Methods for predicting patient response to immunotherapy - Patents.com

Abstract

Disclosed are methods for treating a patient having a solid tumor, the methods comprising: (a) obtaining a tumor sample from the patient; (b) evaluating the sample for the level of a biomarker for at least one of innate and adaptive immune cells; and (c) administering to the patient an effective amount of immunotherapy if the sample contains elevated levels of CD68+PD-L1+ macrophages and CD8+ T cells relative to a control and / or elevated levels of CD20+ B cells relative to a control and / or low levels of CD163 expression and 50% or greater (≧) PD-L1 expression relative to a control.

Description

[Technical field]

[0001] The present disclosure relates generally to methods of treating patients with cancer. [Background technology]

[0002] Novel anticancer therapies, such as immunotherapy, may improve overall survival (OS) in patients suffering from late-stage cancers, such as non-small cell lung cancer (NSCLC). However, improved prognosis and sustained response are not uniformly observed in patients treated with immunotherapy. This discrepancy in positive results has been observed in various cancer types treated with different immunotherapy, such as, for example, patients with advanced NSCLC treated with anti-PD-L1 immunotherapy.

[0003] Therefore, it is of great therapeutic interest to be able to identify patients with a greater probability of experiencing a therapeutic benefit in response to immunotherapy, allowing better treatment decisions based on the clinical status of a particular patient. There is a great need for both predictive and prognostic measures that allow early identification of cancer patients who are likely to respond well to immunotherapy. Early identification of such patients is very important, so that the administration of immunotherapy treatment can be prioritized instead of existing standard of care (SOC) chemotherapy regimens. And to the extent that such positive predictive and prognostic characteristics can be understood and potentially reproduced in individuals lacking these characteristics, it may be possible to significantly improve the overall survival (OS) of even more patients.

[0004] The efficacy of any given cancer treatment is greatly influenced by the intrinsic properties of the tumor and the tumor microenvironment (TME). The TME is the environment generated by the tumor, which contains normal and cancerous cells, surrounding vasculature, infiltrating immune cells, extracellular matrix, and numerous signaling molecules. The TME is at least partially defined by its interaction with surrounding tissues, and can significantly affect the efficacy of any anti-cancer treatment.

[0005] Although it is known that the TME can help determine the efficacy of cancer treatment, there is a lack of information on which factors can reliably correlate with a good response to immunotherapy. This is due in part to the difficulty in correlating TME status with clinically meaningful outcomes. Therefore, there is a need for novel clinically meaningful assessments of TME status to determine predictive and prognostic characteristics that allow reliable prediction of the efficacy of immunotherapy to guide treatment decisions for individual cancer patients to improve OS. Summary of the Invention [Means for solving the problem]

[0006] As described herein, in a first aspect, the disclosure provides a method of treating a patient having a solid tumor, the method comprising: a) obtaining a tumor sample from the patient, b) evaluating the sample for a level of a biomarker for at least one of innate and adaptive immune cells, and c) administering to the patient an effective amount of an immunotherapy if the sample comprises elevated levels of CD68+PD-L1+ macrophages and CD8+ T cells compared to a control and / or elevated levels of CD20+ B cells compared to a control.

[0007] In one embodiment of the first aspect, the patient has non-small cell lung cancer (NSCLC). In one embodiment of the first aspect, the patient has advanced NSCLC. In one embodiment of the first aspect, the tumor sample is obtained from a biopsied or resected tumor. In one embodiment of the first aspect, the biomarkers include one or more of PD-L1, PD-1, CD8, CD68, Ki67, AE1, AE3, CD20, NKp46, FOXP3, ICOS, CD66b, CD1c, and CTLA-4. In one embodiment of the first aspect, the levels of the biomarkers are assessed by immunohistochemistry (IHC) and / or multiplex immunofluorescence (mIF). In one embodiment of the first aspect, administration of the immunotherapy improves overall survival (OS) in the patient compared to patients with tumors without elevated levels of CD68+PDL1+ macrophages and CD8+ T cells compared to controls.

[0008] In one embodiment of the first aspect, the method further comprises (d) administering to the patient a standard of care anti-cancer therapeutic if the sample does not contain elevated levels of CD68+PDL1+ macrophages and CD8+ T cells compared to the control or elevated levels of CD20+ B cells compared to the control; and / or (e) administering to the patient one or more chemokines, cytokines, antibodies, antigen presenting cells, and / or synthetic scaffolds to promote formation of intratumoral tertiary lymphoid tissue-like structures.

[0009] In one embodiment, the standard of care anticancer therapeutics include one or more of cisplatin, gemcitabine, methotrexate, vinblastine, doxorubicin, cisplatin (MVAC), carboplatin, taxanes, temozolomide, dacarbazine, vinflunine, docetaxel, paclitaxel, nab-paclitaxel, vemurafenib, erlotinib, afatinib, cetuximab, bevacizumab, gefitinib, and pemetrexed.

