EP4680639A1 - Combination therapies with bi-specific anti-egfr/c-met antibodies and anti-pd-1 antibodies - Google Patents

Abstract

The present invention relates to combination therapies for modulating the tumor microenvironment and enhancing immune cell infiltration into the tumor microenvironment with bispecific anti-EGFR/c-Met antibodies in combination with PD- (L)1 axis inhibitors. The invention also relates to combination therapies for inhibition of both EGFR and MET signaling pathways in a tumor cell, and targeting of EGFR and MET expressing tumor cells for destruction by immune effector cells, such as natural killer cells and macrophages, through antibody-dependent cellular cytotoxicity (ADCC) and trogocytosis mechanisms, respectively.

Description

COMBINATION THERAPIES WITH BISPECIFIC ANTI-EGFR/C-MET ANTIBODIES AND ANTI-PD-1 ANTIBODIES

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U . S . Provisional Patent Application No . 63/451,781, filed March 13, 2023 and U.S. Provisional Patent Application No. 63/459,857, filed April 17, 2023, the disclosure of which is herein incorporated by reference in their entirety.

REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY [0002] The sequence listing of the present application is submitted electronically via The United States Patent and Trademark Center Patent Center as an XML formatted sequence listing with a file name “JBI6783WOPCTl_SL.xml”, creation date of March 12, 2024, and a size of 39 kilobytes (KB). This sequence listing submitted is part of the specification and is herein incorporated by reference in its entirety.

FIELD OF THE INVENTION

[0003] The present invention relates to combination therapies and methods for remodeling the tumor microenvironment, inhibiting EGFR and MET signaling, and treating solid tumors comprising bispecific anti-EGFR/c-Met antibodies and inhibition of the PD-(L)1 axis.

BACKGROUND OF THE INVENTION

[0004] Immune checkpoint inhibitors, such as PD-(L)1 axis inhibitors, are effective first- line therapy for solid tumors. However, low response rate and acquired resistance over time has led to the need for additional therapeutic options. Unmet needs exist for immunotherapy that can function to remodel the tumor microenvironment, thus increasing thereapeutic responses, for various types of cancers, including head and neck cancers and lung cancers, which currently show suboptimal response to therapies.

[0005] This invention addresses this unmet need.

SUMMARY OF THE INVENTION

[0006] In one embodiment, the invention relates to a method of treating a solid tumor or ameliorating cancer progression in a subject in need thereof, the method comprising administering to the subject: (a) a PD-(L)1 axis inhibitor; and (b) a bispecific anti- EGFR/c-Met antibody.

[0007] In one embodiment, the invention relates to a method of enhancing immune cell infiltration into a solid tumor in a subject in need thereof, the method comprising administering to the subject: (a) a PD-(L)1 axis inhibitor; and (b) a bispecific anti- EGFR/c-Met antibody.

[0008] In one embodiment, the invention relates to a method of reducing glycolysis or lactic acid production in the tumor microenvironment in a subject in need thereof, the method comprising administering to the subject: (a) a PD-(L)1 axis inhibitor; and (b) a bispecific anti-EGFR/c-Met antibody.

[0009] In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6 and a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12. In one embodiment, the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. In one embodiment, the bispecific anti-EGFR/c-Met antibody is an IgGl isotype. In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

[00010] In one embodiment, the PD-(L)1 axis inhibitor is an inhibitory antibody. In one embodiment, the PD-(L)1 axis inhibitor is an anti-PD-1 antibody. In one embodiment, the inhibitory antibody is cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, or SHR-1210. In one embodiment, the pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26. In one embodiment, the pembrolizumab antibody comprises a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO: 30. In one embodiment, the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36. In one embodiment, the cetrelimab antibody comprises a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

[00011] In one embodiment, the cancer is an EGFR, c-Met or EGFR and c-Met expressing cancer associated with a wild-type EGFR, an EGFR activating mutation, an EGFR gene amplification, increased levels of circulating HGF, a wild-type c-Met, a c-Met activating mutation, a c-Met gene amplification or a mutant KRAS, or any combination thereof. In one embodiment, the EGFR activating mutation comprises L718Q, G719A, G719X (X being any amino acid), L861X (X being any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitution, deletion of E746-A750, deletion of R748-P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Vai and Ala (SVA) between S768 and V769, insertion of Asn and Ser (NS) between P772 and H773, insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof. In one embodiment, the mutant KRAS comprises a G12V, G12C or G12A substitution.

[00012] In one embodiment, the method inceases immune cell infiltration into a solid tumor. In one embodiment, the immune cells are T cells, B cells, or natural killer cells. In one embodiment, the T cells comprise CD8+ T cells and CD4+ T cells. In one embodiment, the T cells comprise CD8+ T cells.

[00013] In one embodiment, the method inceases the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

[00014] In one embodiment, the subject is suspected to have or has an EGFR, c-Met or EGFR and c-Met expressing cancer. In one embodiment, the subject is resistant or has acquired resistance to treatment with a prior anti-cancer therapy. [00015] In one embodiment, the prior anti-cancer therapy is a kinase inhibitor. In one embodiment, the kinase inhibitor is an inhibitor of EGFR, c-Met, HER2, HER3, HER4, VEGFR or AXL. In one embodiment, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib. [00016] In one embodiment, the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC). In one embodiment, the cancer is resistant to treatment with PD-(L)1 axis inhibitors. In one embodiment, the cancer is head and neck squamous cell carcinoma (HNSCC). In one embodiment, the cancer is lung squamous cell carcinoma (LUSC). In one embodiment, the cancer is non-small cell lung cancer (NSCLC).

[00017] In one embodiment, the method further comprises administering one or more anticancer therapies to the subject.

[00018] In one embodiment, the one or more anti-cancer therapies comprise a kinase inhibitor. In one embodiment, the kinase inhibitor is an inhibitor of EGFR, c- Met, HER2, HER3, HER4, VEGFR or AXL. In one embodiment, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

[00019] In one embodiment, the prior anti-cancer therapy is chemotherapy. In one embodiment, the prior anti-cancer therapy is a targeted anti-cancer therapy.

[00020] In one embodiment, the one or more anti-cancer therapies comprise chemotherapy. In one embodiment, the one or more anti-cancer therapies comprise a targeted anti-cancer therapy.

[00021] In one embodiment, the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered essentially concurrently to at two different injection sites. In one embodiment, the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are combined prior to administration and administered simultaneously. In one embodiment, the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered sequentially over the course of one or more days as part of a multi-step treatment regimen.

[00022] In one embodiment, the invention relates to a method of treating a solid tumor, ameliorating cancer progression, enhancing immune cell infiltration into a solid tumor, or reducing lactate production in the tumor microenvironment in a subject in need thereof, the method comprising administering a bispecific anti-EGFR/c-Met antibody to the subject, wherein the subject has received a prior administration of the PD-(L)1 axis inhibitor.

[00023] In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12. In one embodiment, the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. In one embodiment, the bispecific anti-EGFR/c-Met antibody is an IgGl isotype. In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

[00024] In one embodiment, the prior-administered PD-(L)1 axis inhibitor is an inhibitory antibody. In one embodiment, the prior-administered inhibitory antibody is pembrolizumab (KEYTRUDA®), cetrelimab, nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, or SHR-1210.

[00025] In one embodiment, the method enhances immune cell infiltration in the tumor or tumor microenvironment. In one embodiment, the immune cells are T cells, B cells, or natural killer cells. In one embodiment, the immune cells comprise CD8+ T cells and CD4+ T cells. In one embodiment, the immune cells comprise CD8+ T cells. In one embodiment, the method enhances the population of central memory cytotoxic T cells within the microenvironment of the tumor.

[00026] In one embodiment, the invention relates to a method of reducing lactate production in the tumor microenvironment in a subject in need thereof, the method comprising administering a combination of inhibitory agents to the subject, wherein the combination of inhibitory agents is (a) a combination comprising an inhibitor of EGFR, an inhibitor of c-Met and a PD-(L)1 axis inhibitor; or (b) a combination comprising an inhibitor of EGFR and an inhibitor of c-Met, wherein the subject has received a prior administration of a PD-(L)1 axis inhibitor.

[00027] In one embodiment, the combination of an inhibitor of EGFR and an inhibitor of c-Met comprises a bispecific anti-EGFR/c-Met antibody. In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12. In one embodiment, the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. In one embodiment, the bispecific anti-EGFR/c-Met antibody is an IgGl isotype. In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

[00028] In one embodiment, the PD-(L)1 axis inhibitor is an inhibitory antibody. In one embodiment, the PD-(L)1 axis inhibitor is an anti-PD-1 antibody. In one embodiment, the PD-(L)1 axis inhibitory antibody is cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, or SHR-1210. [00029] In one embodiment, the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

[00030] In one embodiment, the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC). In one embodiment, the cancer is resistant to treatment with PD-(L)1 axis inhibitors. In one embodiment, the cancer is head and neck squamous cell carcinoma (HNSCC). In one embodiment, the cancer is lung squamous cell carcinoma (LUSC). In one embodiment, the cancer is non-small cell lung cancer (NSCLC).

[00031] In one embodiment, the invention relates to a kit comprising a first pharmaceutical composition comprising a bispecific anti-EGFR/c-Met antibody and a second pharmaceutical composition comprising a PD-(L)1 axis inhibitor in two or more containers. In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises (a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and (b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12. In one embodiment, the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ IDNO: 16. In one embodiment, the bispecific anti-EGFR/c-Met antibody is an IgGl isotype. In one embodiment, the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

[00032] In one embodiment, the PD-(L)1 axis inhibitor is an inhibitory antibody. In one embodiment, the PD-(L)1 axis inhibitor is an anti-PD-1 antibody. In one embodiment, the inhibitory antibody is cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, or SHR-1210. In one embodiment, the pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26. In one embodiment, the pembrolizumab antibody comprises a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO: 30. In one embodiment, the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36. In one embodiment, the cetrelimab antibody comprises a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

[00033] In one embodiment, the kit comprises a first pharmaceutical composition comprising the bispecific anti-EGFR/c-Met antibody further comprises a first pharmaceutically acceptable excipient and a second pharmaceutical composition comprising the PD-(L)1 axis inhibitor further comprises a second pharmaceutically acceptable excipient.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts results from immunohistochemistry with anti-EGFR and anti-MET antibodies on head and neck squamous cell carcinoma (HNSCC) and lung squamous cell carcinoma (LUSC) humanized patient-derived xenograft (PDX) tumors.

FIG. 2A depicts a clinical profile of selected PDX models and experimental design. FIG. 2B shows that the LUSC model has primary resistance against pembrolizumab, showing consistent tumor growth despite treatment with pembrolizumab.

FIG. 3A depicts the anti-cancer effects of amivantamab and pembrolizumab combination treatment in HNSCC PDX model. FIG 3B shows the tumor progression of YHIM-3003 model over 19 days showing significant tumor regression by the combination treatment of amivantamab (30 mpk) and pembrolizumab (10 mpk) compared to single treatment of amivantamab and pembrolizumab (p<0.001). FIG. 4 depicts the anti-cancer effects of amivantamab and pembrolizumab combination treatment in LUSC PDX model.

FIG. 5 depicts data demonstrating persistent tumor regression after administration of amivantamab and in combination with pembrolizumab in LUSC PDX model.

FIG. 6 depicts images showing increased infiltrating cytotoxic T cells into the tumor nest after combination treatment.

FIG. 7 depicts a quantitative analysis of infiltrating cytotoxic T cells into the tumor nest after combination treatment; TME: tumor microenvironment, TN: tumor nest, ST: stroma.

FIG. 8A-8C depict data demonstrating that combination treatment enhanced central memory subset of cytotoxic T cells in tumor microenvironment of HNSCC. FIG. 8A shows heatmap of memory T cell subsets (central memory, effector memory and effector T cells) and activation markers in the tumor samples of HNSCC PDX. FIG. 8B shows heatmap of memory T cell subsets (central memory, effector memory and effector T cells) and activation markers in the tumor samples of YHIM-2010. FIG. 8C shows factors that combination of amivantamab and pembrolizumab positively affected in each humanized PDX model and both models shared enhancement of CD8+ T central memory subset by combination therapy.

FIG. 9 depicts data demonstrating that tumor reactive (CEA-stained) CD 8 T cells in HNSCC PDX tumor were abundant in the combination treatment group and were significantly higher in proportion compared to the control group (8.28 + 2.67 and 3.02 + 0.75, respectively, p<0.05). .

FIG. 10 depicts a single-cell (scRNA) analysis of immune populations using Azimuth clustering.

FIG. 11 depicts scRNA analysis of immune related transcripts using Azimuth clustering.

FIG. 12 depicts data demonstrating that treatment of pembrolizumab induced high level of EGFR and METexpressing tumor sub-cluster.

FIG. 13 depicts data demonstrating the EGFR/MET^hlgh and EGFR/MET^low tumor sub-clusters. Left panel: Tumor subcluster with elevated dual expression of EGFR and MET (EM^HIGH) was define and analysed for differentially expressed genes (DEGs).Right panel: EGFR and MET in EM^HIGH and EM^LOW, showing increased expression of both markers in EM^HIGH tumor cluster. FIG. 14 depicts data demonstrating that differentially expressed genes were analyzed in EGFR¹"⁸'¹' and EGFR^low expressing populations, showing genes related to immunomodulation, tumor metastasis, drug resistance and cancer sternness.

FIG. 15 depicts data demonstrating an analysis of hypoxia regulators and downstream mediators in EGFR¹”⁸¹¹ and EGFR^low expressing populations.

FIG. 16 depicts data demonstrating that the expression of EGFR was inversely correlated with infiltration of CD8+ T cells and NK cells in HNSCC and LUSC TCGA.

FIGs. 17A-17F depict data demonstrating that the expression of EGFR was inversely correlated with infiltration of CD8+ T cells and NK cells in HNSCC and LUSC TCGA. FIG. 17A shows correlation between expression of EGFR and biomarkers based on The Cancer Genome Atlas (TCGA) database. LDHA and SLC16A3 expression was positively correlated with expression of EGFR in both HNSCC and LUSC. FIG. 17B shows that expression of LDHA and SLC16A3 was significantly increased in EGFR^HIGH/MET^HIGH tumor subcluster (EM^HIGH) of HNSCC PDX (top). Additionally, expression of LDHA and SLC16A3 increased in pembrolizumab treated group. FIG. 17C shows that regulators of glycolysis (HK2, GPI, ALDO A, PGK1, PGAM1, ENO1, ENO2) comparatively increased in the EM^HIGH tumor subcluster (top) and pembrolizumab treated group (bottom). FIG. 17D shows that regulators of hypoxia (HIF1A, HDAC1, KDM1A, KDM2A) and downstream signalling markers (CA9, VEGFA, TWIST1) increased in the EM^HIGH tumor subcluster (top) and pembrolizumab treated group (bottom). FIG. 17E shows that H1703, LUSC human cancer cell, was treated with IFN-y for 24 hours to mimic the physiological response of pembrolizumab in the TME (left). Total protein and surface expression of EGFR and MET demonstrated strong correlation (right). FIG. 17F shows that protein expression of EGFR/p-EGFR, MET/p-MET, MCT4 (SLC16A3) and LDHA after 72 hours of amivantamab at 10 mg/ml in H1703. IFN-y was treated at 100 ng/ml for 24 hours.

FIG. 18 depicts data demonstrating changes in level of EGFR and MET expression in the tumor in different treatment groups.

FIG. 19 depicts data demonstrating changes in level of EGFR and MET expression in the tumor in different treatment groups.

FIGs. 20A-20C show that the upregulation of EGFR and MET in HNSCC PDX (YHIM-3003) tumor induced increased expression of immune checkpoints regulators in the EGFR^HIGH/MET^HIGH subcluster (EM^HIGH). FIG. 20A shows volcano plot of top 50 genes in EGFR^HIGH/MET^HIGH against EGFR^LOW/MET^LOW tumor subcluster analysed by log2 fold change (FC) against p-values. Red dots indicating transcripts with significantly increased fold changes including MET, PD-L1 and MET-regulated genes. FIG. 20B shows that expression of STAT-4/PD-L1 (MET, STAT4, CD274), MET regulated (BACE2, STK40, PRSS23, DPYD, CAV1, S100A4, PYGL) and MET-related immune checkpoints (HAVCR2, CD276) generally increased in the EGFR^HIGH/MET^HIGH tumor subcluster compared to the EGFR^LOW/MET^LOW subcluster. FIG. 20C shows that expression of MET-related markers in different treatment groups.

FIG. 21A depicts exemplary data demonstrating the effect on the body weight of YHIM-3003 PDX Mice. Treatments were initiated on day 0. Group body weights were presented as MEAN ± SEM (n = 10 mice per group). Data were graphed with all 10 mice remained on study. Body weights were recorded every other day. SEM, standard error of the mean.

FIG. 21B depicts exemplary data demonstrating the effect on the growth of YHIM-3003 tumors. Treatments were initiated on day 0. Tumor volumes were presented as group MEAN ± SEM (n = 10 mice per group). Groups were graphed with all 10 mice remained on study. Tumor volumes were measured every other day. SEM, standard error of the mean.

FIG. 22 A-B depicts the summary for combination synergy of amivantamab and pembrolizumab in EGFR^hl8hMET^hlgh cancer models.

FIGs. 23 A-B depict correlation of elevated EGFR/MET expression and poor immune response in anti-PD-1 non-responders from Gene Expression Omibus data.

DETAILED DESCRIPTION OF THE INVENTION

[00034] The invention is based, at least in part, on the finding that targeting EGFR/c-Met expressing cells with a bispecific anti-EGFR/c-Met antibody in combination with inhibition of the PD-(L)1 axis had the effect of remodeling the tumor immune microenvironment including reducing glycolysis and lactic acid production within the tumor microenvironment, increasing the levels of CD 8+ T cells in the tumor nest and increasing the population of central memory cytotoxic T cells within the tumor microenvironment. The invention is also based on the findings that the use of the bispecific anti-EGFR/c-Met antibodies of the invention (a) inhibited both EGFR and MET signaling pathways in a tumor cell, and (b) facilitated targeting of EGFR and MET expressing tumor cells for destruction by immune effector cells, such as natural killer cells and macrophages, through antibody -dependent cellular cytotoxicity (ADCC) and trogocytosis mechanisms, respectively. [00035] The identification of this mechanism provides basis for selecting patients for treatment who may benefit from combination therapies comprising an anti-EGFR/c-Met bispecific antibody and an inhibitor of the PD-(L)1 axis.

Definitions

[00036] All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as though fully set forth.

[00037] It is to be understood that the terminology used herein is for describing particular embodiments only and is not intended to be limiting. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

[00038] Although any methods and materials similar or equivalent to those described herein may be used in the practice for testing of the present invention, exemplary materials and methods are described herein. In describing and claiming the present invention, the following terminology will be used.

[00039] When a list is presented, unless stated otherwise, it is to be understood that each individual element of that list, and every combination of that list, is a separate embodiment. For example, a list of embodiments presented as “A, B, or C” is to be interpreted as including the embodiments, “A,” “B,” “C,” “A orB,” “A or C,” “B or C,” or “A, B, or C.”

[00040] As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to “a cell” includes a combination of two or more cells, and the like.

[00041] The conjunctive term “and/or” between multiple recited elements is understood as encompassing both individual and combined options. For instance, where two elements are conjoined by “and/or,” a first option refers to the applicability of the first element without the second. A second option refers to the applicability of the second element without the first. A third option refers to the applicability of the first and second elements together. Any one of these options is understood to fall within the meaning, and therefore satisfy the requirement of the term “and/or” as used herein. Concurrent applicability of more than one of the options is also understood to fall within the meaning, and therefore satisfy the requirement of the term “and/or.”

[00042] The transitional terms “comprising,” “consisting essentially of,” and “consisting of’ are intended to connote their generally accepted meanings in the patent vernacular; that is, (i) “comprising,” which is synonymous with “including,” “containing,” or “characterized by,” is inclusive or open-ended and does not exclude additional, unrecited elements or method steps; (ii) “consisting of’ excludes any element, step, or ingredient not specified in the claim; and (iii) “consisting essentially of’ limits the scope of a claim to the specified materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention. Embodiments described in terms of the phrase “comprising” (or its equivalents) also provide as embodiments those independently described in terms of “consisting of’ and “consisting essentially of.”

[00043] “Co-administration,” “administration with,” “administration in combination with,” “in combination with” or the like, encompass administration of the selected therapeutics or drugs to a single patient, and are intended to include treatment regimens in which the therapeutics or dmgs are administered by the same or different route of administration or at the same or different time.

[00044] “Isolated” refers to a homogenous population of molecules (such as synthetic polynucleotides, polypeptides vectors or viruses) which have been substantially separated and/or purified away from other components of the system the molecules are produced in, such as a recombinant cell, as well as a protein that has been subjected to at least one purification or isolation step. “Isolated” refers to a molecule that is substantially free of other cellular material and/or chemicals and encompasses molecules that are isolated to a higher purity, such as to 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% purity.

[00045] “Treat”, “treating” or “treatment” of a disease or disorder such as cancer refers to accomplishing one or more of the following: reducing the severity and/or duration of the disorder, inhibiting worsening of symptoms characteristic of the disorder being treated, limiting or preventing recurrence of the disorder in subjects that have previously had the disorder, or limiting or preventing recurrence of symptoms in subjects that were previously symptomatic for the disorder.

