 antibody, (a) a VH domain comprising the amino acid sequence of SEQ ID NO:69; (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 70; or (c) a VH domain as in (a) and a VL domain as in (b); 94. The method of claim 93, comprising:

95. The anti-PD-L1 antibody, (a) a VH domain comprising the amino acid sequence of SEQ ID NO: 69; and (b) a VL domain comprising the amino acid sequence of SEQ ID NO: 70; 95. The method of claim 94, comprising:

96. 96. The method of claim 95, wherein the anti-PD-L1 antibody is atezolizumab.

97. 97. The method of any one of claims 1-96, wherein the PD-L1 axis binding antagonist is atezolizumab and the VEGF antagonist is bevacizumab.

98. 98. The method of claim 97, wherein the atezolizumab is administered intravenously at a dose of about 1200 mg every three weeks.

99. 99. The method of claim 97 or 98, wherein the bevacizumab is administered intravenously at a dose of about 15 mg / kg every three weeks.

100. 100. The method of any one of claims 1 to 99, further comprising administering to the individual an additional therapeutic agent.

101. 101. The method of claim 100, wherein the additional therapeutic agent is selected from the group consisting of an immunotherapeutic agent, a cytotoxic agent, a growth inhibitory agent, a radiotherapeutic agent, an anti-angiogenic agent, and combinations thereof.

102. The method of any one of claims 1 to 101, wherein the individual is a human.

103. 1. A pharmaceutical composition comprising a PD-L1 axis binding antagonist for use in treating an individual having renal cancer, said treatment comprising administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist, said individual being identified as likely to benefit from said anti-cancer treatment by virtue of having sarcomatoid renal cancer.

104. 1. A pharmaceutical composition comprising a PD-L1 axis binding antagonist for use in treating an individual having renal cancer, said treatment comprising administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist, said individual being identified as likely to benefit from said anti-cancer treatment based on having a low or intermediate MSKCC risk score.

105. 1. Use of a PD-L1 axis binding antagonist in the manufacture of a medicament for the treatment of an individual having renal cancer, said treatment comprising administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist, wherein said individual is identified as likely to benefit from said anti-cancer treatment based on having sarcomatoid renal cancer.

106. 1. Use of a PD-L1 axis binding antagonist in the manufacture of a medicament for the treatment of an individual having renal cancer, said treatment comprising administration of a PD-L1 axis binding antagonist in combination with a VEGF antagonist, wherein said individual is identified as likely to benefit from said anti-cancer treatment based on having a low or intermediate MSKCC risk score.

107. The pharmaceutical composition for use according to claim 103 or 104, or the use according to claim 105 or 106, wherein the benefit relates to improved progression-free survival (PFS), overall survival (OS), overall response rate (ORR), complete response (CR) rate, or disease-free rate (DFR).

108. 108. The pharmaceutical composition for use or use according to claim 107, wherein said benefit relates to improved PFS.

109. 108. The pharmaceutical composition for use or use according to claim 107, wherein said benefit relates to improved OS.

110. 108. The pharmaceutical composition for use or use according to claim 107, wherein said benefit relates to improved ORR.

111. 108. The pharmaceutical composition for use or use according to claim 107, wherein said benefit relates to improved CR rate.

112. 108. The pharmaceutical composition for use or use according to claim 107, wherein said benefit relates to improved DFR.

113. 113. The pharmaceutical composition for use or use according to claim 112, wherein DFR is determined as the time from the start of treatment to the individual's first increase of 2 or more points above baseline on the M.D. Anderson Symptom Assessment Inventory (MDASI) Interference scale.

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