[0010] In one embodiment of the first aspect or its embodiments, the immunotherapy comprises an immune checkpoint inhibitor. In one embodiment, the immune checkpoint inhibitor is one or more of an anti-CTLA-4 antibody, an anti-PD-1 antibody, and an anti-PD-L1 antibody. In one embodiment, the anti-CTLA-4 antibody is tremelimumab or ipilimumab. In one embodiment, the anti-PD-1 antibody is REGN2810, SHR1210, IBI308, PDR001, nivolumab, pembrolizumab, BGB-A317, BCD-100, or JS001. In one embodiment, the anti-PD-L1 antibody comprises durvalumab, avelumab, atezolizumab, KNO35, or sugemalimab. In one embodiment, the anti-PD-L1 antibody comprises durvalumab, avelumab, atezolizumab, or KNO35. In one embodiment, the anti-PD-L1 antibody is durvalumab. In one embodiment, the patient is administered durvalumab at a dose of 10 mg / kg once every two weeks (Q2W).In one embodiment, the patient is administered durvalumab at a dose of 1500 mg once every four weeks (Q4W).

[0011] In one embodiment of the first aspect or embodiments thereof, at least one of cancer cell division, tumor growth, tumor size, tumor density, or tumor metastasis is reduced in the patient.

[0012] In a second aspect, the disclosure provides a method of improving overall survival in a patient having a solid tumor, the method comprising: a) obtaining a tumor sample from the patient, b) evaluating the sample for a level of a biomarker for at least one of innate and adaptive immune cells, and c) administering to the patient an effective amount of an immunotherapy if the sample comprises elevated levels of CD68+PDL1+ macrophages and CD8+ T cells compared to a control and / or elevated levels of CD20+ B cells compared to a control.

[0013] In one embodiment of the second aspect, the patient has NSCLC. In one embodiment of the second aspect, the patient has progressive NSCLC. In one embodiment of the second aspect or its embodiments, the patient is administered durvalumab at a dose of 10 mg / kg Q2W. In one embodiment of the second aspect or its embodiments, the patient is administered durvalumab at a dose of 1500 mg Q4W. In another embodiment of the second aspect or its embodiments, the patient is administered durvalumab at a dose of 1500 mg Q3W.

[0014] In a third aspect, the disclosure provides a method of treating a patient having a solid tumor, the method comprising: (a) obtaining a tumor sample from the patient; (b) evaluating the sample for a level of a biomarker for at least one of innate and adaptive immune cells; and (c) administering an effective amount of an immunotherapy to the patient if the sample comprises low CD163 expression and 50% or more (≧) PD-L1 expression compared to a control. In one embodiment, the low level of CD163 expression is 30% or less (≦). In another embodiment, the sample further comprises a high level of CD45 expression compared to a control.

[0015] In a fourth aspect, the disclosure provides a method of improving overall survival in a patient having a solid tumor, the method comprising: (a) obtaining a tumor sample from the patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) administering to the patient an effective amount of an immunotherapy if the sample comprises a low level of CD163 expression relative to a control, and optionally, an elevated level of CD45 expression relative to a control. [Brief description of the drawings]