[00046] “Prevent”, “preventing”, “prevention”, or “prophylaxis” of a disease or disorder means preventing that a disorder occurs in subject.

[00047] “Diagnosing” or “diagnosis” refers to methods to determine if a subject is suffering from a given disease or condition or may develop a given disease or condition in the future or is likely to respond to treatment for a prior diagnosed disease or condition, i.e., stratifying a patient population on likelihood to respond to treatment. Diagnosis is typically performed by a physician based on the general guidelines for the disease to be diagnosed or other criteria that indicate a subject is likely to respond to a particular treatment.

[00048] “Responsive”, “responsiveness” or “likely to respond” refers to any kind of improvement or positive response, such as alleviation or amelioration of one or more symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable.

[00049] “Newly diagnosed” refers to a subject who has been diagnosed with EGFR or c- Met expressing cancer but has not yet received treatment for multiple myeloma.

[00050] “Therapeutically effective amount” refers to an amount effective, at doses and for periods of time necessary, to achieve a desired therapeutic result. A therapeutically effective amount may vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of a therapeutic or a combination of therapeutics to elicit a desired response in the individual. Exemplary indicators of an effective therapeutic or combination of therapeutics that include, for example, improved well-being of the patient.

[00051] “Subject” includes any human or nonhuman animal. “Nonhuman animal” includes all vertebrates, e.g., mammals and non-mammals, such as nonhuman primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. The terms “subject” and “patient” are used interchangeably herein.

[00052] “About” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. Unless explicitly stated otherwise within the Examples or elsewhere in the Specification in the context of a particular assay, result or embodiment, “about” means within one standard deviation per the practice in the art, or a range of up to 5%, whichever is larger.

[00053] “Cancer” refers to an abnormal growth of cells which tend to proliferate in an uncontrolled way and, in some cases, to metastasize (spread) to other areas of a patient’s body.

[00054] “EGFR or c-Met expressing cancer” refers to cancer that has detectable expression of EGFR or c-Met or has EGFR or c-Met mutation or amplification. EGFR or c-Met expression, amplification and mutation status can be detected using know methods, such as sequencing, fluorescent in situ hybridization, immunohistochemistry, flow cytometry or western blotting. [00055] “Epidermal growth factor receptor” or “EGFR” refers to the human EGFR (also known as HER1 orErbBl (Ullrich et al., Nature 309:418-425, 1984) having the amino acid sequence shown in GenBank accession number NP 005219, as well as naturally occurring variants thereof.

[00056] “Hepatocyte growth factor receptor” or “c-Mef ’ or “MET” as used herein refers to the human c-Met having the amino acid sequence shown in GenBank Accession No: NP 001120972 and natural variants thereof.

[00057] “Bispecific anti-EGFR/c-Met antibody” or “bispecific EGFR/c-Met antibody” refers to a bispecific antibody having a first domain that specifically binds EGFR and a second domain that specifically binds c-Met. The domains specifically binding EGFR and c-Met are typically VH/VL pairs, and the bispecific anti-EGFR/c-Met antibody is monovalent in terms of binding to EGFR and c-Met.

[00058] “Biosimilar” (of an approved reference product/biological drug, i.e., reference listed drug) refers to a biological product that is highly similar to the reference product notwithstanding minor differences in clinically inactive components with no clinically meaningful differences between the biosimilar and the reference product in terms of safety, purity and potency, based upon data derived from (a) analytical studies that demonstrate that the biological product is highly similar to the reference product notwithstanding minor differences in clinically inactive components; (b) animal studies (including the assessment of toxicity); and/or (c) a clinical study or studies (including the assessment of immunogenicity and pharmacokinetics or pharmacodynamics) that are sufficient to demonstrate safety, purity, and potency in one or more appropriate conditions of use for which the reference product is licensed and intended to be used and for which licensure is sought for the biosimilar. The biosimilar may be an interchangeable product that may be substituted for the reference product at the pharmacy without the intervention of the prescribing healthcare professional. To meet the additional standard of “interchangeability,” the biosimilar is to be expected to produce the same clinical result as the reference product in any given patient and, if the biosimilar is administered more than once to an individual, the risk in terms of safety or diminished efficacy of alternating or switching between the use of the biosimilar and the reference product is not greater than the risk of using the reference product without such alternation or switch. Further, the biosimilar must utilize the same mechanism(s) of action for the proposed conditions of use to the extent the mechanisms are known for the reference product. Also, the condition or conditions of use that were prescribed, recommended, or suggested in the labeling proposed for the biosimilar, have been previously approved for the reference product. The route of administration, the dosage form, and/or the strength of the biosimilar must also be the same as those of the reference product and the biosimilar must be manufactured, processed, packed or held in a facility that meets standards designed to assure that the biosimilar is safe, pure and efficacious. The biosimilar may include a one or more post- translational modifications what are different from the reference product, such as a different glycosylation profde, that is not expected to change the biosimilar performance. [00059] “Specific binding” or “specifically binds” or “specifically binding” or “binds” refer to an antibody binding to an antigen or an epitope within the antigen with greater affinity than for other antigens. Typically, the antibody binds to the antigen or the epitope within the antigen with an equilibrium dissociation constant (KD) of about 5x10-8 M or less, for example about 1x10-9 M or less, about 1x10-10 M or less, about 1x10-11 M or less, or about 1x10-12 M or less, typically with the KD that is at least one hundred-fold less than its KD for binding to a non-specific antigen (e.g., BSA, casein). The dissociation constant may be measured using known protocols. Antibodies that bind to the antigen or the epitope within the antigen may, however, have cross-reactivity to other related antigens, for example to the same antigen from other species (homologs), such as human or monkey, for example Macaca fascicularis (cynomolgus, cyno) or Pan troglodytes (chimpanzee, chimp). While a monospecific antibody binds one antigen or one epitope, a bispecific antibody binds two distinct antigens or two distinct epitopes.

[00060] “Antibodies” is meant in a broad sense and includes immunoglobulin molecules including monoclonal antibodies including murine, human, humanized and chimeric monoclonal antibodies, antigen binding fragments, multispecific antibodies, such as bispecific, trispecific, tetraspecific etc., dimeric, tetrameric or multimeric antibodies, single chain antibodies, domain antibodies and any other modified configuration of the immunoglobulin molecule that comprises an antigen binding site of the required specificity. “Full length antibodies” are comprised of two heavy chains (HC) and two light chains (LC) inter-connected by disulfide bonds as well as multimers thereof (e.g. IgM). Each heavy chain is comprised of a heavy chain variable region (VH) and a heavy chain constant region (comprised of domains CHI, hinge, CH2 and CH3). Each light chain is comprised of a light chain variable region (VL) and a light chain constant region (CL). The VH and the VL regions may be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with framework regions (FR). Each VH and VL is composed of three CDRs and four FR segments, arranged from amino-to-carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. [00061] “Complementarity determining regions” (CDR) are antibody regions that bind an antigen. CDRs may be defined using various delineations such as Kabat (Wu et al. (1970) J Exp Med 132: 211-50) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), Chothia (Chothia et al. (1987) J Mol Biol 196: 901-17), IMGT (Lefranc et al. (2003) Dev Comp Immunol 27: 55-77) and AbM (Martin and Thornton (1996) J Bmol Biol 263: SOO- 15). The correspondence between the various delineations and variable region numbering are described (see e.g. Lefranc et al. (2003) Dev Comp Immunol 27: 55-77; Honegger and Pluckthun, (2001) J Mol Biol 309:657-70; International ImMunoGeneTics (IMGT) database; Web resources, http://www_imgt_org). Available programs such as abYsis by UCL Business PLC may be used to delineate CDRs. The term “CDR”, “HCDR1”, “HCDR2”, “HCDR3”, “LCDR1”, “LCDR2” and “LCDR3” as used herein includes CDRs defined by any of the methods described supra, Kabat, Chothia, IMGT or AbM, unless otherwise explicitly stated in the specification

[00062] Immunoglobulins may be assigned to five major classes, IgA, IgD, IgE, IgG and IgM, depending on the heavy chain constant domain amino acid sequence. IgA and IgG are further sub-classified as the isotypes IgAl, IgA2, IgGl, IgG2, IgG3 and IgG4. Antibody light chains of any vertebrate species may be assigned to one of two clearly distinct types, namely kappa (K) and lambda (X), based on the amino acid sequences of their constant domains.

[00063] “Antigen binding fragment” refers to a portion of an immunoglobulin molecule that binds an antigen. Antigen binding fragments may be synthetic, enzymatically obtainable or genetically engineered polypeptides and include the VH, the VL, the VH and the VL, Fab, F(ab')2, Fd and Fv fragments, domain antibodies (dAb) consisting of one VH domain or one VL domain, shark variable IgNAR domains, camelized VH domains, minimal recognition units consisting of the amino acid residues that mimic the CDRs of an antibody, such as FR3-CDR3-FR4 portions, the HCDR1, the HCDR2 and/or the HCDR3 and the LCDR1, the LCDR2 and/or the LCDR3. VH and VL domains may be linked together via a synthetic linker to form various types of single chain antibody designs where the VH/VL domains may pair intramolecularly, or intermolecularly in those cases when the VH and VL domains are expressed by separate single chain antibody constmcts, to form a monovalent antigen binding site, such as single chain Fv (scFv) or diabody; described for example in Int. Patent Publ. Nos. W01998/44001, WO1988/01649, WO1994/13804 and W01992/01047. [00064] “Monoclonal antibody” refers to an antibody obtained from a substantially homogenous population of antibody molecules, i.e., the individual antibodies comprising the population are identical except for possible well-known alterations such as removal of C-terminal lysine from the antibody heavy chain or post-translational modifications such as amino acid isomerization or deamidation, methionine oxidation or asparagine or glutamine deamidation. Monoclonal antibodies typically bind one antigenic epitope. A bispecific monoclonal antibody binds two distinct antigenic epitopes. Monoclonal antibodies may have heterogeneous glycosylation within the antibody population. Monoclonal antibody may be monospecific or multispecific such as bispecific, monovalent, bivalent or multivalent.

[00065] “Humanized antibody” refers to an antibody in which at least one CDR is derived from non-human species and at least one framework is derived from human immunoglobulin sequences. Humanized antibody may include substitutions in the frameworks so that the frameworks may not be exact copies of expressed human immunoglobulin or human immunoglobulin germline gene sequences.

[00066] “Human antibody” refers to an antibody that is optimized to have minimal immune response when administered to a human subject. Variable regions of human antibody are derived from human immunoglobulin sequences. If human antibody contains a constant region or a portion of the constant region, the constant region is also derived from human immunoglobulin sequences. Human antibody comprises heavy and light chain variable regions that are “derived from” sequences of human origin if the variable regions of the human antibody are obtained from a system that uses human germline immunoglobulin or rearranged immunoglobulin genes. Such exemplary systems are human immunoglobulin gene libraries displayed on phage, and transgenic non-human animals such as mice or rats carrying human immunoglobulin loci. “Human antibody” typically contains amino acid differences when compared to the immunoglobulins expressed in humans due to differences between the systems used to obtain the human antibody and human immunoglobulin loci, introduction of somatic mutations or intentional introduction of substitutions into the frameworks or CDRs, or both. Typically, “human antibody” is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in amino acid sequence to an amino acid sequence encoded by human germline immunoglobulin or rearranged immunoglobulin genes. In some cases, “human antibody” may contain consensus framework sequences derived from human framework sequence analyses, for example as described in Knappik et al., (2000) J Mol Biol 296:57-86, or synthetic HCDR3 incorporated into human immunoglobulin gene libraries displayed on phage, for example as described in Shi et al., (2010) J Mol Biol 397:385-96, and in Int. Patent Publ. No. W02009/085462. Antibodies in which at least one CDR is derived from a non-human species are not included in the definition of “human antibody”

[00067] “Recombinant” refers to DNA, antibodies and other proteins that are prepared, expressed, created or isolated by recombinant means when segments from different sources are joined to produce recombinant DNA, antibodies or proteins.

[00068] “Bispecific” refers to an antibody that specifically binds two distinct antigens or two distinct epitopes within the same antigen. The bispecific antibody may have crossreactivity to other related antigens, for example to the same antigen from other species (homologs), such as human or monkey, for example Macaca cynomolgus (cynomolgus, cyno) or Pan troglodytes, or may bind an epitope that is shared between two or more distinct antigens.

[00069] “Multispecific” refers to an antibody that specifically binds two or more distinct antigens or two or more distinct epitopes within the same antigen. The multispecific antibody may have cross-reactivity to other related antigens, for example to the same antigen from other species (homologs), such as human or monkey, for example Macaca cynomolgus (cynomolgus, cyno) or Pan troglodytes, or may bind an epitope that is shared between two or more distinct antigens.

[00070] “Monocyte” refers to the CD14+CD34- mononuclear white cell, belonging to a type of white blood cell involved in first-line defensive mechanisms and is recognized as able to differentiate into a dendritic cell or macrophage precursor. Monocyes normally move in the blood system. In response to external stimulating signals, monocytes secrete many immunoregulator cytokines, move to the site of infenction in the tissue or to a site of tumor, and differentiate into macrophages. In particular, a monocyte expresses elevated levels of the CD 14 surface antigen marker, and may express at least one biomarker selected from CD64, CD93, CD180, CD328, CD329 or peanut agglutinin protein (PNA). [00071] “Enhance” or “induce” refers to potentiation of one or more function or activity of a macrophage by more than 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100%, or by a statistically significant manner when compared to a control (e.g., potentiation in the presence or absence of an agent that enhances macrophage activity).

[00072] "Agonist" refers to a molecule that, when bound to a cellular protein, induces at least one reaction or activity that is induced by a natural ligand of the protein. The molecule is an agonist when the at least one reaction or activity is induced by at least about 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% greater than the at least one reaction or activity induced in the absence of the agonist (e.g., negative control), or when the induction is statistically significant when compared to the induction in the absence of the agonist.

[00073] "Antagonist" or “inhibitor” refers to a molecule that, when bound to a cellular protein, suppresses at least one reaction or activity that is induced by a natural ligand of the protein. A molecule is an antagonist when the at least one reaction or activity is suppressed by at least about 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% more than the at least one reaction or activity suppressed in the absence of the antagonist (e.g., negative control), or when the suppression is statistically significant when compared to the suppression in the absence of the antagonist. [00074] “PD-(L)1 axis inhibitor” refers to a molecule that inhibits PD-1 downstream signaling. PD-(L)1 axis inhibitor may be a molecule that binds PD-1, PD-L1 or PD-L2. [00075] “PD-(L)1 axis inhibitor resistant” or “Anti-PD-(L)1 axis inhibitor resistant” refers to a cancer that did not respond to a treatment with a PD-(L)1 axis inhibitor.

[00076] “Biological sample” refers to a collection of similar fluids, cells, or tissues isolated from a subject, as well as fluids, cells, or tissues present within a subject. Exemplary samples are biological fluids such as blood, serum and serosal fluids, plasma, lymph, urine, saliva, cystic fluid, tear drops, feces, sputum, mucosal secretions of the secretory tissues and organs, vaginal secretions, ascites fluids, fluids of the pleural, pericardial, peritoneal, abdominal and other body cavities, fluids collected by bronchial lavage, synovial fluid, liquid solutions contacted with a subject or biological source, for example, cell and organ culture medium including cell or organ conditioned medium, lavage fluids and the like, tissue biopsies, tumor tissue biopsies, tumor tissue samples, fine needle aspirations, surgically resected tissue, organ cultures or cell cultures.

[00077] “Low fucose” or “low fucose content” as used in the application refers to antibodies with fucose content of about between 1 %- 15%.

[00078] “Normal fucose” or ‘normal fucose content” as used herein refers to antibodies with fucose content of about over 50%, typically about over 80% or over 85%.

Bispecific Anti-EGFR/c-Met Antibody

[00079] Bispecific anti-EGFR/c-Met antibodies used in the methods of the disclosure may be generated for example using Fab arm exchange (or half molecule exchange) between two monospecific bivalent antibodies by introducing substitutions at the heavy chain CH3 interface in each half molecule to favor heterodimer formation of two antibody half molecules having distinct specificity either in vitro in cell-free environment or using coexpression. The Fab arm exchange reaction is the result of a disulfide-bond isomerization reaction and dissociation-association of CH3 domains. The heavy chain disulfide bonds in the hinge regions of the parental monospecific antibodies are reduced. The resulting free cysteines of one of the parental monospecific antibodies form an inter heavy -chain disulfide bond with cysteine residues of a second parental monospecific antibody molecule and simultaneously CH3 domains of the parental antibodies release and reform by dissociation-association. The CH3 domains of the Fab arms may be engineered to favor heterodimerization over homodimerization. The resulting product is a bispecific antibody having two Fab arms or half molecules which each bind a distinct epitope, i.e. an epitope on EGFR and an epitope on c-Met. For example, the bispecific antibodies of the invention may be generated using the technology described in IntPat. Publ. No. WO2011/131746. Mutations F405L in one heavy chain and K409R in the other heavy chain may be used in case of IgGl antibodies. For IgG2 antibodies, a wild-type IgG2 and a IgG2 antibody with F405L and R409K substitutions may be used. For IgG4 antibodies, a wild-type IgG4 and a IgG4 antibody with F405L and R409K substitutions may be used. To generate bispecific antibodies, first monospecific bivalent antibody and the second monospecific bivalent antibody are engineered to have the aforementioned mutation in the Fc region, the antibodies are incubated together under reducing conditions sufficient to allow the cysteines in the hinge region to undergo disulfide bond isomerization; thereby generating the bispecific antibody by Fab arm exchange. The incubation conditions may optimally be restored to non-reducing. Exemplary reducing agents that may be used are 2- mercaptoethylamine (2-MEA), dithiothreitol (DTT), dithioerythritol (DTE), glutathione, tris(2-carboxyethyl)phosphine (TCEP), L-cysteine and beta- mercaptoethanol. For example, incubation for at least 90 min at a temperature of at least 20°C in the presence of at least 25 mM 2-MEA or in the presence of at least 0.5 mM dithiothreitol at a pH of from 5-8, for example at pH of 7.0 or at pH of 7.4 may be used.

[00080] In some embodiments, the bispecific anti-EGFR/c-Met antibody is an IgGl isotype. Some variations can exist within the IgGl constant domain (e.g. well-known allotypes), with variation at positions 214, 356, 358, 422, 431, 435 o 436 (residue numbering according to the EU numbering) (see e.g. IMGT Web resources; IMGT Repertoire (IG and TR); Proteins and alleles; allotypes). The bispecific anti-EGFR/c-Met antibody may be any IgGl allotype, such as Glml7, Glm3, Glml, Glm2, Glm27 or Glm28 or others, known in the art. [00081] Bispecific anti-EGFR/c-Met antibodies used in the methods of the disclosure may also be generated using designs such as the Knob-in-Hole (Genentech), CrossMAbs (Roche) and the electrostatically -matched (Chugai, Amgen, NovoNordisk, Oncomed), the LUZ-Y (Genentech), the Strand Exchange Engineered Domain body (SEEDbody)(EMD Serono), and the Biclonic (Merus).

[00082] In the “knob -in-hole” strategy (see, e.g., Inti. Publ. No. WO 2006/028936) select amino acids forming the interface of the CH3 domains in human IgG can be mutated at positions affecting CH3 domain interactions to promote heterodimer formation. An amino acid with a small side chain (hole) is introduced into a heavy chain of an antibody specifically binding a first antigen and an amino acid with a large side chain (knob) is introduced into a heavy chain of an antibody specifically binding a second antigen. After co-expression of the two antibodies, a heterodimer is formed as a result of the preferential interaction of the heavy chain with a “hole” with the heavy chain with a “knob”.

Exemplary CH3 substitution pairs forming a knob and a hole are (expressed as modified position in the first CH3 domain of the first heavy chain/ modified position in the second CH3 domain of the second heavy chain): T366Y/F405A, T366W/F405W, F405W/Y407A, T394W/Y407T, T394S/Y407A, T366W/T394S, F405W/T394S and T366W/T366S L368A Y407V.

[00083] CrossMAb technology, in addition to utilizing the “knob-in-hole” strategy to promoter Fab arm exchange utilizes CH1/CL domain swaps in one half arm to ensure correct light chain pairing of the resulting bispecific antibody (see e.g. U.S. Patent No. 8,242,247).

[00084] Other cross-over strategies may be used to generate full length bispecific antibodies of the invention by exchanging variable or constant, or both domains between the heavy chain and the light chain or within the heavy chain in the bispecific antibodies, either in one or both arms. These exchanges include for example VH-CH1 with VL-CL, VH with VL, CH3 with CL and CH3 with CHI as described in Int. Patent Publ. Nos. W02009/080254, W02009/080251, W02009/018386 and W02009/080252.

[00085] Other strategies such as promoting heavy chain heterodimerization using electrostatic interactions by substituting positively charged residues at one CH3 surface and negatively charged residues at a second CH3 surface may be used, as described in US Patent Publ. No. US2010/0015133; US Patent Publ. No. US2009/0182127; US Patent Publ. No. US2010/028637 or US Patent Publ. No. US2011/0123532. In other strategies, heterodimerization may be promoted by following substitutions (expressed as modified positions in the first CH3 domain of the first heavy chain/ modified position in the second CH3 domain of the second heavy chain): L351Y F405A Y407V/T394W,

T366I K392M T394 W/F405 A Y407 V, T366L K392M T394 W/F405 A Y407 V, L351Y Y407A/T366A K409F, L351Y_Y4O7A/T366V_K4O9F, Y407A/T366A K409F, or T350V_L351Y_F405A_Y407V/T350V_T366L_K392L_T394W as described in U.S.

Patent Publ. No. US2012/0149876 or U.S. Patent Publ. No. US2013/0195849.

[00086] SEEDbody technology may be utilized to generate bispecific antibodies of the invention. SEEDbodies have, in their constant domains, select IgG residues substituted with IgA residues to promote heterodimerization as described in U.S. Patent No. US20070287170.

[00087] Mutations are typically made at the DNA level to a molecule such as the constant domain of the antibody using standard methods.

[00088] In some embodiments, the bispecific anti-EGFR/c-Met antibody comprises [00089] a first domain that binds EGFR comprising a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and a second domain that binds c-Met comprising the HCDR1 of SEQ ID NO: 7, the HCDR2 of SEQ ID NO: 8, the HCDR3 of SEQ ID NO: 9, the LCDR1 of SEQ ID NO: 10, the LCDR2 of SEQ ID NO: 11 and the LCDR3 of SEQ ID NO: 12.

[00090] In some embodiments, the first domain that binds EGFR comprises a heavy chain variable domain (VH) of SEQ ID NO: 13 and a light chain variable domain (VL) of SEQ ID NO: 14; and the second domain that binds c-Met comprises the VH of SEQ ID NO: 15 and the VL of SEQ ID NO: 16.

[00091] In some embodiments, the bispecific anti-EGFR/c-Met antibody is an IgGl isotype. Some variation exists within the IgGl constant domain (e.g. well-known allotypes), with variation at positions 214, 356, 358, 422, 431, 435 o 436 (residue numbering according to the EU numbering) (see e.g. IMGT Web resources; IMGT Repertoire (IG and TR); Proteins and alleles; allotypes). The bispecific anti-EGFR/c-Met antibody may be of any IgGl allotype, such as Glml7, Glm3, Glml, Glm2, Glm27 or Glm28.

[00092] In some embodiments, the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20. [00093] Other bispecific anti-EGFR/c-Met antibodies publicly available may also be used in the methods of the disclosure as long as they demonstrate similar characteristics when compared to amivantamab as described in U.S. Pat. No. 9,593,164. Bispecific anti- EGFR/c-Met antibodies that may be used in the methods of the disclosure may also be generated by combining EGFR binding VH/VL domains and c-Met binding VH/VL domains that are publicly available and testing the resulting bispecific antibodies for their characteristics as described in U.S. Pat. No. 9,593,164. In some embodiments, the bispecific anti-EGFR/c-Met antibodies is a Biosimilar.

[00094] In some embodiments, the bispecific anti-EGFR/c-Met antibody has reduced fucose content of about between 1% to about 10%. The bispecific anti-EGFR/c-Met antibody having reduced fucose content may be more efficacious in the treatment of patients with FcyRIIIa-158F/F or FcyRIIIa-158F/V genotypes. Patients can be analyzed for their FcyRIIIa polymorphism using routine methods.

[00095] Antibodies with reduced fucose content can be made using different methods reported to lead to the successful expression of relatively high defucosylated antibodies bearing the biantennary complex-type of Fc oligosaccharides such as control of culture osmolality (Konno et al., Cytotechnology 64(:249-65, 2012), application of a variant CHO line Lecl3 as the host cell line (Shields et al., J Biol Chem 277:26733-26740, 2002), application of a variant CHO line EB66 as the host cell line (Olivier et al., MAbs ;2(4), 2010; Epub ahead of print; PMID:20562582), application of a rat hybridoma cell line YB2/0 as the host cell line (Shinkawa et al., J Biol Chem 278:3466-3473, 2003), introduction of small interfering RNA specifically against the a 1,6-fucosyltrasferase ( FUT8) gene (Mori et al., Biotechnol Bioeng88: 901-908, 2004), or coexpression of (3-1,4- N-acetylglucosaminyltransferase III and Golgi a-mannosidase II or a potent alpha- mannosidase I inhibitor, kifunensine (Ferrara et al., J Biol Chem281:5032-5036, 2006, Ferrara et al., Biotechnol Bioeng 93:851-861, 2006; Xhou et al., Biotechnol Bioeng 99:652-65, 2008).

PD-(L)1 axis Inhibitors

[00096] In one embodiment, the invention provides a generic concept for inhibiting the PD-(L)1 axis. In various embodiments, the composition comprises an inhibitor of one or more gene or protein in the PD-(L) 1 axis. In various embodiments, the present invention includes compositions and methods of decreasing the level or activity of one or more gene or protein in the PD-(L)1 axis. [00097] It will be understood by one skilled in the art, based upon the disclosure provided herein, that a decrease in the level or activity of one or more gene or protein in the PD- (L)l axis encompasses the decrease in the expression of the biomarker, including transcription, translation, or both. The skilled artisan will also appreciate, once armed with the teachings of the present invention, that a decrease in the level or activity of one or more gene or protein in the PD-(L)1 axis includes a decrease in the amount of polypeptide, a decrease in the amount of mRNA, a decrease in transcription, a decrease in translation, or a combination thereor; and it also includes decreasing any activity of one or more gene or protein in the PD-(L)1 axis as well.

[00098] Exemplary inhibitors of the PD-(L)1 axis include, but are not limited to, a small interfering RNA (siRNA), a microRNA, an antisense nucleic acid, a ribozyme, an expression vector encoding a transdominant negative mutant, an antibody, a fragment of an antibody, a fusion protein, an aptamer, a peptide and a small molecule.

[00099] One skilled in the art will appreciate, based on the disclosure provided herein, that one way to decrease the mRNA and/or protein levels of one or more PD-(L)1 axis protein in a cell is by reducing or inhibiting expression of the nucleic acid encoding the PD-(L)1 axis protein. Thus, the protein level of a PD-(L)1 axis protein in a cell can be decreased using a molecule or compound that inhibits or reduces gene expression such as, for example, siRNA, an antisense molecule or a ribozyme. However, the invention should not be limited to these examples.

[000100] In one embodiment, RNAi is used to decrease the level or activity of a PD-(L)1 axis protein. RNA interference (RNAi) is a phenomenon in which the introduction of double-stranded RNA (dsRNA) into a diverse range of organisms and cell types causes degradation of the complementary mRNA. In the cell, long dsRNAs are cleaved into short 21-25 nucleotide small interfering RNAs, or siRNAs, by a ribonuclease known as Dicer. The siRNAs subsequently assemble with protein components into an RNA-induced silencing complex (RISC), unwinding in the process. Activated RISC then binds to complementary transcript by base pairing interactions between the siRNA antisense strand and the mRNA. The bound mRNA is cleaved and sequence specific degradation of mRNA results in gene silencing. Chemical modification to siRNAs can aid in intravenous systemic delivery. Optimizing siRNAs involves consideration of overall G/C content, C/T content at the termini, Tm and the nucleotide content of the 3 ’ overhang. Therefore, the present invention also includes methods of decreasing levels of one or more PD-(L)1 axis protein using RNAi technology. [000101] In some embodiments, the invention includes an isolated nucleic acid encoding an inhibitor, such as a protein, an antibody, an siRNA or an antisense molecule operably linked to a nucleic acid comprising a promoter/regulatory sequence such that the nucleic acid is preferably capable of directing expression of the inhibitor encoded by the nucleic acid. Thus, the invention encompasses expression vectors and methods for the introduction of exogenous DNA into cells with concomitant expression of the exogenous DNA in the cells.

[000102] In order to assess the expression of the inhibitor, the expression vector to be introduced into a cell can also contain either a selectable marker gene or a reporter gene or both to facilitate identification and selection of expressing cells from the population of cells sought to be transfected or infected through viral vectors. In other embodiments, the selectable marker may be carried on a separate piece of DNA and used in a co -transfection procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers are known in the art and include, for example, antibiotic -resistance genes, such as neomycin [000103] When the inhibitor of the invention is a small molecule, a small molecule antagonist may be obtained using standard methods known to the skilled artisan. Such methods include chemical organic synthesis or biological means. Biological means include purification from a biological source, recombinant synthesis and in vitro translation systems, using methods well known in the art.

[000104] Combinatorial libraries of molecularly diverse chemical compounds potentially useful in treating a variety of diseases and conditions are well known in the art as are method of making the libraries. The method may use a variety of techniques well- known to the skilled artisan including solid phase synthesis, solution methods, parallel synthesis of single compounds, synthesis of chemical mixtures, rigid core structures, flexible linear sequences, deconvolution strategies, tagging techniques, and generating unbiased molecular landscapes for lead discovery vs. biased structures for lead development.

[000105] In a general method for small library synthesis, an activated core molecule is condensed with a number of building blocks, resulting in a combinatorial library of covalently linked, core-building block ensembles. The shape and rigidity of the core determines the orientation of the building blocks in shape space. The libraries can be biased by changing the core, linkage, or building blocks to target a characterized biological structure (“focused libraries”) or synthesized with less structural bias using flexible cores. [000106] In another aspect of the invention, one or more protein in the PD-(L)1 axis can be inhibited by way of inactivating and/or sequestering the protein(s). As such, inhibiting the effects of one or more protein in the PD-(L)1 axis can be accomplished by using a transdominant negative mutant.

[000107] In one embodiment, an antibody specific for one or more protein in the PD-(L)1 axis may be used. As will be understood by one skilled in the art, any antibody that can recognize and bind to an antigen of interest is useful in the present invention. Methods of making and using antibodies are well known in the art. For example, polyclonal antibodies useful in the present invention are generated by immunizing rabbits according to standard immunological techniques well-known in the art. Such techniques include immunizing an animal with a chimeric protein comprising a portion of another protein such as a maltose binding protein or glutathione (GSH) tag polypeptide portion, and/or a moiety such that the antigenic protein of interest is rendered immunogenic (e.g., an antigen of interest conjugated with keyhole limpet hemocyanin, KLH) and a portion comprising the respective antigenic protein amino acid residues. The chimeric proteins are produced by cloning the appropriate nucleic acids encoding the marker protein into a plasmid vector suitable for this purpose, such as but not limited to, pMAL-2 or pCMX. [000108] Exemplary anti-PD-(L)l axis antibodies include, but are not limited to, nivolumab (OPDIVO®), pembrolizumab (KEYTRUDA®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab or cetrelimab, or antibodies that bind PD-L1, such as PD-L1 antibodies are envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB- A317, TSR-042, and SHR-1210. In some embodiments, the anti-PD-(L)l axis antibodies are anti-PD-1 antibodies, such as antibodies specifically binding PD-1.

[000109] It is generally known in the art that each anti-PD-(L)l axis inhibitor antibody has its own unique efficacy and safety profile.

[000110] In some embodiments, the anti-PD-(L)l axis antibody comprises: a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26. In some embodiments, the anti-PD-(L)l axis antibody comprises a heavy chain variable domain (VH) of SEQ ID NO: 27 and a light chain variable domain (VL) of SEQ ID NO: 28. In some embodiments, the anti-PD-(L)l axis antibody comprises a heavy chain (HC) of SEQ ID NO: 29 and a light chain (LC) of SEQ ID NO: 30.

[000111] In some embodiments, the anti-PD-(L)! axis antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36. In some embodiments, the anti-PD-(L)! axis antibody comprises a heavy chain variable domain (VH) of SEQ ID NO: 37 and a light chain variable domain (VL) of SEQ ID NO: 38. In some embodiments, the anti-PD-(L)! axis antibody comprises a heavy chain (HC) of SEQ ID NO: 39 and a light chain (LC) of SEQ ID NO: 40.

[000112] However, the invention should not be construed as being limited solely to methods and compositions including these antibodies. Rather, the invention should be construed to include other antibodies, antibody fragments, or antibody mimetics, to PD- (L)l protein(s), or portions thereof.

[000113] One skilled in the art would appreciate, based upon the disclosure provided herein, that the antibody can specifically bind with any portion of the PD-(L)1 axis protein(s) and the full-length protein can be used to generate antibodies specific therefor. However, the present invention is not limited to using the full-length protein as an immunogen. Rather, the present invention includes using an immunogenic portion of the protein to produce an antibody that specifically binds with a specific antigen. That is, the invention includes immunizing an animal using an immunogenic portion, or antigenic determinant, of the antigen.

[000114] Once armed with the sequence of a specific antigen of interest and the detailed analysis localizing the various conserved and non-conserved domains of the protein, the skilled artisan would understand, based upon the disclosure provided herein, how to obtain antibodies specific for the various portions of the antigen using methods well-known in the art or to be developed.

[000115] The skilled artisan would appreciate, based upon the disclosure provided herein, that that present invention includes use of a single antibody recognizing a single antigenic epitope but that the invention is not limited to use of a single antibody. Instead, the invention encompasses use of at least one antibody where the antibodies can be directed to the same or different antigenic protein epitopes. [000116] The generation of polyclonal antibodies is accomplished by inoculating the desired animal with the antigen and isolating antibodies which specifically bind the antigen therefrom using standard antibody production methods.

[000117] Monoclonal antibodies directed against full length or peptide fragments of a protein or peptide may be prepared using any well-known monoclonal antibody preparation procedures. Quantities of the desired peptide may also be synthesized using chemical synthesis technology. Alternatively, DNA encoding the desired peptide may be cloned and expressed from an appropriate promoter sequence in cells suitable for the generation of large quantities of peptide. Monoclonal antibodies directed against the peptide are generated from mice immunized with the peptide using standard procedures as referenced herein.

[000118] The present invention also includes the use of humanized antibodies specifically reactive with epitopes of an antigen of interest. The humanized antibodies of the invention have a human framework and have one or more complementarity determining regions (CDRs) from an antibody, typically a mouse antibody, specifically reactive with an antigen of interest. When the antibody used in the invention is humanized, the antibody may be generated by expressing recombinant DNA segments encoding the heavy and light chain complementarity determining regions (CDRs) from a donor immunoglobulin capable of binding to a desired antigen, such as an epitope on an antigen of interest, attached to DNA segments encoding acceptor human framework regions. Generally speaking, the DNA segments will typically include an expression control DNA sequence operably linked to the humanized immunoglobulin coding sequences, including naturally -associated or heterologous promoter regions. The expression control sequences can be eukaryotic promoter systems in vectors capable of transforming or transfecting eukaryotic host cells or the expression control sequences can be prokaryotic promoter systems in vectors capable of transforming or transfecting prokaryotic host cells. Once the vector has been incorporated into the appropriate host, the host is maintained under conditions suitable for high level expression of the introduced nucleotide sequences and as desired.

[000119] The invention also includes antibody mimetics to functional equivalents of the antibodies described herein. Functional equivalents have binding characteristics comparable to those of the antibodies, and include, for example, hybridized and single chain antibodies, as well as fragments thereof.

[000120] Functional equivalents include polypeptides with amino acid sequences substantially the same as the amino acid sequence of the variable or hypervariable regions of the antibodies. “Substantially the same” amino acid sequence is defined herein as a sequence with at least 70%, 80%, 90%, 95%, or 99% identity to another amino acid sequence (or any integer in between 70 and 99), as determined by a sequence similarity search algorithm. Chimeric or other hybrid antibodies have constant regions derived substantially or exclusively from human antibody constant regions and variable regions derived substantially or exclusively from the sequence of the variable region of a monoclonal antibody from each stable hybridoma.

[000121] Single chain antibodies (scFv) or Fv fragments are polypeptides that consist of the variable region of the heavy chain of the antibody linked to the variable region of the light chain, with or without an interconnecting linker. Thus, the Fv comprises an antibody combining site. In one embodiment, the scFv can be fused to a half-life extending moiety, for example an Fc, using methods known to a skilled artisan.

[000122] Functional equivalents of the antibodies of the invention further include fragments of antibodies that have the same, or substantially the same, binding characteristics to those of the whole antibody. Such fragments may contain one or both Fab fragments or the F(ab')2 fragment. The antibody fragments contain all six complement determining regions of the whole antibody, although fragments containing fewer than all of such regions, such as three, four or five complement determining regions, are also functional. The functional equivalents are members of the IgG immunoglobulin class and subclasses thereof, but may be or may combine with any one of the following immunoglobulin classes: IgM, IgA, IgD, or IgE, and subclasses thereof. Heavy chains of various subclasses, such as the IgG subclasses, are responsible for different effector functions and thus, by choosing the desired heavy chain constant region, hybrid antibodies with desired effector function are produced. Exemplary constant regions are gamma 1 (IgGl), gamma 2 (IgG2), gamma 3 (IgG3), and gamma 4 (IgG4). The light chain constant region can be of the kappa or lambda type.

[000123] The immunoglobulins of the present invention can be monovalent, divalent or polyvalent. Monovalent immunoglobulins are dimers (HL) formed of a hybrid heavy chain associated through disulfide bridges with a hybrid light chain. Divalent immunoglobulins are tetramers (H2L2) formed of two dimers associated through at least one disulfide bridge. Additional Agents

[000124] In some embodiments, the combination therapy of the invention comprises a bispecific anti-EGFR/c-Met antibody and a PD-(L)1 axis inhibitor as described above in combination with one or more additional therapeutic agent.

[000125] In some embodiments, the combination therapy of the invention comprises one or more additional anti-cancer agent. In some embodiments, the additional anti-cancer agent is GM-CSF, a CD47 antagonist, an anti-CD47 antibody, an HD AC inhibitor, or a CD1 lb agonist.

[000126] Exemplary CD47 antagonists are CD47 ligand-Fc fusions, such as SIRPa- Fc fusions, such as TTI621 and ani-CD47 antibodies.

[000127] Exemplary anti-CD47 antibodies are Hu5F9-G4, TI-061, TTI-622, AO- 176, IBI-188, ALX-148, SRF-231, CC-90002 and anti-CD47 antibodies disclosed in Int. Pat. Publ. No. WO2016/081423.

[000128] Exemplary HD AC inhibitors are vorinostad, romidepsin, chidamide, panobinostat, belinostat, pracinostat, abexinostat, entinostat, vafidemstat, GSK-2879552, ricolinostat, iadademstat, domatinostat, resminostat, AZD-9468, nanatinostat, CG-200745, mocetinostat, INCB-59872, IMG-7289, tinostamustine, RDN-929, YM-753, HG-146, NBM-BMX, TAK-418, seclidemstat, CKD-504, CKD-506, CC-90011, KA-2507 and citarinostat.

[000129] Marketed antibodies may be purchased via authorized distributor or pharmacy. The amino acid sequences structures of the small molecules can be found from USAN and/or INN submissions by the companies of from CAS registry.

Methods of Treatment

[000130] It is well-known in the art that drug development is an unpredictable field.

The lack of predictability in the art is evidenced, for example, by health authority requirements (such as those of the Food and Drug Administration) to establish a safe and effective dosing regimen for each individual drug candidate in clinical trials. Over the last decade (2011-2020), only 7.9% of all developmental drug candidates achieved FDA approval from a Phase I clinical study. See Clinical Development Success Rates and Contributing Factors 2011-2020. The rate of success is even lower in oncology, such that only 5.3% of oncology drug candidates succeed.

[000131] In the field of oncology, even for a drug that already has an established dose in a particular indication, the Food and Drug Administration (FDA) recommends further clinical studies to identify an optimal dose for a new indication; otherwise, patients may be exposed to unreasonable and significant risk, among other potential deficiencies. See, e.g., Optimizing the Dosage of Human Prescription Drugs and Biological Products for the Treatment of Oncologic Diseases; Draft Guidance for Industry; January 2023). [000132] The disclosure provides a method of treating a subject having a solid tumor, comprising administering a therapeutically effective amount of an isolated bispecific epidermal growth factor receptor (EGFR)/hepatocyte growth factor receptor (c- Met) antibody to the subject in combination with an agent for inhibition of the PD-(L)1 axis in the subject.

[000133] The disclosure provides a method of remodeling a tumor microenvironment of a subject having a solid tumor, comprising administering a therapeutically effective amount of an isolated bispecific epidermal growth factor receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody to the subject in combination with an agent for inhibition of the PD-(L)1 axis in the subject. In some embodiments, the method comprises inhibiting glycolysis in the tumor microenvironment of a subject having a solid tumor, comprising administering a therapeutically effective amount of an isolated bispecific epidermal growth factor receptor (EGFR)/hepatocyte growth factor receptor (c- Met) antibody to the subject in combination with an agent for inhibition of the PD-(L)1 axis in the subject. In some embodiments, the method comprises reducing lactate production in the tumor microenvironment of a subject having a solid tumor, comprising administering a therapeutically effective amount of an isolated bispecific epidermal growth factor receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody to the subject in combination with an agent for inhibition of the PD-(L)1 axis in the subject.