[0016] [Figure 1] Relative differences in T effector, B cell, and dendritic cell gene expression profiles between long and short OS groups. [Diagram 2] Relative differences in CD8, CD8 subpopulation, and CD68 subpopulation biomarker protein expression profiles between long and short OS groups. Increased CD8+, CD8+PD1+Ki67+, CD68+PD-L1+, and CD68+PD-L1+Ki67+ population densities correlate with long OS. [Diagram 3] Relative differences in B cell biomarker protein expression profiles between long and short OS groups. Increased density of B cells, indicative of TLS, correlates with longer OS. [Figure 4]Baseline tumors of long OS of NSCLC with TLS (IHC and mIF1a). Figure 4 shows a true immunological battlefield with immune cells fighting tumor cells in tumors of patients with long OS of NSCLC. Top left photo: Light microscopic IHC image showing CD20 staining. High density CD20+ B cells correlate with TLS+ tumors. Enriched TLS+ in 50% of pts with long OS >2yr. Middle photo - Immunofluorescence staining (from mF1a panel) with DAPI (dark blue), CD8 (light blue), PD-L1 (green), PD-L1 (yellow), CD68 (orange), and Ki67 (pink) staining. The elongated nuclear shape in the proliferation (Ki67 pink) is likely a follicular dendritic cell (FDC). Top right photo - Enlarged panel of the middle photo indicated by dashed line. Arrow A - CD68+ cell (likely antigen presenting cell (APC)) in association with PD1+ cell (likely T helper); arrow B - CD8+ cytotoxic T cell in association with tumor cell; arrow C - CD8+Ki67+, proliferating cell. Bottom right photo - CD68+ macrophage phagocytosing a neoplastic cell (arrow D). Bottom left photo - enlarged panel of the middle photo indicated by dashed line. Arrow E - phagocytic, CD68+ cell with unclear / broken staining near TLC. [Diagram 5] High density of TLS (CD20+ B cells) predicts OS benefit (median overall survival (mOS) not reached (NR)) compared with 16.6 months in patients with PD-L1 ≥ 25% treated with durvalumab for NSCLC. Twenty-five percent of patients (9 of 35) showed high CD20+ cell density, which correlated with mOS NR. [Figure 6] Relative measure of innate immune response in the TME based on macrophage density. CD68+PD-L1+ and CD68+PD-L1+Ki67+ macrophage density was significantly higher in patients with long OS (n=19 for each) compared to patients with short OS (N=62 and 60, respectively). [Figure 7]Immunofluorescence image of a long OS patient tumor sample stained for mIF1a panel. The dashed circle highlights a CD68+ (orange) PD-L1+ (green) macrophage surrounding / in association with the tumor cell while in association with a CD8 (light blue) T cell. [Figure 8] Relative measure of adaptive immune response in the TME based on CD8 T cell density. CD8+ and CD8+PD-1+Ki67+ T cell densities were significantly higher in patients with long OS (n=36 and 17, respectively) compared to patients with short OS (N=129 and 59, respectively). [Figure 9] Combination of immune markers using optimized cut-offs for OS. Patients with both biomarkers: high CD8+ T cells and high macrophage CD68+PD-L1+ show a significant increase in median OS (39.5 months) compared to single high biomarkers (16.6-20.2 months). These observations may provide good predictive value for immunotherapy. Curve A - high CD68+PD-L1+ and high CD8+; Curve B - PD-L1>=25%; Curve C - PD-L1<25%; Curve D - high CD68+PD-L1+ and low CD8+; Curve E - low CD68+PD-L1+ and low CD8+; Curve F - low CD68+PD-L1+ and high CD8+. PD-L1 TC status reported as %PD-L1 TC, SP263 Ab Ventana central laboratory. PD-L1 density is derived from mIF: digitally measured as cells / mm2; using SP263 Ab. Biomarker profile correlates with mOS and HR (95% CI). [Figure 10] High density of CD68+PD-L1+ macrophages and CD8+ T cells predicts mOS of 39.5 months compared to mOS of 16.6 months in PD-L1≧25% NSCLC patients treated with durvalumab. mOS based on high marker density or combined markers (relative to PD-L1 TC status) based on optimized marker cutoffs for patient prevalence. CD20 is not shown as high mOS for CD20+ is NR. [Figure 11]Computational illustration for cutoff optimization - trade-off between morbidity vs. mOS / mPFS. TME: tumor microenvironment; NSCLC: non-small cell lung cancer; mOS: median overall survival; mPFS: median progression-free survival; ORR: overall response rate; HR: hazard ratio; mIF: multiplex immunofluorescence assay; Ph3: phase 3; TLS: tertiary lymphoid tissue-like structures; NR: not reached. [Figure 12-1] High CD68+PD-L1+ or high CD8+ was associated with OS benefit with improved mOS and HR when compared with single biomarker:PD-L1 TC ≥ 25% (mOS 16.6; HR 0.55 (95% CI: 0.51-0.60)).TME: tumor microenvironment; NSCLC: non-small cell lung cancer; mOS: median overall survival; mPFS: median progression-free survival; ORR: overall response rate; HR: hazard ratio; mIF: multiplex immunofluorescence assay; Ph3: phase 3; TLS: tertiary lymphoid tissue-like structures; NR: not reached. [Figure 12-2] Same as above. [Figure 13-1] Overall survival in NSCLC patients treated with durvalumab and analyzed by CD163, STAB1 (Figure 13A) or C1QC protein (Figure 13B) expression (M2 markers) and PD-L1 status (50% cut-off). Figure 13A: Curve A - STAB1_low; PD-L1>=50%; Curve B - STAB1_high; PD-L1>=50% Curve C - STAB1_low; PD-L1<50% Curve D - STAB1_high; PD-L1<50% Figure 13B: Curve A - C1QC_low; PD-L1>=50%; Curve B - C1QC_high; PD-L1>=50% Curve C - C1QC_low; PD-L1<50% Curve D - C1QC_high; PD-L1<50%. High CD163 protein expression and other M2-associated markers (STAB1, C1QC) were associated with inadequate durvalumab benefit, even in patients with PD-L1 TC ≥ 50% NSCLC. [Figure 13-2] Same as above. [Figure 13-3] Same as above. [Figure 13-4] Same as above. [Figure 14-1]The mOS benefit from immune infiltration at baseline occurs only in subjects with low CD163 expression. Figure 14A: Curve A - CD163_low; PTPRC_high Curve B - CD163_high; PTPRC_high; Curve C - CD163_low; PTPRC_low; Curve D - CD163_high; PTPRC_low. High immune infiltration (PTPRC=CD45) is associated with OS benefit. High CD45 and high CD163 are associated with poor OS benefit, while high CD45 and low CD163 expression are associated with OS benefit. Furthermore, patients with poorer prognosis have significantly higher baseline CD163 protein expression as a percentage of overall CD45 protein expression (Figure 14A). [Figure 14-2] Same as above. [Figure 14-3] Same as above. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Unless otherwise defined, all scientific and technical terms used herein have the meanings commonly understood by those skilled in the art to which this disclosure belongs. The following references provide those skilled in the art with general definitions of many of the terms used in this disclosure: Singleton, et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger, et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). The following terms used herein have the meanings ascribed to them unless otherwise specified.