[000134] In some embodiments, the method comprises inhibiting EGFR and MET signaling pathways in a tumor cell of a subject having a solid tumor, comprising administering a therapeutically effective amount of an isolated bispecific epidermal growth factor receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody to the subject in combination with an agent for inhibition of the PD-(L)1 axis in the subject. In some embodiments, the method comprises targeting of EGFR and MET expressing tumor cells for destruction by immune effector cells, such as natural killer cells and macrophages, through antibody -dependent cellular cytotoxicity (ADCC) and trogocytosis mechanisms, comprising administering a therapeutically effective amount of an isolated bispecific EGFR/c-Met antibody to the subject in combination with an agent for inhibition of the PD-(L)1 axis in the subject.

[000135] The disclosure provides a method of increasing the level of immune cell infiltration into a tumor or tumor microenvironment of a subject having a solid tumor, comprising administering a therapeutically effective amount of an isolated bispecific epidermal growth factor receptor (EGFR)/hepatocyte growth factor receptor (c-Met) antibody to the subject in combination with an agent for inhibition of the PD-(L)1 axis in the subject.

[000136] “Level’’ of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) may be qualitative (e.g., presence or absence) or quantitative (e.g., absolute cell numbers, relative numbers, percent (%) from a total cell count or % positive cells in a field). In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is above the mean value immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in a biological sample from a healthy subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 55th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 60th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 65th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 70th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 75th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 80th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 85th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 90th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 95th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject In some embodiments, the level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) is about the 100th percentile value of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) observed in the biological sample from subjects having a comparative cancer type and stage to the subject. In some embodiments, the biological sample is a blood sample. In some embodiments, the biological sample is a tumor tissue biopsy.

[000137] The level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) in the tumor microenvironment of subjects may also be compared relative to the levels of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) in a biological sample from healty subjects. The increased level of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) may for example be about 1.5-fold, about 2-fold, about 2.5-fold, about 3-fold, about 3.5-fold, about 4-fold, about 4. -fold, about 5-fold, about 5.5-fold, about 6-fold, about 6.5-fold, about 7-fold, about 7.5-fold, about 8-fold, about 8.5-fold, about 9-fold or about 10-fold higher when compared to the levels of immune cells (e.g., CD8+ T cells, central memory cytotoxic T cells) in the biological sample from healty subjects. In some embodiments, the biological sample is a blood sample. In some embodiments, the biological sample is a tissue sample.

[000138] The level of immune cells may be identified from for example tumor tissue biopsies obtained from subjects using immunohistochemistry, for example using 4- 1BB, CD45RA, PD-1, CD25, CD28, CD3, CD56, FoxP3, CD45, CD4, CCR7, CD8a and 7-AAD as markers for T cells, and evaluating percentage of area of positive staining and comparing to non-tumor tissue. The level of immune cells may be identified from for example tumor tissue biopsies obtained from subjects having the EGFR or c-Met expressing tumor using an immune gene signature.

[000139] Monocytes may be identified from blood samples from the subjects having the EGFR or c-Met expressing tumors using fluorescent cell sorting using, for example using CD40, CD45RA, CD80, CD86, HLA-DR, CDllc, CD14, CD68, CD45, CD 11b, CD 19, CD 123, CD 15 and 7-AAD as monocyte markers. [000140] In some embodiments, levels of FcyRI or FcyRIIIa may be used to predict patient response to the combination therapy of the invention by providing more immune cell interactions. For example, in some embodiments, the level of FcyRI or FcyRIIIa is above the mean value of the level of FcyRI or FcyRIIIa observed in a biological sample from a healthy subject or in a comparator control.

[000141] The level of FcyRI or FcyRIIIa may be measured using immunohistochemistry on tumor tissue samples (such as fresh frozen or paraffin embedded tumor tissue sections. The level of FcyRI or FcyRIIIa may be expressed as percent (%) of FcyRI or FcyRIIIa cells within a microscope field. The level of FcyRI or FcyRIIIa may also be measured at the gene expression level using RNA isolated from tumor tissue samples, either as part of an immune gene signature panel, or as individual genes.

[000142] In some embodiments, the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

[000143] In some embodiments, the cancer is resistant to treatment with PD-(L)1 axis inhibitors. In some embodiments, the cancer is resistant to treatment with PD-1 inhibitors. In some embodiments, the cancer is resistant to treatment with anti-PD-Ll antibodies. In some embodiments, the cancer is resistant to treatment with PD-L1 inhibitors.

[000144] In some embodiments, the cancer is an EGFR or c-Met expressing cancer. In some embodiments, the EGFR or c-Met expressing cancer is an epithelial cell cancer. In some embodiments, the EGFR or c-Met expressing cancer is breast cancer. In some embodiments, the EGFR or c-Met expressing cancer is ovarian cancer. In some embodiments, the EGFR or c-Met expressing cancer is lung cancer. In some embodiments, the EGFR or c-Met expressing cancer is non-small cell lung cancer (NSCLC). In some embodiments, the EGFR or c-Met expressing cancer is lung adenocarcinoma. In some embodiments, the EGFR or c-Met expressing cancer is small cell lung cancer. In some embodiments, the EGFR or c-Met expressing cancer is colorectal cancer. In some embodiments, the EGFR or c-Met expressing cancer is anal cancer. In some embodiments, the EGFR or c-Met expressing cancer is prostate cancer. In some embodiments, the EGFR or c-Met expressing cancer is kidney cancer. In some embodiments, the EGFR or c-Met expressing cancer is bladder cancer. In some embodiments, the EGFR or c-Met expressing cancer is head and neck cancer. In some embodiments, the EGFR or c-Met expressing cancer is pharynx cancer. In some embodiments, the EGFR or c-Met expressing cancer is cancer of the nose. In some embodiments, the EGFR or c-Met expressing cancer is pancreatic cancer. In some embodiments, the EGFR or c-Met expressing cancer is skin cancer. In some embodiments, the EGFR or c-Met expressing cancer is oral cancer. In some embodiments, the EGFR or c-Met expressing cancer is cancer of the tongue. In some embodiments, the EGFR or c-Met expressing cancer is esophageal cancer. In some embodiments, the EGFR or c-Met expressing cancer is vaginal cancer. In some embodiments, the EGFR or c-Met expressing cancer is cervical cancer. In some embodiments, the EGFR or c-Met expressing cancer is cancer of the spleen. In some embodiments, the EGFR or c-Met expressing cancer is testicular cancer. In some embodiments, the EGFR or c-Met expressing cancer is gastric cancer. In some embodiments, the EGFR or c-Met expressing cancer is cancer of the thymus. In some embodiments, the EGFR or c-Met expressing cancer is colon cancer. In some embodiments, the EGFR or c-Met expressing cancer is thyroid cancer. In some embodiments, the EGFR or c-Met expressing cancer is liver cancer. In some embodiments, the EGFR or c-Met expressing cancer is hepatocellular carcinoma (HCC). In some embodiments, the EGFR or c-Met expressing cancer is sporadic or hereditary papillary renal cell carcinoma (PRCC).

[000145] In some embodiments, NSCLC includes squamous cell carcinoma, adenocarcinoma, and large cell carcinoma. In some embodiments, cells of the NSCLC have an epithelial phenotype. In some embodiments, the NSCLC has acquired resistance to treatment with one or more EGFR inhibitors.

[000146] In some embodiments, the EGFR or c-Met expressing cancer is associated with a wild-type EGFR, an EGFR activating mutation, an EGFR gene amplification, increased levels of circulating HGF, a wild-type c-Met, a c-Met activating mutation, a c- Met gene amplification or a mutant KRAS.

[000147] Exemplary EGFR activating mutations that may be associated with cancer include point mutations, deletion mutations, insertion mutations, inversions or gene amplifications that lead to an increase in at least one biological activity of EGFR, such as elevated tyrosine kinase activity, formation of receptor homodimers and heterodimers, enhanced ligand binding etc. Mutations can be located in any portion of an EGFR gene or regulatory region associated with an EGFR gene and include mutations in exon 18, 19, 20 or 21 or mutations in the kinase domain. Other examples of EGFR activating mutations are known in the art (see e.g., U.S. Pat. Publ. No. US2005/0272083). Information about EGFR and other ErbB receptors including receptor homo- and hetero-dimers, receptor ligands, autophosphorylation sites, and signaling molecules involved in ErbB mediated signaling is known in the art (see e.g., Hynes and Lane, Nature Reviews Cancer 5: 341- 354, 2005).

[000148] In some embodiments, the EGFR activating mutation is L718Q, G719A, G719X (X being any amino acid), L861X (X being any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitution, deletion of E746-A750, deletion of R748-P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Vai and Ala (SVA) between S768 and V769, insertion of Asn and Ser (NS) between P772 and H773, insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, or one or more deletions or one or more insertions in EGFR exon 20.

[000149] Exemplary c-Met activating mutations include point mutations, deletion mutations, insertion mutations, inversions or gene amplifications that lead to an increase in at least one biological activity of a c-Met protein, such as elevated tyrosine kinase activity, formation of receptor homodimers and heterodimers, enhanced ligand binding etc. Mutations can be located in any portion of the c-Met gene or regulatory regions associated with the gene, such as mutations in the kinase domain of c-Met. Exemplary c-Met activating mutations are mutations at residue positions N375, V13, V923, R175, V136, L229, S323, R988, S1058/T1010 and E168. Methods for detecting EGFR and c-Met mutations or gene amplifications are well known.

[000150] In some embodiments, the mutant KRAS has a G12V, G12C or G12A substitution.

[000151] In NSCLC, specific mutations in the EGFR gene are associated with high response rates (70-80%) to EGFR tyrosine kinase inhibitors (EGFR-TKIs). A 5 amino acid deletion in exon 19 or the point mutation L858R in EGFR are associated with EGFR- TKI sensitivity (Nakata and Gotoh, Expert Opin Ther Targets 16 :771-781, 2012). These mutations result in a ligand-independent activation of the EGFR kinase activity.

Activating EGFR mutations occur in 10-30% of NSCLC patients and are significantly more common in East Asians, women, never smokers, and patients with adenocarcinoma histology (Janne and Johnson Clin Cancer Res 12(14 Suppl): 4416s-4420s, 2006). EGFR gene amplification is also strongly correlated with response after EGFR-TKI treatment (Cappuzzo et al., J Natl Cancer Inst 97:643-55, 2005). EGFR exon 20 insertions have been associated with EGFR TKI resistance.

[000152] Although the majority of NSCLC patients with EGFR mutations initially respond to EGFR TKI therapy, virtually all acquire resistance that prevents a durable response. 50-60% of patients acquire resistance due to a second-site point mutation in the kinase domain of EGFR (T790M). Nearly 60% of all tumors that become resistant to EGFR tyrosine kinase inhibitors increase c-Met expression, amplify the c-Met gene, or increase its only known ligand, HGF (Turke et al., Cancer Cell, 17:77-88, 2010).

[000153] In some embodiments, the subject is homozygous for phenylalanine at position 158 of CD 16 or heterozygous for valine and phenylalanine at position 158 of CD 16.

[000154] Subject homozygous for phenylalanine at position 158 of CD 16 has a

FcyRIIIa- 158F/F genotype. Subject heterozygous for valine and pheynylalanine at position 158 of CD16 has a FcyRIIIa- 158F/V genotype. CD16 is also known as the Fc gamma receptor Illa (FcyRIIIa) or the low affinity immunoglobulin gamma Fc region receptor III-A isoform. Valine/phenylalanine (V/F) polymorphism at FcyRIIIa protein residue position 158 has been shown to affect FcyRIIIa affinity to human IgG. Receptor with FcyRIIIa-158F/F or FcyRIIIa-158F/V polymorphisms demonstrates reduced Fc engagement and therefore reduced ADCC when compared to the FcyRIIIa- 158 V/V. The lack of or low amount of fucose on human N-linked oligosaccharides improves the ability of the antibodies to induce ADCC due to improved binding of the antibodies to human FcyRIIIa (CD16) (Shields et al., J Biol Chem 277:26733-40, 2002).

[000155] In some embodiments, the subject has a newly diagnosed EGFR or c-Met expressing cancer. In some embodiments, the subject having the newly diagnosed EGFR or c-Met expressing cancer has one or more EGFR exon 20 mutation. Exon 20 mutations (insertion of one or more amino acids are generally resistant to EGFR tyrosine kinase inhibitors (TKI) (see. e.g. Int. Pat. Publ. No. WO2018/094225).

[000156] In some embodiments, the subject is resistant or has acquired resistance to treatment with a prior anti-cancer therapy.

[000157] In some embodiments, the prior anti-cancer therapy is chemotherapy, a targeted anti-cancer therapy or a kinase inhibitor.

[000158] In some embodiments, the kinase inhibitor is an inhibitor of EGFR, c-Met,

HER2, HER3, HER4, VEGFR or AXL. [000159] In some embodiments, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib. [000160] In some embodiments, the subject is resistant or has acquired resistance to an EGFR inhibitor. Exemplary EGFR inhibitors for which cancer may acquire resistance are anti-EGFR antibodies cetuximab (ERBITUX®), pantinumumab (VECTIBIX®), matuzumab, nimotuzumab, small molecule EGFR inhibitors erlotinib (TARCEVA® ), gefitinib (IRESSA®), EKB-569 (pelitinib, irreversible EGFR TKI), pan-ErbB and other receptor tyrosine kinase inhibitors, lapatinib (EGFR and HER2 inhibitor), pelitinib (EGFR and HER2 inhibitor), vandetanib (ZD6474, ZACTIMA™, EGFR, VEGFR2 and RET TKI), PF00299804 (dacomitinib, irreversible pan-ErbB TKI) , CI-1033 (irreversible pan- erbB TKI), afatinib (BIBW2992, irreversible pan-ErbB TKI), AV-412 (dual EGFR and ErbB2 inhibitor), EXEL-7647 (EGFR, ErbB2, GEVGR and EphB4 inhibitor), CO-1686 (irreversible mutant-selective EGFR TKI), AZD9291 (irreversible mutant-selective EGFR TKI), and HKI-272 (neratinib, irreversible EGFR/ErbB2 inhibitor).

[000161] Various qualitative and/or quantitative methods may be used to determine if a subject is resistant, has developed or is susceptible to developing a resistance to treatment with an anti-cancer therapy. Symptoms that may be associated with resistance to an anti-cancer therapy include a decline or plateau of the well-being of the patient, an increase in the size of a tumor, arrested or slowed decline in growth of a tumor, and/or the spread of cancerous cells in the body from one location to other organs, tissues or cells. Re-establishment or worsening of various symptoms associated with cancer may also be an indication that a subject has developed or is susceptible to developing resistance to an anti-cancer therapy, such as anorexia, cognitive dysfunction, depression, dyspnea, fatigue, hormonal disturbances, neutropenia, pain, peripheral neuropathy, and sexual dysfunction. The symptoms associated with cancer may vary according to the type of cancer. For example, symptoms associated with cervical cancer may include abnormal bleeding, unusual heavy vaginal discharge, pelvic pain that is not related to the normal menstrual cycle, bladder pain or pain during urination, and bleeding between regular menstrual periods, after sexual intercourse, douching, or pelvic exam. Symptoms associated with lung cancer may include persistent cough, coughing up blood, shortness of breath, wheezing chest pain, loss of appetite, losing weight without trying and fatigue.

Symptoms for liver cancer may include loss of appetite and weight, abdominal pain, especially in the upper right part of abdomen that may extend into the back and shoulder, nausea and vomiting, general weakness and fatigue, an enlarged liver, abdominal swelling (ascites), and a yellow discoloration of the skin and the whites of eyes (jaundice). One skilled in oncology may readily identify symptoms associated with a particular cancer type.

[000162] In some embodiments, the subject is further administered one or more additional anti-cancer therapy.

[000163] In some embodiments, the additional anti-cancer therapy is chemotherapy, a targeted anti-cancer therapy or a kinase inhibitor.

[000164] In some embodiments, the kinase inhibitor is an inhibitor of EGFR, c-Met,

HER2, HER3, HER4, VEGFR or AXL. In some embodiments, the kinase inhibitor is an inhibitor of EGFR. In some embodiments, the kinase inhibitor is an inhibitor of c-Met. In some embodiments, the kinase inhibitor is an inhibitor of HER2. In some embodiments, the kinase inhibitor is an inhibitor of HER3. In some embodiments, the kinase inhibitor is an inhibitor of HER4. In some embodiments, the kinase inhibitor is an inhibitor of VEGFR. In some embodiments, the kinase inhibitor is an inhibitor of or AXL.

[000165] In some embodiments, the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib. [000166] In some embodiments, the kinase inhibitor is erlotinib. In some embodiments, the kinase inhibitor is gefitinib. In some embodiments, the kinase inhibitor is lapatinib. In some embodiments, the kinase inhibitor is vandetanib. In some embodiments, the kinase inhibitor is afatinib. In some embodiments, the kinase inhibitor is osimertinib. In some embodiments, the kinase inhibitor is lazertinib. In some embodiments, the kinase inhibitor is poziotinib. In some embodiments, the kinase inhibitor is criotinib. In some embodiments, the kinase inhibitor is cabozantinib. In some embodiments, the kinase inhibitor is capmatinib. In some embodiments, the kinase inhibitor is axitinib. In some embodiments, the kinase inhibitor is lenvatinib. In some embodiments, the kinase inhibitor is nintedanib. In some embodiments, the kinase inhibitor is regorafenib. In some embodiments, the kinase inhibitor is pazopanib. In some embodiments, the kinase inhibitor is sorafenib. In some embodiments, the kinase inhibitor is sunitinib.

[000167] Anti-cancer therapies that may be administered in combination with the combination of the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitory agent in the methods of the disclosure include any one or more of the chemotherapeutic drugs or other anti-cancer therapeutics known to those of skill in the art.

Chemotherapeutic agents are chemical compounds useful in the treatment of cancer and include growth inhibitory agents or other cytotoxic agents and include alkylating agents, anti-metabolites, anti-microtubule inhibitors, topoisomerase inhibitors, receptor tyrosine kinase inhibitors, angiogenesis inhibitors and the like. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethylenethiophosphaoramide and trimethylolomelamine; nitrogen mustards such as chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-FU; folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6- azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirambicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; members of taxoid or taxane family, such as paclitaxel (TAXOL®docetaxel (TAXOTERE®) and analogues thereof; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylomithine (DMFO); retinoic acid; esperamicins; capecitabine; inhibitors of receptor tyrosine kinases and/or angiogenesis, including sorafenib (NEXAVAR® ), sunitinib (SUTENT® ), pazopanib (VOTRIENT™), toceranib (PALLADIA™), vandetanib (ZACTIMA™), cediranib (RECENTIN®), regorafenib (BAY 73-4506), axitinib (AG013736), lestaurtinib (CEP-701), erlotinib (TARCEVA®), gefitinib (IRESSA®), afatinib (BIBW 2992), lapatinib (TYKERB®), neratinib (HKI-272), and the like, and pharmaceutically acceptable salts, acids or derivatives of any of the above. Also included in this definition are anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as anti-estrogens including for example tamoxifen, raloxifene, aromatase inhibiting 4(5)-imidazoles, 4- hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, and toremifene (FARESTON®); and anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above. Other conventional cytotoxic chemical compounds as those disclosed in Wiemann et al., 1985, in Medical Oncology (Calabresi et aL, eds.), Chapter 10, McMillan Publishing, are also applicable to the methods of the present invention.

Administration

[000168] The bispecific anti -EGFR/c -Met antibody and the PD-(L)1 axis inhibitory agent may be administered to the subject together in a mixture, concurrently as single agents or sequentially as single agents in any order.

[000169] In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered prior to administration of the PD-(L)1 axis inhibitory agent.

[000170] In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered after administration of the PD-(L)1 axis inhibitory agent. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered to a subject having received a prior administration of a PD-(L)1 axis inhibitory agent. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered to a subject having received a prior administration of an anti-PD-(L)l axis antibody. In some embodiments, the prior administration of an anti-PD-(L)l axis antibody was provided as a prior administered cancer therapy or in combination with a prior administered cancer therapy.

[000171] In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered simultaneously with administration of the PD-(L)1 axis inhibitory agent. [000172] In some embodiments, the combination of the bispecific anti-EGFR/c-Met antibody and PD-(L)1 axis inhibitory agent is administered prior to administration of one or more additional anti-cancer agent.

[000173] In some embodiments, the combination of the bispecific anti-EGFR/c-Met antibody and PD-(L)1 axis inhibitory agent is administered following administration of one or more additional anti-cancer agent.

[000174] In some embodiments, the combination of the bispecific anti-EGFR/c-Met antibody and PD-(L)1 axis inhibitory agent is administered simultaneously with administration of one or more additional anti-cancer agent.

[000175] The length of time between administrations of the bispecific anti-EGFR/c- Met antibody and the PD-(L)1 axis inhibitory agent or one or more additional anti-cancer agent may be a few minutes, such as about 1, 2, 5, 10, 30 or 60 minutes or several hours, such as about 2, 4, 6, 10, 12, 24 or 36 hours, or such as about 2, 4, 7, 14, 21, 28, 35, 42, 49, 56 days or more.