[0018] The terms "administration" and "administering" as used herein refer to providing, contacting, and / or delivering immune therapy (or immunotherapy) or other therapeutic agents (such as chemotherapeutic agents) by any suitable route to achieve a desired effect in a subject. Examples of administration of therapeutic agents may include, but are not limited to, oral, sublingual, parenteral (e.g., intravenous, subcutaneous, intradermal, intramuscular, intra-articular, intra-arterial, intra-synovial, intrasternal, intrathecal, intralesional, or intracranial injection), transdermal, topical, buccal, rectal, vaginal, nasal, and / or ophthalmic administration. Administration of therapeutic agents may also be via inhalation. A further form of administration of therapeutic agents may be via an indwelling agent, in which a reservoir of therapeutic agent is introduced into a subject and the therapeutic agent is released from the reservoir in clinically meaningful concentrations over a predefined period, such as a week or more, months, or years.

[0019] The term "co-administered" or "administered in combination" as used herein refers to the simultaneous or sequential administration of multiple therapeutic compounds or agents. A first therapeutic compound or agent may be administered prior to, simultaneously with, or after administration of a second therapeutic compound or agent. A first therapeutic compound or agent and a second therapeutic compound or agent may be administered simultaneously or sequentially on the same day, or sequentially within 1, 2, 3, 4, 5, 6 days, 1, 2, 3 weeks, or 1 month of each other. In some embodiments, the therapeutic compounds or agents are co-administered while each of the therapeutic compounds or agents exerts at least some physiological effect and / or remains effective.

[0020] As used herein, a "therapeutic agent" or "therapeutic compound" may refer to a substance, such as an antibody, chemical, and / or pharmaceutical composition, that when administered in a therapeutically effective amount to a subject in need thereof, provides a therapeutic benefit to a subject having the particular disease or disorder being treated. As used herein, a "therapeutic benefit" refers to the eradication or amelioration of the underlying disease being treated and / or the eradication or amelioration of one or more of the symptoms associated with the underlying disease, such that a subject being treated with the therapeutic agent reports an improvement in feeling or condition, although the subject may still be afflicted by the underlying disease.

[0021] Therapeutic agents contemplated for use herein include immune checkpoint inhibitors, chemotherapeutic agents, and radiation therapy. Examples of immune checkpoint inhibitors include anti-PD-L1 antibodies and / or anti-CTLA4 antibodies. Other examples are described herein.

[0022] "Anti-PD-L1 antibody" refers to an antibody that selectively binds a PD-L1 polypeptide. Exemplary anti-PD-L1 antibodies are described, for example, in U.S. Patent Nos. 8,779,108, 9,493,565; and 10,400,039, which are incorporated by reference for all purposes. Durvalumab (MEDI4736) or "Durva" is an exemplary anti-PD-L1 antibody suitable for the methods described herein. Other anti-PD-L1 antibodies may also be used.

[0023] "Anti-CTLA4 antibody" refers to an antibody that selectively binds to CTLA4 polypeptide.Exemplary anti-CTLA4 antibodies are described in, for example, U.S. Patent Nos. 6,682,736; 7,109,003; 7,132,281; 7,411,057; 7,824,679; 8,143,379; 7,807,797; and 8,491,895 (wherein tremelimumab is 11.2.1), which are incorporated herein by reference for all purposes.Ipilimumab and tremelimumab, or "Treme", are exemplary anti-CTLA4 antibodies.Other anti-CTLA4 antibodies can also be used.

[0024] The terms "biomarker" and "marker" (which may be used interchangeably), as used herein, generally refer to proteins, nucleic acid molecules, clinical indicators, and / or other analytes associated with a cell type. In one embodiment, a biomarker may be differentially expressed (or present) in a biological sample obtained from a subject with a disease (e.g., lung cancer) relative to the concentration present in a control sample or reference. In further embodiments, a biomarker may include a measure of gene expression of a particular gene of interest.