[000176] In some embodiments, a bispecific anti-EGFR/c-Met antibody is administered about 1, 2, 5, 10, 30 or 60 minutes or several hours, such as about 2, 4, 6, 10, 12, 24 or 36 hours, or such as about 2, 4, 7, 14, 21, 28, 35, 42, 49, 56 days or more following administration of a PD-(L)1 axis inhibitory agent. Therefore, in some embodiments, a bispecific anti-EGFR/c-Met antibody is administered to a subject who has previously been administered a PD-(L)1 axis inhibitory agent.

[000177] In some embodiments, a bispecific anti-EGFR/c-Met antibody is administered about 1, 2, 5, 10, 30 or 60 minutes or several hours, such as about 2, 4, 6, 10, 12, 24 or 36 hours, or such as about 2, 4, 7, 14, 21, 28, 35, 42, 49, 56 days or more prior to administration of a PD-(L)1 axis inhibitory agent. Therefore, in some embodiments, a PD- (L) 1 axis inhibitory agent is administered to a subject who has previously been administered a bispecific anti-EGFR/c-Met antibody.

[000178] One or more of the bispecific anti-EGFR/c-Met antibody, the PD-(L)1 axis inhibitory agent or an additional anti-cancer agent may be administered in a pharmaceutically acceptable carrier. "Carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the antibody of the invention is administered. Such vehicles may be liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. For example, 0.4% saline and 0.3% glycine may be used to formulate the bispecific anti- EGFR/c-Met antibody. These solutions are sterile and generally free of particulate matter. They may be sterilized by conventional, well-known sterilization techniques (e.g., filtration). For solid oral preparations, such as powders capsules and tablets, suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like. Solid oral preparations may also be coated with substances such as sugars or be enteric-coated to modulate major site of absorption. For parenteral administration, the carrier may comprise sterile water and other excipients may be added to increase solubility or preservation. Injectable suspensions or solutions may also be prepared utilizing aqueous carriers along with appropriate additives. Suitable vehicles and formulations, inclusive of other human proteins, e.g., human serum albumin, are described, for example, in e.g. Remington: The Science and Practice of Pharmacy, 21st Edition, Troy, D.B. ed., Lipincott Williams and Wilkins, Philadelphia, PA 2006, Part 5, Pharmaceutical Manufacturing pp 691-1092, See especially pp. 958-989.

[000179] The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, stabilizing, thickening, lubricating and coloring agents, etc. The concentration of the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitory agent in the pharmaceutical formulation may vary, from less than about 0.5%, usually to at least about 1% to as much as 15%, 20%, 30%, 40% or 50% by weight and may be selected primarily based on required dose, fluid volumes, viscosities, etc., according to the particular mode of administration selected. Pharmaceutical compositions comprising solid forms may contain about 0.1 mg to about 2000 mg, such as about 1 mg, about 5 mg, about 10 mg, about 25 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 300 mg, about 500 mg about 600 mg or about 1000 mg of active ingredient.

[000180] In some embodiments, amivantamab is administered intravenously. In some embodiments, amivantamab is administered subcutaneously.

[000181] In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 350 mg and about 3000 mg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of between about 350 mg and about 4650 mg.

[000182] In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 350 mg, about 700 mg, about 1050 mg, about 1400 mg, about 1600 mg, about 1750 mg, about 2100 mg, about 2240 mg, about 2,400 mg, about 3,360 mg, about 3,200 mg, about 4,320 mg, about 3,520 mg, or about 4,640 mg.. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 350 mg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 700 mg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 1050 mg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 1400 mg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 1600 mg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 1750 mg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 2400 mg. In some embodiments, the antibody is administered at a dose of about 3,360 mg. In some embodiments, the antibody is administered at a dose of about 3,200 mg. In some embodiments, the antibody is administered at a dose of about 4,320 mg. In some embodiments, the antibody is administered at a dose of about 3,520 mg. In some embodiments, the antibody is administered at a dose of about 4,640 mg. In some embodiments, the bispecific anti- EGFR/c-Met antibody is administered at a dose of about 2240 mg or about 3000 mg. [000183] In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose of about 700 mg for body weight <80 kg and 1050 mg for body weight > 80 kg. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered on a 28-day cycle. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered once weekly for the first 4 weeks (cycle 1). In some embodiments, the first dose is split over the first 2 days. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered every 2 weeks (Q2W) starting cycle 2 onwards. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose greater than 700 mg for body weight <80 kg starting cycle 2 onwards. In some embodiments, the bispecific anti-EGFR/c-Met antibody is administered at a dose greater than 1050 mg for body weight > 80 kg starting cycle 2 onwards.

[000184] In some embodiments, cetrelimab is administered intravenously. In some embodiments, cetrelimab is administered at a dose of about 240 mg. In some embodiments, cetrelimab is administered once every 2 weeks (Q2W). In some embodiments, cetrelimab is administered at a dose of about 240 mg once every 2 weeks (Q2W). In some embodiments, cetrelimab is administered at a dose of about 480 mg. In some embodiments, cetrelimab is administered once every 4 weeks (Q4W). In some embodiments, cetrelimab is administered at a dose of about 480 mg once every 4 weeks (Q4W). In some embodiments, the first dose of cetrelimab is administered on day 2 of cycle 1. In some embodiments, cetrelimab is administered on a 28-day cycle. Bispecific anti-EGFR/c-Met antibody Sequences

[000185] An exemplary anti-EGFR/c-Met antibody that can be used in the methods of the disclosures is amivantamab. Amivantamab is characterized by following amino acid sequences:

EGFR binding arm

>SEQ ID NO: 1 (HCDR1, EGFR binding arm)

TYGMH

>SEQ ID NO: 2 (HCDR2, EGFR binding arm) VIWDDGSYKYYGDSVKG

>SEQ ID NO: 3 (HCDR3, EGFR binding arm)

DGITMVRGVMKDYFDY

>SEQ ID NO: 4 (LCDRl, EGFR binding arm)

RASQDISSALV

>SEQ ID NO: 5 (LCDR2, EGFR binding arm) DASSLES

>SEQ ID NO: 6 (LCDR3, EGFR binding arm)

QQFNSYPLT

>SEQ ID NO: 7 (HCDR1, c-Met binding arm) SYGIS

>SEQ ID NO: 8 (HCDR2, c-Met binding arm)

WISAYNGYTNYAQKLQG

>SEQ ID NO:9 (HCDR3, c-Met binding arm) DLRGTNYFDY

>SEQ ID NO: 10 (LCDRl, c-Met binding arm)

RASQGISNWLA >SEQ ID NO: 11 (LCDR2, c-Met binding arm)

AASSLLS

>SEQ ID NO: 12 (LCDR3, c-Met binding arm)

QQANSFPIT

>SEQ ID NO: 13 (VH, EGFR binding arm)

QVQLVESGGGWQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWDDG SYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGVMKD YFDYWGQGTLVTVSS

>SEQ ID NO: 14 (VL, EGFR binding arm)

AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESGVP

SRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK

>SEQ ID NO: 15 (VH, c-Met binding arm)

QVQLVQSGAEVKKPGASVKVSCETSGYTFTSYGISWVRQAPGHGLEWMGWISAYN GYTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLRGTNYFDYWG QGTLVTVSS

>SEQ ID NO: 16 (VL, c-Met binding arm)

DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWFQHKPGKAPKLLIYAASSLLSGV

PSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPITFGQGTRLEIK

>SEQ ID NO: 17 HC1

QVQLVESGGGWQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEWVAVIWD

DGSYKYYGDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGITMVRGV MKDYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCWVDV

SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVE WESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGK >SEQ ID NO: 18 LC1

AIQLTQSPSSLSASVGDRVTITCRASQDISSALVWYQQKPGKAPKLLIYDASSLESG VPSRFSGSESGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKRTVAAPS VFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

>SEQ ID NO: 19 HC2

QVQLVQSGAEVKKPGASVKVSCETSGYTFTSYGISWVRQAPGHGLEWMGWISAY NGYTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLRGTNYFD YWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKPSNTKVDKRVE

PKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVFSCSVMHEALHNH

YTQKSLSLSPGK

>SEQ ID NO: 20 LC2

DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWFQHKPGKAPKLLIYAASS LLSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPITFGQGTRLEIKRTVAAP SVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

PD-(L)1 axis inhibitory antibody sequences

[000186] An exemplary anti-PD-(L)l axis antibody that can be used in the methods of the disclosures is Pembrolizumab. Pembrolizumab is characterized by following amino acid sequences:

>SEQ ID NO: 21 (HCDR1)

NYYMY

>SEQ ID NO: 22 (HCDR2)

GINPSNGGTNFNEKFKN

>SEQ ID NO: 23 (HCDR3)

RDYRFDMGFDY >SEQ ID NO: 24 (LCDR1)

RASKGVSTSGYSYLH

>SEQ ID NO: 25 (LCDR2)

LASYLES

>SEQ ID NO: 26 (LCDR3)

QHSRDLPLT

>SEQ ID NO: 27 (VH)

QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGIN PSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMG FDYWGQGTTVTVSS

>SEQ ID NO: 28 (VL)

EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLAS

YLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIK

>SEQ ID NO: 29 (HC)

QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGIN PSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMG FDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDK

RVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEV QFNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNK GLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYT

QKSLSLSLGK

>SEQ ID NO: 30 (LC)

EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLAS YLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRT VAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC [000187] An exemplary anti-PD-(L)l axis antibody that can be used in the methods of the disclosures is cetrelimab. Cetrelimab is characterized by following amino acid sequences:

SEQ ID NO: 31 (HCDR1)

SYAIS

SEQ ID NO: 32 (HCDR2)

GIIPIFDTANYAQKFQG

SEQ ID NO: 33 (HCDR3)

PGLAAAYDTGSLDY

SEQ ID NO: 34 (LCDR1)

RASQSVRSYLA

SEQ ID NO: 35 (LCDR2)

DASNRAT

SEQ ID NO: 36 (LCDR3)

QQRNYWPLT

SEQ ID NO: 37 (VH)

QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIF

DTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARPGLAAAYDTGSL

DYWGQGTLVTVSS

SEQ ID NO: 38 (VL)

EIVLTQSPATLSLSPGERATLSCRASQSVRSYLAWYQQKPGQAPRLLIYDASNRAT

GIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRNYWPLTFGQGTKVEIK

SEQ ID NO: 39 (HC)

QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIF

DTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARPGLAAAYDTGSL

DYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTKTYTCNVDHKPSNTKVDKR VESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQ FNWYVDGVEVHNAKTKPREEQFNSTYRWSVLTVLHQDWLNGKEYKCKVSNKG LPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQ KSLSLSLGK

SEQ ID NO: 40 (LC)

EIVLTQSPATLSLSPGERATLSCRASQSVRSYLAWYQQKPGQAPRLLIYDASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRNYWPLTFGQGTKVEIKRTVAA PSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

EMBODIMENTS

1. A method of treating a solid tumor or ameliorating cancer progression in a subject in need thereof, the method comprising administering to the subject:

(a) a PD-(L)1 axis inhibitor; and

(b) a bispecific anti-EGFR/c-Met antibody.

2. The method of embodiment 1, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

3. The method of embodiment 2, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. 4. The method of embodiment 2, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

5. The method of embodiment 4, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

5a. The method of any of the embodiments 1-5, wherein the bispecific anti- EGFR/c-Met antibody is amivantamab.

6. The method of embodiment 1, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

6a. The method of embodiment 1, wherein the PD-(L)1 axis inhibitor is an anti- PD-1 antibody.

7. The method of embodiment 6, wherein the inhibitory antibody is selected from the group consisting of cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

8. The method of embodiment 7, wherein the Pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26.

8a. The method of embodiment 7, wherein the Pembrolizumab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 27, and a light chain variable region (VL) of SEQ ID NO: 28.

9. The method of embodiment 7, wherein the Pembrolizumab antibody comprises a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO: 30.

10. The method of embodiment 7, wherein the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36. 10a. The method of embodiment 7, wherein the cetrelimab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 37, and a light chain variable region (VL) of SEQ ID NO: 38.

11. The method of embodiment 7, wherein the cetrelimab antibody comprises a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

12. The method of embodiment 1, wherein the cancer is an EGFR, c-Met or EGFR and c-Met expressing cancer associated with a wild-type EGFR, an EGFR activating mutation, an EGFR gene amplification, increased levels of circulating HGF, a wild-type c-Met, a c-Met activating mutation, a c-Met gene amplification or a mutant KRAS, or any combination thereof.

13. The method of embodiment 12, wherein the EGFR activating mutation comprises L718Q, G719A, G719X (X being any amino acid), L861X (X being any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitution, deletion of E746-A750, deletion of R748-P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Vai and Ala (SVA) between S768 and V769, insertion of Asn and Ser (NS) between P772 and H773, insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof.

14. The method of embodiment 13, wherein the mutant KRAS comprises a G12V, G12C or G12A substitution.

15. The method of any one of embodiments 1-14, wherein the method inceases immune cell infiltration into a solid tumor, further wherein the immune cells are selected from the group consisting of T cells, B cells, and natural killer cells.

16. The method of embodiment 15, wherein the T cells comprise CD8+ T cells and CD4+ T cells.

17. The method of embodiment 16, wherein the T cells comprise CD8+ T cells.

18. The method of any one of embodiments 1-17, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

19. The method of any one of embodiments 1-18, wherein the subject is suspected to have or has an EGFR, c-Met or EGFR and c-Met expressing cancer.

20. The method of embodiment 1, wherein the subject is resistant or has acquired resistance to treatment with a prior anti-cancer therapy. 21. The method of embodiment 20, wherein the prior anti-cancer therapy is a kinase inhibitor.

22. The method of embodiment 21, wherein the kinase inhibitor is an inhibitor of EGFR, c-Met, HER2, HER3, HER4, VEGFR or AXL.

23. The method of embodiment 22, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

24. The method of any one of embodiments 1-23, wherein the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

25. The method of any one of embodiments 1-24, wherein the cancer is resistant to treatment with PD-(L)1 axis inhibitors.

26. The method of any one of embodiments 1-25, wherein the cancer is selected from the group consisting of head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), or non-small cell lung cancer (NSCLC).

27. The method of any one of embodiments 1-26, further comprising administering one or more anti-cancer therapies to the subject.

28. The method of embodiment 27, wherein the one or more anti-cancer therapies comprise a kinase inhibitor.

29. The method of embodiment 28, wherein the kinase inhibitor is an inhibitor of EGFR, c- Met, HER2, HER3, HER4, VEGFR or AXL.

30. The method of embodiment 29, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

31. The method of embodiment 20, wherein the prior anti-cancer therapy is chemotherapy. 32. The method of embodiment 20, wherein the prior anti-cancer therapy is a targeted anti-cancer therapy.

33. The method of embodiment 27, wherein the one or more anti-cancer therapies comprise chemotherapy.

34. The method of embodiment 27, wherein the one or more anti-cancer therapies comprise a targeted anti-cancer therapy.

35. The method of any one of embodiments 1-34, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L) 1 axis inhibitor are administered essentially concurrently to at two different injection sites.

36. The method of any one of embodiments 1-34, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are combined prior to administration and administered simultaneously.

37. The method of any one of embodiments 1-34, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L) 1 axis inhibitor are administered sequentially over the course of one or more days as part of a multi-step treatment regimen.

38. A method of enhancing immune cell infiltration into a solid tumor in a subject in need thereof, the method comprising administering to the subject:

(a) a PD-(L)1 axis inhibitor; and

(b) a bispecific anti-EGFR/c-Met antibody.

39. The method of embodiment 38, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

40. The method of embodiment 39, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16. 41. The method of embodiment 39, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

42. The method of embodiment 41, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

43. The method of embodiment 38, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

43a. The method of embodiment 38, wherein the PD-(L)1 axis inhibitor is an anti- PD-1 antibody.

44. The method of embodiment 43, wherein the inhibitory antibody is selected from the group consisting of pembrolizumab (KEYTRUDA®), cetrelimab, nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

45. The method of embodiment 44, wherein the Pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26.

45a. The method of embodiment 44, wherein the Pembrolizumab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 27, and a light chain variable region (VL) of SEQ ID NO: 28.

46. The method of embodiment 44, wherein the Pembrolizumab antibody comprises a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO:

30.

47. The method of embodiment 44, wherein the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO:

31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36.

47a. The method of embodiment 44, wherein the cetrelimab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 37, and a light chain variable region (VL) of SEQ ID NO: 38. 48. The method of embodiment 44, wherein the cetrelimab antibody comprises a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

49. The method of embodiment 38, wherein the cancer is an EGFR, c-Met or EGFR and c-Met expressing cancer associated with a wild-type EGFR, an EGFR activating mutation, an EGFR gene amplification, increased levels of circulating HGF, a wild-type c-Met, a c-Met activating mutation, a c-Met gene amplification or a mutant KRAS, or any combination thereof.

50. The method of embodiment 49, wherein the EGFR activating mutation comprises L718Q, G719A, G719X (X being any amino acid), L861X (X being any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitution, deletion of E746-A750, deletion of R748-P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Vai and Ala (SVA) between S768 and V769, insertion of Asn and Ser (NS) between P772 and H773, insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof.

51. The method of embodiment 50, wherein the mutant KRAS comprises a G12V, G12C or G12A substitution.

52. The method of any one of embodiments 38-51, wherein the immune cells are selected from the group consisting of T cells, B cells, and natural killer cells.

53. The method of embodiment 52, wherein the T cells comprise CD8+ T cells and CD4+ T cells.

54. The method of embodiment 53, wherein the T cells comprise CD8+ T cells.

55. The method of any one of embodiments 38-54, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

56. The method of any one of embodiments 38-55, wherein the subject is suspected to have or has an EGFR, c-Met or EGFR and c-Met expressing cancer.

57. The method of embodiment 38, wherein the subject is resistant or has acquired resistance to treatment with a prior anti-cancer therapy. 58. The method of embodiment 57, wherein the prior anti-cancer therapy is a kinase inhibitor.

59. The method of embodiment 58, wherein the kinase inhibitor is an inhibitor of EGFR, c-Met, HER2, HER3, HER4, VEGFR or AXL.

60. The method of embodiment 59, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

61. The method of any one of embodiments 38-60, wherein the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

62. The method of any one of embodiments 38-61, wherein the cancer is resistant to treatment with PD-(L)1 axis inhibitors.

63. The method of any one of embodiments 38-62, wherein the cancer is selected from the group consisting of head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), or non-small cell lung cancer (NSCLC).

64. The method of any one of embodiments 38-63, further comprising administering one or more anti-cancer therapies to the subject.

65. The method of embodiment 64, wherein the one or more anti-cancer therapies comprise a kinase inhibitor.

66. The method of embodiment 65, wherein the kinase inhibitor is an inhibitor of EGFR, c- Met, HER2, HER3, HER4, VEGFR or AXL.

67. The method of embodiment 66, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

68. The method of embodiment 57, wherein the prior anti-cancer therapy is chemotherapy. 69. The method of embodiment 57, wherein the prior anti-cancer therapy is a targeted anti-cancer therapy.

70. The method of embodiment 64, wherein the one or more anti-cancer therapies comprise chemotherapy.

71. The method of embodiment 64, wherein the one or more anti-cancer therapies comprise a targeted anti-cancer therapy.

72. The method of any one of embodiments 38-71, wherein the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered essentially concurrently to at two different injection sites.

73. The method of any one of embodiments 38-71, wherein the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are combined prior to administration and administered simultaneously.

74. The method of any one of embodiments 38-71, wherein the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered sequentially over the course of one or more days as part of a multi-step treatment regimen.

75. A method of reducing glycolysis or lactic acid production in the tumor microenvironment in a subject in need thereof, the method comprising administering to the subject:

(a) a PD-(L)1 axis inhibitor; and

(b) a bispecific anti-EGFR/c-Met antibody.

76. The method of embodiment 75, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

77. The method of embodiment 76, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and (b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

78. The method of embodiment 76, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

79. The method of embodiment 78, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

80. The method of embodiment 75, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

80a. The method of embodiment 75, wherein the PD-(L)1 axis inhibitor is an anti- PD-1 antibody.

81. The method of embodiment 80, wherein the inhibitory antibody is selected from the group consisting of pembrolizumab (KEYTRUDA®), cetrelimab, nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

82. The method of embodiment 81, wherein the Pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26.

82a. The method of embodiment 81, wherein the Pembrolizumab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 27, and a light chain variable region (VL) of SEQ ID NO: 28.

83. The method of embodiment 81, wherein the Pembrolizumab antibody comprises a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO:

30.

84. The method of embodiment 81, wherein the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO:

31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36. 84a. The method of embodiment 81, wherein the cetrelimab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 37, and a light chain variable region (VL) of SEQ ID NO: 38.

85. The method of embodiment 81, wherein the cetrelimab antibody comprises a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

86. The method of embodiment 75, wherein the cancer is an EGFR, c-Met or EGFR and c-Met expressing cancer associated with a wild-type EGFR, an EGFR activating mutation, an EGFR gene amplification, increased levels of circulating HGF, a wild-type c-Met, a c-Met activating mutation, a c-Met gene amplification or a mutant KRAS, or any combination thereof.

87. The method of embodiment 86, wherein the EGFR activating mutation comprises L718Q, G719A, G719X (X being any amino acid), L861X (X being any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitution, deletion of E746-A750, deletion of R748-P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Vai and Ala (SVA) between S768 and V769, insertion of Asn and Ser (NS) between P772 and H773, insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof.

88. The method of embodiment 87, wherein the mutant KRAS comprises a G12V, G12C or G12A substitution.

89. The method of any one of embodiments 75-88, wherein the method inceases immune cell infiltration into a solid tumor, further wherein the immune cells are selected from the group consisting of T cells, B cells, and natural killer cells.

90. The method of embodiment 89, wherein the T cells comprise CD8+ T cells and CD4+ T cells.

91. The method of embodiment 90, wherein the T cells comprise CD8+ T cells.

92. The method of any one of embodiments 75-91, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

93. The method of any one of embodiments 75-92, wherein the subject is suspected to have or has an EGFR, c-Met or EGFR and c-Met expressing cancer. 94. The method of embodiment 75, wherein the subject is resistant or has acquired resistance to treatment with a prior anti-cancer therapy.

95. The method of embodiment 94, wherein the prior anti-cancer therapy is a kinase inhibitor.

96. The method of embodiment 95, wherein the kinase inhibitor is an inhibitor of EGFR, c-Met, HER2, HER3, HER4, VEGFR or AXL.

97. The method of embodiment 96, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

98. The method of any one of embodiments 75-97, wherein the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

99. The method of any one of embodiments 75-98, wherein the cancer is resistant to treatment with PD-(L)1 axis inhibitors.

100. The method of any one of embodiments 75-99, wherein the cancer is selected from the group consisting of head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), or non-small cell lung cancer (NSCLC).

101. The method of any one of embodiments 75-100, further comprising administering one or more anti-cancer therapies to the subject.

102. The method of embodiment 101, wherein the one or more anti-cancer therapies comprise a kinase inhibitor.

103. The method of embodiment 102, wherein the kinase inhibitor is an inhibitor of EGFR, c- Met, HER2, HER3, HER4, VEGFR or AXL.

104. The method of embodiment 103, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib. 105. The method of embodiment 94, wherein the prior anti-cancer therapy is chemotherapy.

106. The method of embodiment 94, wherein the prior anti-cancer therapy is a targeted anti-cancer therapy.

107. The method of embodiment 101, wherein the one or more anti-cancer therapies comprise chemotherapy.

108. The method of embodiment 101, wherein the one or more anti-cancer therapies comprise a targeted anti-cancer therapy.

109. The method of any one of embodiments 75-108, wherein the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered essentially concurrently to at two different injection sites.

110. The method of any one of embodiments 75-108, wherein the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are combined prior to administration and administered simultaneously.

111. The method of any one of embodiments 75-108, wherein the bispecific anti-EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered sequentially over the course of one or more days as part of a multi-step treatment regimen.

112. A method of treating a solid tumor, ameliorating cancer progression, enhancing immune cell infiltration into a solid tumor, or reducing lactate production in the tumor microenvironment in a subject in need thereof, the method comprising administering a bispecific anti-EGFR/c-Met antibody to the subject, wherein the subject has received a prior administration of the PD-(L)1 axis inhibitor.

113. The method of embodiment 112, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

114. The method of embodiment 113, wherein (a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

115. The method of embodiment 113, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

116. The method of embodiment 113, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

117. The method of embodiment 112, wherein the prior-administered PD-(L) 1 axis inhibitor is an inhibitory antibody.

117a. The method of embodiment 112, wherein the PD-(L)1 axis inhibitor is an anti-PD-1 antibody.

118. The method of embodiment 117, wherein the prior-administered inhibitory antibody is selected from the group consisting of pembrolizumab (KEYTRUDA®), cetrelimab, nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

119. The method of any one of embodiments 112-118, wherein the immune cells are selected from the group consisting of T cells, B cells, and natural killer cells.

120. The method of embodiment 119, wherein the immune cells comprise CD8+ T cells and CD4+ T cells.

121. The method of embodiment 120, wherein the immune cells comprise CD8+ T cells.

122. The method of any one of embodiments 112-120, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

123. A method of reducing lactate production in the tumor microenvironment in a subject in need thereof, the method comprising administering a combination of inhibitory agents to the subject, wherein the combination of inhibitory agents is selected from the group consisting of: (a) a combination comprising an inhibitor of EGFR, an inhibitor of c-Met and a PD-(L)1 axis inhibitor; and

(b) a combination comprising an inhibitor of EGFR and an inhibitor of c-Met, wherein the subject has received a prior administration of a PD-(L)1 axis inhibitor.

124. The method of embodiment 123, wherein the combination of an inhibitor of EGFR and an inhibitor of c-Met comprises a bispecific anti-EGFR/c-Met antibody.

125. The method of embodiment 124, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

126. The method of embodiment 125, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

127. The method of embodiment 124, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

128. The method of embodiment 124, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

129. The method of embodiment 124, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

129a. The method of embodiment 124, wherein the PD-(L)1 axis inhibitor is an anti-PD-1 antibody.

130. The method of embodiment 129, wherein the inhibitory antibody is selected from the group consisting of pembrolizumab (KEYTRUDA®), cetrelimab, nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

131. The method of any one of embodiments 123-130, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

132. The method of any one of embodiments 123-131, wherein the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

133. The method of any one of embodiments 123-132, wherein the cancer is resistant to treatment with PD-(L)1 axis inhibitors.

134. The method of any one of embodiments 123-133, wherein the cancer is selected from the group consisting of head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), or non-small cell lung cancer (NSCLC).

135. A kit comprising a first pharmaceutical composition comprising a bispecific anti-EGFR/c-Met antibody and a second pharmaceutical composition comprising a PD-(L)1 axis inhibitor in two or more containers.

136. The kit of embodiment 135, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12. 137. The kit of embodiment 135, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region

(VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ IDNO: 16.

138. The kit of embodiment 135, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

139. The kit of embodiment 135, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

140. The kit of embodiment 135, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

140a. The method of embodiment 135, wherein the PD-(L)1 axis inhibitor is an anti-PD-1 antibody.

141. The kit of embodiment 140, wherein the inhibitory antibody is selected from the group consisting of pembrolizumab (KEYTRUDA®), cetrelimab, nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

142. The kit of embodiment 141, wherein the Pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26.

142a. The kit of embodiment 141, wherein the Pembrolizumab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 27, and a light chain variable region (VL) of SEQ ID NO: 28.

143. The kit of embodiment 141, wherein the Pembrolizumab antibody comprises a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO: 30.

144. The kit of embodiment 141, wherein the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36.

144a. The method of embodiment 141, wherein the cetrelimab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 37, and a light chain variable region (VL) of SEQ ID NO: 38.

145. The kit of embodiment 141, wherein the cetrelimab antibody comprises a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

146. The kit of any one of embodiments 135-145, wherein the first pharmaceutical composition comprising the bispecific anti-EGFR/c-Met antibody further comprises a first pharmaceutically acceptable excipient and wherein the second pharmaceutical composition comprising the PD-(L)1 axis inhibitor further comprises a second pharmaceutically acceptable excipient

[000188] The present invention will now be described with reference to the following specific, non-limiting examples.

Example 1. Combinatorial activity of Amivantamab and Pembrolizumab in head and neck squamous cell carcinoma and lung squamous cell carcinoma expressing wild-type EGFR and MET

[000189] Amivantamab is a bispecific human immunoglobulin (Ig)Gl Duobody® antibody that binds epidermal growth factor receptor (EGFR) and mesenchymal-epithelial transition (cMet) receptor.

[000190] To further explore its antitumor activity in EGFR wild-type head & neck cancer , the efficacy of amivantamab as a single agent and in combination with pembrolizumab was studied in head & neck cancer patient-derived xenograft (PDX) model YHIM-3003.

[000191] While amivantamab or pembrolizumab alone only moderately delay tumor growth in this model, the combination of amivantamab (10 mg/kg, BIW) and pembrolizumab (10 mg/kg, Q5D) inhibited the growth of YHIM-3003 tumors significantly at the end of the study while all regimens including the combo were well tolerated during the entire course of the treatment.

[000192] The instant study demonstrates the combinatorial benefits of amivantamab and pembrolizumab by effectively remodeling the tumor immune microenvironment, providing a strong preclinical rationale to clinically combine amivantamab and PD-1 blockade treatments. [000193] The materials and methods used for the experiments are now described.

Test Agents and Controls

[000194] Amivantamab: Amivantamab was diluted with anti-septic grade of HBSS solution for intra-peritoneal injection at 10 mg/kg per mouse.

[000195] Pembrolizumab (anti-PD-1): anti-PD-1 was diluted with anti-septic grade of HBSS solution for intra-peritoneal injection at 10 mg/kg per mouse.

[000196] Test article formulation was prepared weekly and divided into aliquots for each dispensation. Prepared aliqouts were stored at 4°C until required.

Patient derived xenograft selection.

[000197] PDX library tumors were screened for EGFR and MET expression by immunohistochemistry (Figure 1). Immunohistochemical staining of EGFR and MET were quantified using intensity scores and compared between the models prior to selecting the tumors with dual expression of EGFR and MET among LUSC and HNSCC. By intensity score, HNSCC model (YHIM-3003) showed co-expression of EGFR (average score of 157.8 + 21.3) and MET (average score of 21.4 + 12.6). LUSC model (YHIM- 2010) showed both expression of EGFR (average score of 199.4 + 48.82) and MET (average score of 161.4 + 34.1). LUSC model was previously deemed insensitive to treatment with pembrolizumab (Figure 2A and 2B). Additionally, YHIM-3003 and YHIM- 2010 demonstrated the most consistent and stable tumor development compared to the other models.

Animal Models

[000198] PDX mouse models were created using 6 to 8 weeks old female SCID (NOG) obtained from Orient Bio. After removal of the necrotic and supporting tissues from core biopsy specimens, small specimens of the tumor tissue (3 mm x 3 mm x 3 mm) from each patient were implanted subcutaneously in 1 to 2 mice. Tumor was surgically removed after the tumor exceeded 1.5 cm in diameter. The tumor was dissected into small pieces (3 mm x 3 mm x 3 mm) and reimplanted into hCD34 humanized mice Jackson Laboratories, Sacramento, CA, USA).

[000199] All animals were monitored and examined in detail for general health condition. Humanized mice were also screened for infections. The animals were housed in groups of 5 in individually ventilated cages in quarantine area for a week. Healthy animals were selected and transported into the study room.

[000200] Procedures involving the care and use of animals in the study were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC number, 2021-0112) prior to conduct. During the study, the care and use of animals were in accordance with the principles outlined in the Guide for the Care and Use of Laboratory Animals, 8th Edition, 2010 (National Research Council).

[000201] Treatment of animals was terminated, and animals humanely sacrificed, if animals were found moribund with severe clinical signs or more than 15% drop in body weight in a day or more than 20% drop in body weight from pre-test level.

Table 1: In Vivo Study Design

Group Treatment Route, Dosing schedule

Amivantamab Efficacy Study in YHIM-3003 (n = 10 mice per group)

1 Vehicle

2 Pembrolizumab (anti-PD-1, i.p., Q5D

3 ^10mS^/kS⁾ i.p., BIW

4 Amivantamab (10 mg/kg) . g^y f_or amivantamab; j p QSD for

Amivantamab + pembrolizumab pembrolizumab

Table 2: LUSC Study Design

Table 3: HNSCC Study Design

Table 4: LUSC Study Design

Sample preparation

[000202] Blood, tumor and spleen samples were collected at Day 5 of the in vivo experiments. The tumor samples were excised into portions. A proportion of tumor tissue was fixed in 10% formalin and made into FFPE blocks to generate slides for multiple IHC. The remaining proportion was divided to be used for single cell RNA sequencing and flow cytometry. Spleen samples were processed for FFPE blocks and flow cytometry. For flow cytometry analysis, the tumor was chopped into small pieces using sterile surgical blades in PBS and enzymatically dissociated into single cells with collagenase (Worthington Biochemical, New Jersey, US) at 37°C for one hour and filtered through 70 pm cell strainer. Spleen samples were dissociated by gently tapping with syringe stopper in PBS before filtering through 70 pm cell strainer. Dissociated samples were frozen in FBS + 10% DMSO stored at -80°C until required for analysis.

In vivo drug treatment and preparation

[000203] Drug treatment initiated when the tumor reached 200 mm3 for both models. Amivantamab was administered at 10 and 30 mpk (i.p., BIW) in combination with pembrolizumab (10 mpk, i.p., Q5D) for the LUSC PDX (YHIM-2010). For the duration of HNSCC PDX (YHIM-3003) experiment, 10 mpk of amivantamab was administered in combination with pembrolizumab. The treatment groups consisted of vehicle, amivantamab, pembrolizumab and combination. Each group consisted of 15 mice where 5 mice were ethically sacrificed at Day 5 and the remaining 10 mice persisted with treatment until the end of survival assay. Amivantamab and pembrolizumab were prepared by diluting with anti-septic grade of Hanks’ Balanced Sat Solution (HBSS, Life Technologies, NY, USA) solution for intra-peritoneal injection. The drug solution was prepared on the day and mixed vigorously before injection.

Multiplex immunohistochemistry

[000204] IHC was performed using the Automated Staining System (BOND Rx, Leica Biosystems). Briefly, 4-mm paraffin-embedded tumor sections were deparaffinized and rehydrated. Slides then underwent heat-induced epitope retrieval with citrate buffer at 100°C for 20 minutes. Antibodies were used at 1 : 100 dilution and hematoxylin solution were used for counter-staining. Stained slides were visualized with a Vectra Polaris and the Phenochart program. Averages of H-score were calculated for whole slides images of IHC and analysed for statistical significance of p-value (less than 0.05). The targets for T lymphocyte panel consisted of PD-1, GZMB, FOXP3, CD4, CD8 and pan-CK with detection fluorescence at 620nm, 570nm, 520nm, 690nm, 480nm and 780nm, respectively (Table 5). The targets for Myeloid panel composed of CD16, PD-L1, CD163, CDllc, CD68 and pan-CK at 620nm, 570nm, 520nm, 690nm, 480nm and 780nm, respectively.

Table 5: Panel 1 - Analysis of T lymphocytes

[000205] Complete slides were visualized using Vectra Automated Quantitative Pathology Imaging System (Vectra 3.0.5, PerkinElmer) and analyzed by inForm

Advanced Image Analysis Software (2.6.0) and Phenochart (1.1.0, PerkinElmer). Region of interest (ROI) was selected in the multi-stained scanned images to distinguish specific areas of the tumor tissue through a user-trained classification algorithm. Within the segmented regions of the tumor, characteristics of individual cells were identified using nuclear-based cell classification algorithm. Classified cell types were quantified and analyzed by converting cell count per area and cell count in total cells into quantitative values, represented by ‘intensity score’.

Flow cytometry analysis of immune cells

[000206] Tumor cells and splenocytes were collected at Day 5 of in vivo and analyzed with flow cytometry. For immune assay, dissociated single cells washed with FACS buffer (PBS containing 1% BSA, 0.01% Sodium Azide, 0.5mM EDTA) and blocked with FcR Blocking Reagent (Miltenyi Biotec) at room temperature for 20 minutes. Fixation and permeabilization for intracellular staining were prepared with True- Nuclear™ Transcription Factor buffer at room temperature for 30 minutes. Multi-color flow cytometric analysis was performed using BD LSRfortessa™ X-20 (BD Bioscience, New Jersey, USA). FlowJo (Flow Jo, LLC) software was used for data acquiring and analysis. Screening of tumor reactive T cells were performed using CEA and MAGEA antibody.

Cell lines

[000207] The LUSC cell lines, EBC-1 (HTB-60) and H1703 (CRL-5889), were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). The cells were cultured and maintained in HyClone™ RPMI-1640 (Cytiva, Massachusetts, US) supplemented with 10% fecal bovine serum (Cytiva) and 1% antibiotic/antimycotic solution (Cytiva) in a humidified incubator with 5% CO2.

Western blot reagents

[000208] Following primary antibodies were used for protein detection. Anti-actin (#A3854, 1 : 1000) was purchased from Sigma Aldrich. Anti-EGFR (#2232, 1 : 1000), anti- p-EGFR (#2234, 1:1000), anti-LDHA (#3582, 1:1000), anti-MET (#8198, 1:1000) and anti-p-MET (#3077, 1:1000) were purchased from Cell Signaling Technology. Anti- SLC16A3/MCT4 (ab74109, 1:1000) was purchased from Abeam. Secondary HRP- conjugated anti-rabbit (#7074, 1 : 1000) or anti-mouse IgG (#7076, 1 : 1000) purchased from Cell Signaling Technology and ECL System (#1707062, Bio-Rad) were used for protein detection.

Single-cell RNA sequencing and processing

[000209] Average body weight loss and percent changes were calculated for each group. Toxicity, if any, to treatment was valuated based on loss of body weight. All statistical calculations were performed using Prism 9.0 (Graph Pad Software Inc, USA). Statistical comparison of tumor volume and survival curves between the treatment groups was calculated using two-way ANOVA, followed by Tukey ’s multiple comparison tests (p values less than 0.05 were considered significant).

[000210] The tumor samples were dissociated using gentleMACS human tumor dissociation kit (Miltenyi Biotec) then processed prior to single cell library preparation following the manufacturer’s guidelines (10X Genomics, California, US). Volumes for each sample were calculated for target capture of 10,000 cells. The processed samples were prepared for gene expression analysis using Chromium Single Cell 5' Reagent Kits (10X Genomics). Single cell library was then outsourced for single cell RNA sequencing at Macrogen (Korea). Sequencing was performed to achieve read depth of more than 50,000. FASTQ files were then processed for mmlO and hgl9-based mapping to distinguish mouse- and human-derived genome. Read counts and merging of the samples were executed via Cell Ranger (7.1.0). Scanpy was used for further quality control including filtering (minimum of 200 genes in at least 3 cells, with <20% of mitochondrial reads as cutoff) and normalization of cells (Harmony), batch correction and clustering. Analysis of gene expression in HNSCC and LUSC models was visualized using heat maps, violin and dot plots via Seurat (version 4.9.9).

Filtering and normalization of scRNA-seq data

[000211] Four quality measures of mitochondrial genes < 20%, min genes < 200, min cells < 3 were applied to the raw gene-cell-barcode matrix of each cell. After basic filtering, patient data with less than 30 cells for each patient was excluded from the analysis. The data was scaled, and normalization was performed. The batch correction algorithm used harmony and the relative expression levels for the rest of the cells. Gene subsets were used for subsequent analysis.

Cell type identification and analysis

[000212] Primary cell type annotation using Azimuth lung database (https://azimuth_hubmapconsortium_org/) verified using cell-type signatures by CITE- seq. Each detailed cluster was divided using Seurat FindClusters algorithm. Expression of cell-type markers across clusters subsets were analysed via tSNE and UMAP. Top 50 genes of hetergenous EGFR and MET high tumor clusters were analysed for top 50 differental gene expression (DEG) markers. Among analysed genes, key factors of immunomodulatory were marked in volcano plot. Tumor associated tetramer assay

[000213] Tetramer peptides were generated to compare the proportion of tumor specific T lymphocytes between the treatment groups. MAGEA2 (HLA-A*24:02 EYLQLVFGI) and CEA (HLA-A*24:02 TYACFVSNL) were outsourced from Biolegend (California, US). Preparation of Flex-TTM Tetramer is available on the Internet for example at the Biolegend web site.

Calculations

[000214] Tumor volume was calculated using the formula: Tumor Volume (TV, mm3) = (L*W2/2); where ‘L’ is the tumor length (longest tumor dimension), and ‘W’ is the tumor width (the longest tumor dimension perpendicular to L) by caliper measurements.

[000215] The percent ATGI was defined as the difference between mean tumor burden of the treatment and control groups, calculated as:

([(TVc-TVc0)-(TVt-TVt0)]/(TVc-TVc0))xl00, where ‘TVc’ is the mean tumor burden of a given control group on a specific day, ‘TVcO’ is the mean initial tumor burden (Day 0) of a given control group, ‘TVt’ is the mean tumor burden of the treatment group on a specific day, and ‘TVtO’ is the mean initial tumor burden (Day 0) of the treatment group. [000216] In case of tumor regression, an additional metric Tumor Regression (TR) was calculated as: (1-mean (TVt / TV0)) x too, where TVt and TV0 are defined the same as above.

Data Analysis

[000217] In each study, statistical comparisons were made in reference to the vehicle group, up to and including the last day of the study when all 10 mice remained in each group. Differences between groups were considered significant when p<0.05.

Statistical significance for tumor volume and for body weight was calculated using the linear mixed-effects analysis in R software Version 4.0.3 (using Janssen’s internally developed Shiny application In Vivo Longitudinal Data Analysis Version 4.8), with treatment and time as fixed effects and animal as random effect with time treated as a continuous variable (Pinheiro J, Bates D. Mixed-effects models in S and S-Plus. Heidelberg, Germany: Springer; 2000). Logarithmic transformation (Base 10) was performed if individual longitudinal response trajectories were not linear. The information derived from this model was used to make pairwise treatment comparisons to the control group or between all the treatment groups over time. [000218] The experimental results are now described.

Antitumor efficacy of amivantamab in combination with pembrolizumab in head and neck squamous and lung squamous humanized mice models.