[0025] In another embodiment, the biomarker can be an indicator of the density of a cell type within a tissue sample. For example, when the concentration of a biomarker is elevated in a patient with a disease (e.g., cancer) compared to a control (e.g., a subject without cancer), the elevated concentration can indicate the presence of a particular cell type, e.g., an immune cell, and can indicate the presence of an immune response associated with the disease.

[0026] In some embodiments, the concentration of one or more biomarkers or the density of cells expressing such biomarkers can be indicative of a patient's immune fitness, e.g., the relative ability of a patient's immune system to fight a particular disease by itself or to be enhanced or modified by treatment with a therapeutic agent, such as an immune checkpoint inhibitor (ICI), to fight a particular disease, such as cancer.

[0027] In this disclosure, the terms "comprise," "including," "containing," "having," and the like, can have the meaning given them in United States patent law and can mean "comprise," "including," and the like; the terms "consisting essentially of" and "consisting essentially of" likewise have the meaning given them in United States patent law, and the terms are open-ended and permit the presence of more than what is recited, but exclude prior art embodiments, so long as the basic or novel characteristics of what is recited are not altered by the presence of more than what is recited.

[0028] As used herein, the terms "determining," "assessing," "assaying," "measuring," "detecting," and "identifying" refer to both quantitative and qualitative determinations, and therefore these terms may be used interchangeably. Where a quantitative determination is intended, the phrase "determining the amount" of an analyte, substance, protein, etc. may be used. Where a quantitative determination is intended, the phrase "detecting an analyte" may be used.

[0029] The term "disease" refers to any condition or disorder that damages, impedes, or dysregulates the normal function of a cell, tissue, or organ. In diseases such as cancer (e.g., lung cancer), the normal function of a cell, tissue, or organ can be altered to allow immune evasion and / or immune escape of cancer cells or tumors.

[0030] The terms "immune therapy," "immunotherapy," and "immunotherapy" refer to the treatment of disease by activating or suppressing the immune system. Activation immunotherapy amplifies the immune response, while suppression immunotherapy reduces or suppresses the immune response.

[0031] "Subject" or "patient" means a mammal, including, but not limited to, a human, such as a human patient, a non-human primate, or a non-human mammal, such as a bovine, equine, canine, ovine, or feline.

[0032] As used herein, the terms "treat," "treating," "treatment," and the like refer to reducing, alleviating, diminishing, ameliorating, arresting, or ameliorating a disorder and / or symptoms associated therewith. Although not excluded, it will be recognized that treatment of a disorder or condition does not require the complete disappearance of the disorder, condition, or symptoms associated therewith.

[0033] Unless otherwise stated or clear from the context, the term "or" as used herein is understood to be inclusive. Unless otherwise stated or clear from the context, the terms "a," "an," and "the" as used herein are understood to be singular or plural. Similarly, when a particular term is expressed in the singular, the same term expressed in the plural is also intended, and vice versa. For example, the term "drug" also contemplates "drugs," and vice versa.

[0034] Unless otherwise stated or clear from the context, as used herein, the term "about" is understood to refer to within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. All numerical values ​​provided herein are modified by the term about unless clear from the context.

[0035] overview The present disclosure is directed to methods of treating patients with cancer, such as advanced non-small cell lung cancer (NSCLC), to obtain better overall survival (OS) based on improved understanding of predictive and prognostic features that correlate with better immunotherapy efficacy. The present disclosure provides a significant advancement in cancer patient treatment methodology, as it allows clinicians to make better treatment decisions for treating cancer.

[0036] The types of cancer or "solid tumors" contemplated for treatment herein include, for example, NSCLC, advanced solid malignancies, bile duct tumors, bladder cancer, colorectal cancer, diffuse large B-cell lymphoma, esophageal tumors, esophageal squamous cell carcinoma, extensive stage small cell lung cancer, gastric adenocarcinoma, gastric cancer, gastroesophageal junction cancer, head and neck cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma, Hodgkin's lymphoma, lung cancer, melanoma, mesothelioma, metastatic clear cell renal carcinoma, metastatic melanoma, metastatic noncutaneous melanoma, multiple myeloma, nasopharyngeal carcinoma, non-Hodgkin's lymphoma, ovarian cancer, fallopian tube cancer, peritoneal tumors, pleural mesothelioma, prostate tumors, recurrent or metastatic PD-L1 positive or negative SCCHN, recurrent squamous cell lung cancer, renal cell cancer, renal cell carcinoma ... carcinoma, SCCHN, pharyngohypophyseal squamous cell carcinoma, laryngeal squamous cell carcinoma, small cell lung cancer, squamous cell carcinoma of the head and neck, squamous cell lung cancer, TNBC, transitional cell carcinoma, unresectable or metastatic melanoma, urothelial cancer, and urothelial carcinoma.