[000219] EGFR and MET-expressing tumors obtained from a head and neck squamous cell carcinoma (HNSCC) and a lung squamous cell carcinoma (LUSC) patient were transplanted into Hu-CD34-NSG to establish humanized patient-derived xenograft (PDX) models. Tumor models characteristics are shown in Table 6.

Table 6. PDX models

[000220] Tumor-bearing PDXs were treated with vehicle, pembrolizumab (lOmpk, Q5D, n=10), amivantamab (lOmpk, BIW, n=10), or a combination of pembrolizumab and amivantamab (n=10) (Figure 2A). Analysis of immune modulatory responses within the tumor microenvironment (TME) using multiplexed immunohistochemistry, flow cytometry, and single cell RNA sequencing was performed.

[000221] Combination of amivantamab and pembrolizumab showed a significant reduction of tumor volume (p<0.001) compared to vehicle or single arm treatment in both HNSCC and LUSC models (Figures 3-5). Synergistic benefits of amivantamab and pembrolizumab combination treatment compared to monotherapy were observed in the in vivo experiment of HNSCC (YHIM-3003) and LUSC (YHIM-2010) tumor bearing PDX models. In the HNSCC PDX, combination of amivantamab and pembrolizumab treatment showed a significant reduction of tumor volume compared to vehicle or single arm treatments (p<0.001, Figure 3A-3B). The greatest tumor growth inhibition (TGI) rate was observed largely in the combination treated group (Figure 3B). TGI over 40% was observed in 70% of combination group whereas only 20% of amivantamab group was greater than 40% TGI (Figure 3B). TGI did not exceed over 40% in the pembrolizumab group. [000222] Additionally, significantly longer survival was observed for combination treated compared to the vehicle treated groups (pO.OOOl). Additionally, longer survival was observed for the combination treatment group compared to the monotherapy groups. At the end of Day 19, 40% of combination group had survived in contrast to pembrolizumab and amivantamab monotherapy where all the mice demised at Day 17 and 19, respectively (Figure 3A).

[000223] In the LUSC PDX, amivantamab administered at the same dosage of 30 mpk in combination with pembrolizumab significantly reduced tumor volume compared to pembrolizumab group (p<0.001, Figure 5). Interestingly, no rebound of tumor growth was observed in amivantamab and combination group after terminating drug administration at Day 47 (Figure 5). It appeared that amivantamab boosted immune activity of lymphocytes including memory phenotypic subsets of T cells and prevented recurrence of tumor development after the treatment has been terminated. However, amivantamab at 30 mpk alone effectively reduced tumor volume (Figure 5). This indicated that the dose of amivantamab needed to be decreased in order to evaluate the combination synergy with pembrolizumab. Amivantamab given at a modified dosage of 10 mpk in combination with pembrolizumab significantly reduced the tumor volume compared to amivantamab and pembrolizumab monotherapy groups (p<0.05 and p<0.001, respectively, Figure 4). Combination treatment also resulted in improved survival over 61 days. All mice in the vehicle and pembrolizumab group demised at Day 47 and 53, respectively, where 70% and 40% survived until Day 61 in the combination and amivantamab group, respectively (Figure 4).

Tumor infiltrating CD8+ T cells were significantly higher in the combination-treated tumors.

[000224] Multispectral imaging of tumor indicated that granzyme B-producing CD 8+ T cells were significantly increased within the tumor in the combination group (p<0.01) (Figures 6-7). Landscape of the tumor microenvironment (TME) within the HNSCC PDX tumor revealed that the infiltration of immune cells in the tumor nest and stromal sites was enhanced by combination treatment of amivantamab and pembrolizumab (Figure 6). Proportion of granzyme B expressing (GZMB) CD8 T cells in tumor nest of TME was significantly increased in the combination group compared to the control group (18.81 ± 10.36 and 4.09 ± 1.55, respectively, p<0.05, Figure 7). There were no statistical differences between the groups in the stroma, however, an increasing trend was observed in the combination group. In general, GZMB+ CD8 T cell population increased (in the total area of TME) by combination treatment of amivantamab and pembrolizumab in YHIM-3003. In contrast, no profound results or trends were observed in the proportion of Foxp3 expressing regulatory CD4 T cells between the treatment groups. These results suggested that amivantamab and pembrolizumab synergistically enhanced infiltration of active cytotoxic CD8 T cells into the TME and suppressed growth of tumor cells.

Anti-tumor phenotypic changes in T cell populations induced by combination treatment

[000225] Further analysis of T cell subsets suggested that central memory type CD 8+ T cells were increased upon combination treatment. This group also demonstrated significantly higher CEA -tetramer positive CD8+ T cells in the tumor (p < 0.01), suggesting that cytotoxic T cells recognizing tumor specific antigens enhanced antitumor immune response (Figures 8-9).

[000226] To compare the changes in the populations of lymphocytes between the treatment group, phenotypic alterations of immune cells were analyzed using flow cytometry. Combination treatment group had significantly increased proportion of central memory CD8 T cells compared to the single treatment of pembrolizumab in HNSCC PDX (13.38 ± 8.80 and 2.11 ± 2.34 respectively, p<0.05, Figure 8 A) and LUSC PDX (24.00 ± 8.26 and 10.55 ± 4.47 respectively, p<0.05, Figure 8B). Analysis of T cells identified a shared trend between the HNSCC and LUSC PDX models in the central memory subset of cytotoxic CD8 T cells (Figure 8C). No recurrence of tumor development was observed after the treatment of amivantamab at high dosage in combination with pembrolizumab (Figure 5). This observation initially suggested that combination treatment may stimulate memory subsets of T cells during immune response to tumor development. Immune profiling of humanized PDX tumor indicated that combination of amivantamab and pembrolizumab enhanced induction of memory subpopulations of T cells (i.e., central memory CD8 T cells, Figures 3A-3C.

[000227] Splenocytes of YHIM-3003 mice were processed and stained with tetramer antibodies to detect T lymphocytes that can recognize the tumor specific antigens CEA (carcinoembryonic antigen). The combination group had significantly higher population of CEA-expressing tumor reactive CD8 T cells compared to the vehicle group (8.28 ± 2.67 and 3.02 ± 0.75, respectively, p<0.05, Figure 9). The combination group also had the highest proportion compared to amivantamab (6.17 ± 1.98) and pembrolizumab (6.20 ± 3.05, Figure 9). Pembrolizumab monotherapy induces an EGFR/MET high subcluster that upregulates genes implicated in lactate production and immune suppression.

[000228] Single cell RNA sequencing analysis of HNSCC tumors showed that an EGFR^hlghMET^high cluster was enriched in the TME after pembrolizumab treatment. EGFR^hlghMET^high subcluster showed elevated glycolysis and lactic acid pathway -related genes compared to EGFR^lowMET^low cluster. Lactate transporter, MCT4 (SLC16A3) and LDHA genes were dramatically increased in the EGFR^I"-'^I'MET¹"-'¹' cluster (Figures 10-19). Elevated lactic acid pathway may lead to immune evasion in the tumor, dampening the activity of pembrolizumab. Interestingly, combination treatment with amivantamab could reduce EGFR^I"-'^I'MET¹"-'¹' subcluster, and could effectively control tumor via creating favorable immune TME.

[000229] Specifically, in HNSCC PDX tumor, a subcluster with high expression of EGFR simultaneously had elevated level of MET (Figure 12). Interestingly, single treatment of pembrolizumab induced higher density of EGFR and MET dual expressing subcluster compared to the other treatment groups (Figure 12). The tumor subcluster expressing high levels of both EGFR and MET (EMHIGH) was define and analyzed for top 50 genes to identify differentially expressed genes (DEGs) in EGFR^hlgh/MET^hlghand EGFR^LOW/MET^LOW tumor subclusters (Figure 13 and 14). Among the top genes, ANXA1, ARF1, HLA-E, LDHA, SLC16A3, S100A11 and TPT1 were key immunomodulatory factors that showed significant fold changes, with LDHA and SLC16A3 showing 128 and 25 log fold changes, respectively (p<0.05 with log2 fold change >1, Figure 14), indicating that these markers were significantly elevated in the EGFR^l"^gl7MET^l'^lgl' subcluster compared to the EGFR^LOW/MET^LOW subcluster. These markers were also key factors in the EGFR^HIGH subcluster of the LUSC PDX tumor (data not shown). In this study, lactate dehydrogenase A (LDHA) and SLC16A3 were revealed as core genes with regulatory functions in the biological process of glucose metabolism that may hinder immune surveillance and responses within the TME. The elevation of EGFR in the HNSCC PDX treated with pembrolizumab was also confirmed by multiplex IHC (Figure 18). EGFR intensity in the tumor of pembrolizumab group was found to have an intensity score of 699 ± 31.6, which was significantly higher than the score of vehicle, amivantamab and combination group (180 ± 33.1, 88 ± 9.5, 182 ± 29.4, respectively, p<0.01, Figure 19). This suggested that a large proportion of the tumor actively upregulated the expression of EGFR and MET in one of multifaceted responses to pembrolizumab. It is feasible that upregulation of EGFR and MET in tumor cells contributed to resistance to pembrolizumab monotherapy (Figure 3A). Amivantamab downregulates genes induced by EGFR/MET high subcluster and promotes antitumor immunity

[000230] Based on the public database of The Cancer Genome Atlas (TCGA), expression of EGFR in both HNSCC and LU SC significantly correlated with the expression of LDHA and SLC16A3 (p<0.05, Figure 17A), suggesting that regulation of EGFR and glucose metabolism simultaneously contribute to TME favoring progression of tumor. In this study, expression of LDHA and SLC16A3 was significantly higher in the EGFRHIGH/METHIGH subcluster of HNSCC PDX tumor compared to the EGFRLOW/METLOW subcluster (p<0.05, Figure 17B). Additionally, the expression of LDHA and SLC16A3 was notably higher in the pembrolizumab group. (Figure 17B). Transcriptional upregulation of other glycolytic markers was also predominantly increased in the EGFRHIGH/METHIGH subcluster and the pembrolizumab group (HK2, GPI, ALDO A, PGK1, PGAM1, ENO1 and ENO2, Figure 17C). Moreover, key regulators of hypoxia showed the same trend in the EGFRHIGH/METHIGH subcluster and the treatment groups (HIF1A, HDAC1, KDM1A, KDM2A, CA9, VEGFA and TWIST1, Figure 17D), suggesting that the tumor in the HNSCC PDX mice treated with pembrolizumab feasibly had an acidic environment by favoring anerobic metabolism of glucose in the TME.

[000231] Correlation of translational and surface expression of EGFR and MET was also confirmed in HNSCC and LUSC cell lines. T cells secrete IFN-y that functions as autocrine/paracrine molecule in response to pembrolizumab in the tumor site. Thus, cell lines were treated with IFN-y to replicate the immunological stimulus in the TME of PDX models by treatment of pembrolizumab. At the protein level, upregulation of EGFR and MET in selected HNSCC and LUSC cell lines after IFN-y showed a moderate correlation, while expression of pEGFR and pMET manifested a strong positive correlation (r = 0.9824, pO.OOOl, Figure 17E). In addition, IFN-y-induced surface expression of EGFR and MET showed positive correlation (r = 0.9507, p<0.005, Figure 17E). In H1703, upregulation of EGFR and MET, and pEGFR and pMET by IFN-y was inhibited by amivantamab. Amivantamab also reduced the expression of SLC16A3 (monocarboxylate transporter 4, MCT4) in H1703 in the presence of IFN-y (Figure 17F). Amivantamab also reduced immune checkpoint related markers including PD-L1 in the EGFR^HIGHMET^HIGH tumor subcluster.

[000232] DEG analysis of HNSCC PDX tumor illustrated that MET-regulated genes and other immune checkpoint markers were also increased in the EGFR^HIGH/MET^HIGH tumor subcluster. Significantly increased MET-related genes were BACE2, CD274, CD276, DPYD, PRSS23, PYGL, STK40 and S100A4 (Figure 20A). In addition to elevated MET-regulated genes, MET-STAT4-PD-L1 axis had noticeably increased expression in the EGFR^HIGH/MET^HIGH tumor group (Figure 20B). It appeared that expression of these markers had decreased in the combination group, suggesting that amivantamab and pembrolizumab combination impeded downstream signaling of MET- regulated genes (Figure 20C).

Effect of treatment with Amivantamab alone, or in combination with Pembrolizumab on efficacy and Body Weight in YHIM-3003 PDX Model [000233] In PDX model YHIM-3003, the efficacy of Amivantamab (10 mg/kg, i.p., BIW), pembrolizumab (10 mg/kg, i.p., Q5D), or combination was evaluated for 19 days. No bodyweight loss (Figure 21 A) or adverse clinical signs were observed during the dosing period, indicating that amivantamab and pembrolizumab treatments were well tolerated at the indicated dosing regimen.

[000234] Amivantamab or pembrolizumab mono-treatment had minor impact on the tumor growth rate, with the percentage ATGI of 15.64% and 10.26% on day 19, respectively (Figure 2 IB, Table 7) in PDX model YHIM-3003. In contrast, the combination of Amivantamab and pembrolizumab induced stronger tumor growth inhibition with ATGI of 57.37% (p value <0.0001) compared with the vehicle control.

Table 7: Tumor Growth Inhibition and p Values of YHIM-3003 PDX model on day 19.

Treatment ATGI p value Significance

Vehicle NA NA NA

Amivantamab (10 mg/kg) 15.64 0.0670

Pembrolizumab (10 mg/kg) 10.26 0.1440

Amivantamab + pembrolizumab 57.37 <0.0001 ***

NA, not applicable; TGI, tumor growth inhibition; TR, tumor regression.

Percent ATGI was calculated on day 28 compared. Significance values were calculated over time to day 28. [000235] The goal of this study was to evaluate the antitumor activity of amivantamab as a monotherapy or delivered in combination with anti-PD-1 antibody pembrolizumab in a humanized patient-derived xenograft model. Although amivantamab or pembrolizumab alone only moderately delay tumor growth in this model, the combination of amivantamab (10 mg/kg, BIW) and pembrolizumab (10 mg/kg, Q5D) inhibited the growth of YHIM-3003 tumors significantly at the end of the study while all regimens including the combo were well tolerated during the entire course of the treatment.

Discussion

[000236] This study demonstrated combinatorial benefits of amivantamab and pembrolizumab by effectively remodeling the tumor immune microenvironment, providing a strong preclinical rationale to clinically combine amivantamab and PD-1 blockade treatments. In fact, there is an ongoing study of combination therapy with amivantamab and cetrelimab in patients with metastatic NSCLC (PolyDamas, NCT05908734), and the clinical results are awaited.

[000237] The combination synergy of amivantamab and pembrolizumab was observed in the treatment of humanized PDX mouse model bearing, pembrolizumab- insensitive HSNCC (YHIM-3003) and LUSC (YHIM-2010) tumors. By comprehensive analysis of immunogenic mechanism behind the combination synergy, we noted that the combination treatment enhanced stimulation of central memory subset of cytotoxic CD8+ T cells in both PDX models, and induced tumor-reactive T cells in HNSCC PDX. Central memory subset of CD8+ T cells have shown superior in vivo and in vitro antitumor immunity compared with effector memory T cells. In addition to modulating T cells, combination treatment increased proportion of granzyme B-expressing cytotoxic CD8+ T cells in the TME, suggesting that combination treatment encouraged infiltration of physiologically active CD8+ T cells into the tumor nest. These data suggest that combination treatment can ultimately enhance T-cell mediated tumor killing in the tumors that were previously non-responsive to immunotherapy.

[000238] We subsequently analyzed the single cell transcriptomic landscape of the tumor and revealed that tumor cells exhibited relatively high EGFR and MET expression in response to pembrolizumab treatment alone. EGFR signaling in cancers has been associated with global metabolism favoring highly glycolytic tumors. We demonstrated that EGFR^HIGH tumor subcluster was distinctly elevated in the pembrolizumab-treated group and exhibited significantly higher level of lactate producing biomarkers including LDHA and SLC16A3 compared to the combination group in both HNSCC and LUSC PDX.

[000239] The findings of this study suggest that pembrolizumab may promote a tumor-immune microenvironment with metabolic characteristics of the ‘Warburg phenotype’ in the tumor. LDHA is essential for conversion of pyruvate into lactate while SLC16A3 facilitates exchange of lactate between the cells and extracellular matrix (ECM). It is likely that the EGFR^HIGH tumor cells produced and actively exported lactate into the ECM of TME. Lactate provides metabolic fuel for cancer cells as well as tumor-killing immune cells such as T and NK cells, however, accumulation of lactate due to enhanced glycolysis greatly disables the ability of CD8+ T and NK cells to infiltrate into the tumor site. Several studies have also highlighted that lactate suppressed cytotoxic activity of CD8+ T cells by inhibiting production of IFN-y, which is crucial for facilitating tumor killing by CD8+ T and NK cells. This suggests that the tumor with high EGFR expression may be unresponsive to immunotherapy by disturbing essential mechanism and subsequent loss of functionality in cytotoxic T and NK cells in the TME. In fact, the single-agent activity of pembrolizumab alone in HNSCC, which expresses high levels of EGFR and MET, has an objective response rate of 16.9%, with the median progression- free survival of 2.3 months.

[000240] In addition to creating metabolic environment that favors tumor persistence, we found that pembrolizumab treatment also increased MET-related immune checkpoint markers and expression of PD-L1. It appeared that pembrolizumab treatment alone induced upregulation of immune checkpoint in the tumor cells, contributing to becoming more resistant to T cell killing and evasion of the immune response via enhanced signaling through MET-STAT4-PD-L1 axis. We hypothesize that the HNSCC PDX model became insensitive to anti-PD-1 immunotherapy alone by developing multifaceted bypassing mechanisms, upregulating MET signaling pathway in addition to amplification of EGFR. These data suggests that amivantamab diminishes the immunosuppressive effects by pembrolizumab on the immune cells in the TME. Significant reduction of glycolytic and MET-regulated immune checkpoint biomarkers in the combination treatment group restored infiltration and activation of infiltrating cytotoxic CD8 T and NK cells, which was also evident in the in vivo models and comprehensive immune profiling analysis. These phenomena were less evidently seen in the lung squamous cell carcinoma model, as there was less abundant co-expression of EGFR and MET in the tumor. However, gene expression analysis showed similar trend in the EGFR and MET. [000241] Upon binding with major histocompatibility complex molecules, T cells secrete IFN-y to enhance anti-tumor activity of surrounding T lymphocytes. IFN-y signalling also promotes MET activation and induce immune checkpoints via enhanced MET-STAT4-PD-L1 axis in tumor cells, providing tumor cells with immune evasion mechanism. This was also evident in our study that MET-STAT4-PD-L1 axis and MET- related immune checkpoints were elevated particularly in the pembrolizumab-treated EGFR^HIGH tumor. These alterations are likely to facilitate tumor growth by allowing immune tolerance and may affect the response to immune checkpoint inhibitors.

[000242] In conclusion, this study revealed that amivantamab diminished the immunosuppressive effects induced by pembrolizumab in the EGFR^HIGHMET^HIGH tumor of pembrolizumab insensitive humanized HNSCC PDX model. Combination of amivantamab and pembrolizumab significantly reduced tumor volume in HNSCC and LUSC tumor bearing PDX model compared to amivantamab or pembrolizumab alone. This study demonstrated that combination treatment greatly enhanced antitumor phenotypic changes in infiltrating CD8 T cells and central memory T cells in the TME. Single cell RNA transcriptomic analysis showed that pembrolizumab alone induced tumor invasive mechanism by upregulation of EGFR and MET signaling. However, the same biomarkers were reduced when pembrolizumab was administered in combination with amivantamab. This study highlighted rationale for combination therapy of amivantamab and PD-1 blockade immunotherapy that can be applied in clinical treatment regimen of solid advanced tumors.

[000243] Overall, the data suggest that amivantamab in conjunction with a PD-(L) 1 axis inhibitor, such as pembrolizumab, may offer synergistic anti-tumor efficacy in the EGFR and MET wild-type head & neck cancers in which anti-PD-1 treatment has limited benefit. Our study demonstrated combinatorial benefits of amivantamab and a PD-(L)1 axis inhibitor, such as pembrolizumab, by effectively remodeling TME, providing a rationale to clinically combine amivantamab and PD-(L)1 axis inhibitors.

Example 2. Amivantamab, an EGFR-MET bispecific antibody, in combination with cetrelimab, an anti-PD-1, in advanced non-small cell lung cancer: The phase 1/2 PolyDamas study

[000244] Background: Patients with advanced non-small cell lung cancer (NSCLC) harboring EGFR mutations are treated with 3rd-generation TKIs. However, EGFR activation and TKI treatment may contribute to upregulation of PD-1/PD-L1, promoting treatment resistance. Amivantamab (ami), an EGFR-MET bispecific antibody with immune cell-directing activity, is effective against EGFR-mutated advanced NSCLC as monotherapy or in a combination. Cetrelimab (cet) is an anti-PD-1 monoclonal antibody with clinical activity in previously treated NSCLC (Felip Cancer Chemother Pharmacol 2022;89(4):499-514). Simultaneous targeting of the innate and adaptive immune systems with ami and cet may improve antitumor activity versus either agent alone. The PolyDamas study (NCT05908734) aims to identify the recommended phase 2 combination dose (RP2CD) and evaluate the antitumor effect of ami+cet in patients with advanced NSCLC.