[0037] As described herein, the disclosure features, in one embodiment, a method of treating a patient having cancer, such as small cell lung cancer (NSCLC), by a) obtaining a tumor sample from the patient, b) evaluating the sample for a level of a biomarker for at least one of innate immune cells and adaptive immune cells, and c) administering to the patient an effective amount of an immunotherapy, if the sample contains at least one of an elevated level of an innate immune cell biomarker or an elevated level of an adaptive immune cell biomarker.

[0038] In one embodiment, an immune checkpoint inhibitor is provided for use in treating a patient with a solid tumor, wherein the sample of the solid tumor comprises an increased level of CD68+PDL1+ macrophages and CD8+ T cells compared to a control; and / or an increased level of CD20+ B cells compared to a control. In another embodiment, an anti-PDL1 antibody is provided for use in treating a patient with a solid tumor, wherein the sample of the solid tumor comprises an increased level of CD68+PDL1+ macrophages and CD8+ T cells compared to a control; and / or an increased level of CD20+ B cells compared to a control. In another embodiment, durvalumab is provided for use in treating a patient with a solid tumor, wherein the sample of the solid tumor comprises an increased level of CD68+PDL1+ macrophages and CD8+ T cells compared to a control; and / or an increased level of CD20+ B cells compared to a control. In one embodiment, the solid tumor is NSCLC. In a further embodiment, the solid tumor is advanced NSCLC.

[0039] In one embodiment, an immune checkpoint inhibitor is provided for use in treating a patient with a solid tumor, wherein the sample from the solid tumor comprises a low level of CD163 expression and 50% or more PD-L1 expression compared to a control. In another embodiment, an anti-PDL1 antibody is provided for use in treating a patient with a solid tumor, wherein the sample from the solid tumor comprises a low level of CD163 expression and 50% or more PD-L1 expression compared to a control. In another embodiment, durvalumab is provided for use in treating a patient with a solid tumor, wherein the sample from the solid tumor comprises a low level of CD163 expression and 50% or more PD-L1 expression compared to a control. In one embodiment, the low level of CD163 expression is 30% or less. In another embodiment, the sample further comprises a high level of CD45 expression compared to a control. In one embodiment, the solid tumor is NSCLC. In another embodiment, the solid tumor is advanced NSCLC.

[0040] In one embodiment, a method for diagnosing a patient with a solid tumor as a candidate for treatment with an immunotherapy to achieve improved overall survival is provided. The method includes (a) obtaining a tumor sample from the patient, (b) evaluating the sample for the level of a biomarker for at least one of innate and adaptive immune cells, and (c) diagnosing the patient as a candidate for treatment with an immunotherapy (e.g., an immune checkpoint inhibitor) if the sample contains elevated levels of CD68+PD-L1+ macrophages and CD8+ T cells compared to a control and / or elevated levels of CD20+ B cells compared to a control. The immune checkpoint inhibitor can be any of anti-CTLA-4 antibodies, anti-PD-1 antibodies, and anti-PD-L1 antibodies as disclosed elsewhere herein. For example, the immune checkpoint inhibitor can be durvalumab, which can be administered at a dose of 10 mg / kg once every two weeks (Q2W), 1500 mg once every four weeks (Q4W), or 1500 mg once every three weeks (Q3W).

[0041] In some embodiments, the patient has advanced NSCLC. In some embodiments, the tumor sample may be obtained from a tumor biopsy or a tumor that has been totally or partially resected. Biomarkers that may be assessed in the tumor sample may include one or more of PD-L1, PD-1, CD8, CD68, Ki67, AE1, AE3, CD20, NKp46, FOXP3, ICOS, CD66b, CD1c, and CTLA-4. Other biomarkers include CD163 (a marker of suppressive (M2) macrophages) and CD45. In some embodiments, the degree or level of expression of the biomarker may be assessed at the transcriptional level by measuring gene expression. In other embodiments, biomarker expression may be assessed at the protein level by immunohistochemistry (IHC) and / or multiplex immunofluorescence (mIF). In other embodiments, biomarker expression may be assessed by proteomic mass spectrometry. Other techniques for measuring biomarker protein expression are contemplated herein as known in the art.

[0042] Additionally, biomarkers contemplated herein may include, but are not limited to, T (CD3+), B (CD19+), NK (CD56+), T naive CD4+ and CD8+ (CD3+CD4 / CD8+CD45RA+CD45RO-CCR7+), Treg / activated (ACT; CD3+CD4+CD25hi / bright CD127low / -), TEM CD4+ (CD3+CD4+CD45RA-CD45RO+CCR7-), TCM CD4+ and CD8+ (CD3+CD4+ / CD8+CD45RA-CD45RO+CCR7+), and T CD3+CD4+ICOS+ and CD4+CD38+ cells, in addition to those indicative of M-MDSC, granulocytes, monocytes, and neutrophils.

[0043] In further embodiments, contemplated biomarkers may include changes in gene expression of specific genes of interest, which may include, but are not limited to, T effector genes, natural killer (NK) cell genes, B cell genes, and dendritic cell (DC) genes. Examples include gene expression signatures for T effector-associated genes (CD8A, EOMES, GZMA, GZMB, CXCL9, CXCL10, IFNG, TBX21), NK cell-associated genes (NCR1 (NKp48), GNLY, KLRC3, KLRD1, KLRF1, NCR1), B cell-associated genes (CD19, MS4A1, CD22, CD79A), neutrophil-associated genes (CD177, and DC-associated genes (CD1C, KIT, CCR7, BATF3, FLT3, ZBTB46, IRF8, BTLA, and MYCL). Additional biomarkers may include ARG1, IL10, HLA-DRA, and HLA-DRB1.

[0044] Measurement of gene and / or protein biomarker expression in tumors can provide a general assessment of the relative presence of adaptive and / or innate immune cells in the tumor, and can provide useful insight into how a patient may respond to immunotherapy, such as treatment with immune checkpoint inhibitors. Indeed, without wishing to be bound by theory, it is believed that patients with one or more tumors that have at least one of elevated levels of innate immune cell biomarkers or elevated levels of adaptive immune cell biomarkers will experience better overall survival (OS) when treated with immunotherapy, such as durvalumab, compared to patients that do not have elevated levels of such biomarkers. Furthermore, it is believed that elevated levels of adaptive and / or innate immune cell biomarkers in tumors can indicate the presence of tertiary lymphoid tissue-like structures (TLS) associated with the tumor (or within the TME), which may play an important role in patients experiencing improved OS.

[0045] Tertiary lymphoid structures (TLS) are ectopic lymphoid organs that develop in non-lymphoid tissues at sites of chronic inflammation, such as tumors. TLS can mature in tumors to promote adaptive anti-tumor immune responses that translate to clinical benefit in patients with cancer. Promotion of intratumoral TLS formation in patients lacking intratumoral TLS (e.g., via treating patients with chemokines, cytokines, antibodies, antigen-presenting cells and / or synthetic scaffolds) may result in improved outcomes for immunotherapy treatment. (Sautes-Fridman et al.Tertiary lymphoid structures in the era of cancer immunotherapy.Nat Rev Cancer 19,307-325(2019)).

[0046] Thus, if a patient is found not to exhibit an active anti-tumor immune response and / or elevated levels of adaptive and / or innate immune cell biomarkers indicative of the presence of TLS, a treatment decision may be made to administer a standard of care anti-cancer therapeutic to the patient. As contemplated herein, standard of care anti-cancer therapeutics may include one or more of cisplatin, gemcitabine, methotrexate, vinblastine, doxorubicin, cisplatin (MVAC), carboplatin, taxanes, temozolomide, dacarbazine, vinflunine, docetaxel, paclitaxel, nab-paclitaxel, vemurafenib, erlotinib, afatinib, cetuximab, bevacizumab, gefitinib, and pemetrexed. Further examples include drugs that target the DNA damage repair system, such as poly(ADP-ribose) polymerase 1 (PARP1) inhibitors and therapeutic agents that inhibit WEE1 protein kinase activity, ATR protein kinase activity, ATM protein kinase activity, Aurora B protein kinase activity, and DNA-PK activity.

[0047] Furthermore, administration of one or more chemokines, cytokines, antibodies, antigen-presenting cells and / or synthetic scaffolds to patients to promote intratumoral TLS formation may be necessary to improve (or indeed enable) subsequent treatment with immunotherapies such as durvalumab to improve OS. Examples of contemplated TLS-promoting compounds are described, for example, in Sautes-Fridman et al. Tertiary Lymphoid Structures in Cancers: Prognostic Value, Regulation, and Manipulation for Therapeutic Intervention. Front Immunol. 7:407 (2016).

[0048] In some embodiments, contemplated immunotherapies may include immune checkpoint inhibitors. Examples of immune checkpoint inhibitors include anti-CTLA-4 antibodies, anti-PD-1 antibodies, and anti-PD-L1 antibodies. In some embodiments, the anti-CTLA-4 antibody may be tremelimumab or ipilimumab. In some embodiments, the anti-PD-1 antibody may be REGN2810, SHR1210, IBI308, PDR001, nivolumab, pembrolizumab, BGB-A317, BCD-100, or JS001. In some embodiments, the anti-PD-L1 antibody includes durvalumab, avelumab, atezolizumab, KNO35, or sugemalimab. In some embodiments, the anti-PD-L1 antibody includes durvalumab, avelumab, atezolizumab, or KNO35. In some embodiments, the anti-PD-L1 antibody is durvalumab. Any therapeutically effective subsections of antibodies, such as antigen-binding fragments, are also contemplated herein.