[000245] Methods: This open-label, multicenter, interventional study will have a combination dose selection phase, followed by an expansion phase. The primary endpoints include safety (phase 1 dose selection) and objective response rate by investigator using RECIST 1.1 criteria (phase 2 dose expansion). For dose selection, 20 patients with advanced NSCLC, with or without known driver mutations, who have progressed on or after standard of care anti-cancer therapy, will be enrolled. Dosing will be in 28-day cycles, starting with a reduced ami dose (700 mg IV; 1050 mg if >80 kg) given once weekly (initial dose as a split infusion over the first 2 days) for the first 4 weeks (cycle 1), then every 2 weeks (Q2W) thereafter (cycle 2 onwards). Cet will be dosed IV 240 mg Q2W, with the first dose given on day 2 of cycle 1. Dose escalation/de-escalation will be based on the observation of dose-limiting toxicities. The RP2CD will be selected through a Bayesian optimal interval design with a 3+3 design run in.

[000246] The phase 2 expansion cohorts will enroll 30 patients each after identification of the RP2CD. Cohort A will emoll patients with advanced NSCLC harboring EGFR exon 19 deletion or L858R mutation and have had disease progression on a 3rd-generation TKI and platinum-based chemotherapy. Cohort B will emoll patients with treatment-naive, wild-type advanced NSCLC (no known driver mutations) and PD- L1 tumor score >50%. The phase 2 secondary endpoints include duration of response, disease control rate, progression-free survival, and overall survival.

[000247] This study is exploratory in nature and hypothesis-generating; no formal statistical hypotheses exist for either phase. In both phases, patients will continue study treatment until disease progression, unacceptable toxicity, or until another criterion for treatment discontinuation is met. This study is currently enrolling, with an enrollment goal of 80 patients total.

Claims

We Claim:

1. A method of treating a solid tumor or ameliorating cancer progression in a subject in need thereof, the method comprising administering to the subject:

(a) a PD-(L)1 axis inhibitor; and

(b) a bispecific anti-EGFR/c-Met antibody.

2. The method of claim 1, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

3. The method of claim 2, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

4. The method of claim 2, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

5. The method of claim 4, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

6. The method of claim 1, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

7. The method of claim 6, wherein the inhibitory antibody is selected from the group consisting of cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

8. The method of claim 7, wherein the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36.

9. The method of claim 7, wherein the the cetrelimab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 37, and a light chain variable region (VL) of SEQ ID NO: 38; or a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

10. The method of claim 7, wherein the pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26.

11. The method of claim 7, wherein the pembrolizumab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 27, and a light chain variable region (VL) of SEQ ID NO: 28; or a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO: 30.

12. The method of claim 1, wherein the cancer is an EGFR, c-Met or EGFR and c-Met expressing cancer associated with a wild-type EGFR, an EGFR activating mutation, an EGFR gene amplification, increased levels of circulating HGF, a wild-type c- Met, a c-Met activating mutation, a c-Met gene amplification or a mutant KRAS, or any combination thereof.

13. The method of claim 12, wherein the EGFR activating mutation comprises L718Q, G719A, G719X (X being any amino acid), L861X (X being any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitution, deletion of E746-A750, deletion of R748-P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Vai and Ala (SVA) between S768 and V769, insertion of Asn and Ser (NS) between P772 and H773, insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof.

14. The method of claim 13, wherein the mutant KRAS comprises a G12V, G12C or G12A substitution.

15. The method of any one of claims 1-14, wherein the method inceases immune cell infiltration into a solid tumor, further wherein the immune cells are selected from the group consisting of T cells, B cells, and natural killer cells.

16. The method of claim 15, wherein the T cells comprise CD8+ T cells and CD4+ T cells.

17. The method of claim 16, wherein the T cells comprise CD8+ T cells.

18. The method of any one of claims 1-17, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

19. The method of any one of claims 1-18, wherein the subject is suspected to have or has an EGFR, c-Met or EGFR and c-Met expressing cancer.

20. The method of claim 1, wherein the subject is resistant or has acquired resistance to treatment with a prior anti-cancer therapy.

21. The method of claim 20, wherein the prior anti-cancer therapy is a kinase inhibitor.

22. The method of claim 21, wherein the kinase inhibitor is an inhibitor of EGFR, c-Met, HER2, HER3, HER4, VEGFR or AXL.

23. The method of claim 22, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

24. The method of any one of claims 1-23, wherein the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

25. The method of any one of claims 1-24, wherein the cancer is resistant to treatment with PD-(L)1 axis inhibitors.

26. The method of any one of claims 1-25, wherein the cancer is selected from the group consisting of head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), or non-small cell lung cancer (NSCLC).

27. The method of any one of claims 1-26, further comprising administering one or more anti-cancer therapies to the subject.

28. The method of claim 27, wherein the one or more anti-cancer therapies comprise a kinase inhibitor.

29. The method of claim 28, wherein the kinase inhibitor is an inhibitor of EGFR, c- Met, HER2, HER3, HER4, VEGFR or AXL.

30. The method of claim 29, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

31. The method of claim 20, wherein the prior anti-cancer therapy is chemotherapy.

32. The method of claim 20, wherein the prior anti-cancer therapy is a targeted anti-cancer therapy.

33. The method of claim 27, wherein the one or more anti-cancer therapies comprise chemotherapy.

34. The method of claim 27, wherein the one or more anti-cancer therapies comprise a targeted anti-cancer therapy.

35. The method of any one of claims 1-34, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered essentially concurrently to at two different injection sites.

36. The method of any one of claims 1-34, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are combined prior to administration and administered simultaneously.

37. The method of any one of claims 1-34, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L) 1 axis inhibitor are administered sequentially over the course of one or more days as part of a multi-step treatment regimen.

38. A method of enhancing immune cell infiltration into a solid tumor in a subject in need thereof, the method comprising administering to the subject:

(a) a PD-(L)1 axis inhibitor; and

(b) a bispecific anti-EGFR/c-Met antibody.

39. The method of claim 38, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

40. The method of claim 39, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

41. The method of claim 39, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

42. The method of claim 41, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

43. The method of claim 38, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

44. The method of claim 43, wherein the inhibitory antibody is selected from the group consisting of cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

45. The method of claim 44, wherein the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36.

46. The method of claim 44, wherein the cetrelimab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 37, and a light chain variable region (VL) of SEQ ID NO: 38; or a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

47. The method of claim 44, wherein the pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26.

48. The method of claim 44, wherein the pembrolizumab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 27, and a light chain variable region (VL) of SEQ ID NO: 28; or a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO: 30.

49. The method of claim 38, wherein the cancer is an EGFR, c-Met or EGFR and c-Met expressing cancer associated with a wild-type EGFR, an EGFR activating mutation, an EGFR gene amplification, increased levels of circulating HGF, a wild-type c- Met, a c-Met activating mutation, a c-Met gene amplification or a mutant KRAS, or any combination thereof.

50. The method of claim 49, wherein the EGFR activating mutation comprises L718Q, G719A, G719X (X being any amino acid), L861X (X being any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitution, deletion of E746-A750, deletion of R748-P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Vai and Ala (SVA) between S768 and V769, insertion of Asn and Ser (NS) between P772 and H773, insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof.

51. The method of claim 50, wherein the mutant KRAS comprises a G12V, G12C or G12A substitution.

52. The method of any one of claims 38-51, wherein the immune cells are selected from the group consisting of T cells, B cells, and natural killer cells.

53. The method of claim 52, wherein the T cells comprise CD8+ T cells and CD4+ T cells.

54. The method of claim 53, wherein the T cells comprise CD8+ T cells.

55. The method of any one of claims 38-54, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

56. The method of any one of claims 38-55, wherein the subject is suspected to have or has an EGFR, c-Met or EGFR and c-Met expressing cancer.

57. The method of claim 38, wherein the subject is resistant or has acquired resistance to treatment with a prior anti-cancer therapy.

58. The method of claim 57, wherein the prior anti-cancer therapy is a kinase inhibitor.

59. The method of claim 58, wherein the kinase inhibitor is an inhibitor of EGFR, c-Met, HER2, HER3, HER4, VEGFR or AXL.

60. The method of claim 59, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

61. The method of any one of claims 38-60, wherein the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

62. The method of any one of claims 38-61, wherein the cancer is resistant to treatment with PD-(L)1 axis inhibitors.

63. The method of any one of claims 38-62, wherein the cancer is selected from the group consisting of head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), or non-small cell lung cancer (NSCLC).

64. The method of any one of claims 38-63, further comprising administering one or more anti-cancer therapies to the subject.

65. The method of claim 64, wherein the one or more anti-cancer therapies comprise a kinase inhibitor.

66. The method of claim 65, wherein the kinase inhibitor is an inhibitor of EGFR, c- Met, HER2, HER3, HER4, VEGFR or AXL.

67. The method of claim 66, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

68. The method of claim 57, wherein the prior anti-cancer therapy is chemotherapy.

69. The method of claim 57, wherein the prior anti-cancer therapy is a targeted anti-cancer therapy.

70. The method of claim 64, wherein the one or more anti-cancer therapies comprise chemotherapy.

71. The method of claim 64, wherein the one or more anti-cancer therapies comprise a targeted anti-cancer therapy.

72. The method of any one of claims 38-71, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered essentially concurrently to at two different injection sites.

73. The method of any one of claims 38-71, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are combined prior to administration and administered simultaneously.

74. The method of any one of claims 38-71, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L) 1 axis inhibitor are administered sequentially over the course of one or more days as part of a multi-step treatment regimen.

75. A method of reducing glycolysis or lactic acid production in the tumor microenvironment in a subject in need thereof, the method comprising administering to the subject:

(a) a PD-(L)1 axis inhibitor; and

(b) a bispecific anti-EGFR/c-Met antibody.

76. The method of claim 75, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

77. The method of claim 76, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and (b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

78. The method of claim 76, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

79. The method of claim 78, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

80. The method of claim 75, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

81. The method of claim 80, wherein the inhibitory antibody is selected from the group consisting of cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

82. The method of claim 81, wherein the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36.

83. The method of claim 81, wherein the cetrelimab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 37, and a light chain variable region (VL) of SEQ ID NO: 38; or a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

84. The method of claim 81, wherein the pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26.

85. The method of claim 81, wherein the pembrolizumab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 27, and a light chain variable region (VL) of SEQ ID NO: 28; or a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO: 30.

86. The method of claim 75, wherein the cancer is an EGFR, c-Met or EGFR and c-Met expressing cancer associated with a wild-type EGFR, an EGFR activating mutation, an EGFR gene amplification, increased levels of circulating HGF, a wild-type c- Met, a c-Met activating mutation, a c-Met gene amplification or a mutant KRAS, or any combination thereof.

87. The method of claim 86, wherein the EGFR activating mutation comprises L718Q, G719A, G719X (X being any amino acid), L861X (X being any amino acid), L858R, E746K, L747S, E749Q, A750P, A755V, V765M, C797S, L858P or T790M substitution, deletion of E746-A750, deletion of R748-P753, insertion of Ala (A) between M766 and A767, insertion of Ser, Vai and Ala (SVA) between S768 and V769, insertion of Asn and Ser (NS) between P772 and H773, insertion of one or more amino acids between D761 and E762, A763 and Y764, Y764 and Y765, M766 and A767, A767 and V768, S768 and V769, V769 and D770, D770 and N771, N771 and P772, P772 and H773, H773 and V774, V774 and C775, one or more deletions in EGFR exon 20, or one or more insertions in EGFR exon 20, or any combination thereof.

88. The method of claim 87, wherein the mutant KRAS comprises a G12V, G12C or G12A substitution.

89. The method of any one of claims 75-88, wherein the method inceases immune cell infiltration into a solid tumor, further wherein the immune cells are selected from the group consisting of T cells, B cells, and natural killer cells.

90. The method of claim 89, wherein the T cells comprise CD8+ T cells and CD4+ T cells.

91. The method of claim 90, wherein the T cells comprise CD8+ T cells.

92. The method of any one of claims 75-91, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

93. The method of any one of claims 75-92, wherein the subject is suspected to have or has an EGFR, c-Met or EGFR and c-Met expressing cancer.

94. The method of claim 75, wherein the subject is resistant or has acquired resistance to treatment with a prior anti-cancer therapy.

95. The method of claim 94, wherein the prior anti-cancer therapy is a kinase inhibitor.

96. The method of claim 95, wherein the kinase inhibitor is an inhibitor of EGFR, c-Met, HER2, HER3, HER4, VEGFR or AXL.

97. The method of claim 96, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

98. The method of any one of claims 75-97, wherein the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

99. The method of any one of claims 75-98, wherein the cancer is resistant to treatment with PD-(L)1 axis inhibitors.

100. The method of any one of claims 75-99, wherein the cancer is selected from the group consisting of head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), or non-small cell lung cancer (NSCLC).

101. The method of any one of claims 75-100, further comprising administering one or more anti-cancer therapies to the subject.

102. The method of claim 101, wherein the one or more anti-cancer therapies comprise a kinase inhibitor.

103. The method of claim 102, wherein the kinase inhibitor is an inhibitor of EGFR, c- Met, HER2, HER3, HER4, VEGFR or AXL.

104. The method of claim 103, wherein the kinase inhibitor is erlotinib, gefitinib, lapatinib, vandetanib, afatinib, osimertinib, lazertinib, poziotinib, criotinib, cabozantinib, capmatinib, axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib or sunitinib.

105. The method of claim 94, wherein the prior anti-cancer therapy is chemotherapy.

106. The method of claim 94, wherein the prior anti-cancer therapy is a targeted anti-cancer therapy.

107. The method of claim 101, wherein the one or more anti-cancer therapies comprise chemotherapy.

108. The method of claim 101, wherein the one or more anti-cancer therapies comprise a targeted anti-cancer therapy.

109. The method of any one of claims 75-108, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are administered essentially concurrently to at two different injection sites.

110. The method of any one of claims 75-108, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L)1 axis inhibitor are combined prior to administration and administered simultaneously.

111. The method of any one of claims 75-108, wherein the bispecific anti- EGFR/c-Met antibody and the PD-(L) 1 axis inhibitor are administered sequentially over the course of one or more days as part of a multi-step treatment regimen.

112. A method of treating a solid tumor, ameliorating cancer progression, enhancing immune cell infiltration into a solid tumor, or reducing lactate production in the tumor microenvironment in a subject in need thereof, the method comprising administering a bispecific anti-EGFR/c-Met antibody to the subject, wherein the subject has received a prior administration of the PD-(L)1 axis inhibitor.

113. The method of claim 112, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

114. The method of claim 113, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

115. The method of claim 113, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

116. The method of claim 113, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

117. The method of claim 112, wherein the prior-administered PD-(L)1 axis inhibitor is an inhibitory antibody.

118. The method of claim 117, wherein the prior-administered inhibitory antibody is selected from the group consisting of cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB- A317, TSR-042, and SHR-1210.

119. The method of any one of claims 112-118, wherein the immune cells are selected from the group consisting of T cells, B cells, and natural killer cells.

120. The method of claim 119, wherein the immune cells comprise CD8⁺ T cells and CD4⁺ T cells.

121. The method of claim 120, wherein the immune cells comprise CD8⁺ T cells.

122. The method of any one of claims 112-120, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

123. A method of reducing lactate production in the tumor microenvironment in a subject in need thereof, the method comprising administering a combination of inhibitory agents to the subject, wherein the combination of inhibitory agents is selected from the group consisting of:

(a) a combination comprising an inhibitor of EGFR, an inhibitor of c-Met and a PD-(L)1 axis inhibitor; and

(b) a combination comprising an inhibitor of EGFR and an inhibitor of c- Met, wherein the subject has received a prior administration of a PD-(L) 1 axis inhibitor.

124. The method of claim 123, wherein the combination of an inhibitor of EGFR and an inhibitor of c-Met comprises a bispecific anti-EGFR/c-Met antibody.

125. The method of claim 124, wherein the bispecific anti-EGFR/c-Met antibody comprises

(a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

126. The method of claim 125, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ ID NO: 16.

127. The method of claim 124, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

128. The method of claim 124, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

129. The method of claim 124, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

130. The method of claim 129, wherein the inhibitory antibody is selected from the group consisting of cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

131. The method of any one of claims 123-130, wherein the population of central memory cytotoxic T cells is increased within the microenvironment of the tumor.

132. The method of any one of claims 123-131, wherein the cancer is an epithelial cell cancer, breast cancer, ovarian cancer, lung cancer, non-small cell lung cancer (NSCLC), lung adenocarcinoma, small cell lung cancer, colorectal cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, head and neck cancer, pharynx cancer, cancer of the nose, pancreatic cancer, skin cancer, oral cancer, cancer of the tongue, esophageal cancer, vaginal cancer, cervical cancer, cancer of the spleen, testicular cancer, gastric cancer, cancer of the thymus, colon cancer, thyroid cancer, liver cancer, hepatocellular carcinoma (HCC) or sporadic or hereditary papillary renal cell carcinoma (PRCC).

133. The method of any one of claims 123-132, wherein the cancer is resistant to treatment with PD-(L)1 axis inhibitors.

134. The method of any one of claims 123-133, wherein the cancer is selected from the group consisting of head and neck squamous cell carcinoma (HNSCC), lung squamous cell carcinoma (LUSC), or non-small cell lung cancer (NSCLC).

135. A kit comprising a first pharmaceutical composition comprising a bispecific anti-EGFR/c-Met antibody and a second pharmaceutical composition comprising a PD-(L)1 axis inhibitor in two or more containers.

136. The kit of claim 135, wherein the bispecific anti-EGFR/c-Met antibody comprises (a) a first domain that binds EGFR, wherein the first domain comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 1, a HCDR2 of SEQ ID NO: 2, a HCDR3 of SEQ ID NO: 3, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 4, a LCDR2 of SEQ ID NO: 5 and a LCDR3 of SEQ ID NO: 6; and

(b) a second domain that binds c-Met, wherein the second domain comprises a HCDR1 of SEQ ID NO: 7, a HCDR2 of SEQ ID NO: 8, a HCDR3 of SEQ ID NO: 9, a LCDR1 of SEQ ID NO: 10, a LCDR2 of SEQ ID NO: 11 and a LCDR3 of SEQ ID NO: 12.

137. The kit of claim 135, wherein

(a) the first domain that binds EGFR comprises a heavy chain variable region (VH) of SEQ ID NO: 13 and a light chain variable region (VL) of SEQ ID NO: 14; and

(b) the second domain that binds c-Met comprises a VH of SEQ ID NO: 15 and a VL of SEQ IDNO: 16.

138. The kit of claim 135, wherein the bispecific anti-EGFR/c-Met antibody is an IgGl isotype.

139. The kit of claim 135, wherein the bispecific anti-EGFR/c-Met antibody comprises a first heavy chain (HC1) of SEQ ID NO: 17, a first light chain (LC1) of SEQ ID NO: 18, a second heavy chain (HC2) of SEQ ID NO: 19 and a second light chain (LC2) of SEQ ID NO: 20.

140. The kit of claim 135, wherein the PD-(L)1 axis inhibitor is an inhibitory antibody.

141. The kit of claim 140, wherein the inhibitory antibody is selected from the group consisting of cetrelimab, pembrolizumab (KEYTRUDA®), nivolumab (OPDIVO®), sintilimab, cemiplimab (LIBTAYO®), tripolibamab, tislelizumab, spartalizumab, camrelizumab, dostralimab, genolimzumab, envafolimab, atezolizumab (TECENTRIQ®), durvalumab (IMFINZI®), avelumab (BAVENCIO®), REGN2810, pidilizumab, MEDI0680, PDR001, PF-06801591, BGB-A317, TSR-042, and SHR-1210.

142. The kit of claim 141, wherein the cetrelimab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 31, a HCDR2 of SEQ ID NO: 32, a HCDR3 of SEQ ID NO: 33, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 34, a LCDR2 of SEQ ID NO: 35 and a LCDR3 of SEQ ID NO: 36.

143. The kit of claim 141, wherein the cetrelimab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 37, and a light chain variable region (VL) of SEQ ID NO: 38; or a heavy chain (HC) of SEQ ID NO: 39, and a light chain (LC) of SEQ ID NO: 40.

144. The kit of claim 141, wherein the pembrolizumab antibody comprises a heavy chain complementarity determining region 1 (HCDR1) of SEQ ID NO: 21, a HCDR2 of SEQ ID NO: 22, a HCDR3 of SEQ ID NO: 23, a light chain complementarity determining region 1 (LCDR1) of SEQ ID NO: 24, a LCDR2 of SEQ ID NO: 25 and a LCDR3 of SEQ ID NO: 26.

145. The kit of claim 141, wherein the pembrolizumab antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 27, and a light chain variable region (VL) of SEQ ID NO: 28; or a heavy chain (HC) of SEQ ID NO: 29, and a light chain (LC) of SEQ ID NO: 30.

146. The kit of any one of claims 135-145, wherein the first pharmaceutical composition comprising the bispecific anti-EGFR/c-Met antibody further comprises a first pharmaceutically acceptable excipient and wherein the second pharmaceutical composition comprising the PD-(L)1 axis inhibitor further comprises a second pharmaceutically acceptable excipient.