[0049] In some embodiments, the treatment contemplated herein stops, reduces, slows, or otherwise alleviates or improves one or more symptoms of cancer in a patient.For example, the disclosed methods can slow the rate of cancer cell division or tumor growth, reduce tumor size or tumor density, and / or slow or stop tumor metastasis in a patient.In some embodiments, the treatment improves OS.

[0050] Additionally, the disclosure features, in another embodiment, a method of improving overall survival in patients with advanced NSCLC. The method includes a) obtaining a tumor sample from the patient, b) evaluating the sample for a level of a biomarker for at least one of innate and adaptive immune cells, and c) administering an effective amount of an immunotherapy to the patient if the sample contains elevated levels of CD68+PDL1+ macrophages and CD8+ T cells compared to a control. In one embodiment, a combination of high density of CD68+PDL1+ macrophages and CD8+ T cells is associated with improved or longer OS compared to any single biomarker.

[0051] In a further embodiment, the disclosure features a method of improving overall survival in patients with advanced NSCLC. The method includes a) obtaining a tumor sample from the patient, b) evaluating the sample for a level of a biomarker for at least one of innate and adaptive immune cells, and c) administering an effective amount of an immunotherapy to the patient if the sample contains an elevated level of CD20+ B cells. An elevated level of CD20+ B cells indicates the presence of tertiary lymphoid tissue-like structures (TLS) associated with the tumor. Furthermore, a high density of CD20+ B cells in the sample indicates the presence of TLS and is associated with a long OS.

[0052] Additionally, the disclosure features, in another embodiment, a method of improving overall survival in a patient with advanced NSCLC, the method comprising: a) obtaining a tumor sample from the patient; (b) evaluating the sample for a level of a biomarker for at least one of innate and adaptive immune cells; and (c) administering to the patient an effective amount of an immunotherapy if the sample comprises a low level of CD163 expression relative to a control, and optionally an elevated level of CD45 expression relative to a control.

[0053] In some embodiments, the immunotherapy may be administered at one or more doses of about 1, or about 3, or about 10, or about 15 mg / kg every 1, 2, 3, or 4 weeks. For example, a patient may be treated with durvalumab at a dose of 10 mg / kg Q2W.

[0054] Further, the disclosure features, in another embodiment, a use of an immunotherapy for the manufacture of a medicament for treating a patient having a solid tumor, the use including: (a) obtaining a tumor sample from the patient; (b) evaluating the sample for a level of a biomarker for at least one of innate immune cells and adaptive immune cells; and (c) administering to the patient an effective amount of the immunotherapy if the sample includes elevated levels of CD68+PD-L1+ macrophages and CD8+ T cells relative to a control and / or elevated levels of CD20+ B cells relative to a control.

[0055] In another embodiment, the disclosure provides a use of an immunotherapy for the manufacture of a medicament for treating a patient having a solid tumor, the use comprising: (a) obtaining a tumor sample from the patient; (b) evaluating the sample for a level of a biomarker for at least one of innate immune cells and adaptive immune cells; and (c) administering to the patient an effective amount of the immunotherapy if the sample comprises a low level of CD163 expression and greater than or equal to 50% PD-L1 expression relative to a control.

[0056] The amount of anti-cancer therapeutic, such as an antibody or antigen-binding fragment thereof, to be administered to a patient will depend on various parameters, such as the patient's age, weight, clinical evaluation, immunocompatibility, TME status, tumor burden, and / or other factors including the judgment of the attending physician. Any acceptable route of administration is contemplated, including, but not limited to, intravenous (e.g., intravenous infusion), parenteral, or subcutaneous routes of administration.

[0057] Any therapeutic composition or method contemplated herein can be combined with one or more of any of the other therapeutic compositions and methods provided herein.

[0058] In some embodiments, the treatment regimen may include a biological component, such as an antibody and one or more of a TLS-promoting component and a chemotherapeutic component.

[0059] In some embodiments, kits are contemplated that include an antibody and a biological component, such as one or more of a TLS-promoting component and a chemotherapeutic component. EXAMPLES

[0060] The following examples illustrate certain embodiments of the present invention and various uses thereof. They are set forth for illustrative purposes only and should not be construed as limiting the scope of the disclosure in any way.

[0061] Example 1 - Presence of tertiary lymphoid tissue-like structures (TLS) and combined high density of PD-L1+ macrophages and CD8+ T cells predicts long-term overall survival (OS) for durvalumab-treated patients with advanced non-small cell lung cancer (NSCLC) overview Baseline PD-L1 expression has demonstrated its clinical utility in predicting OS in NSCLC patients receiving anti-PD-(L)1 therapy, including durvalumab. However, beyond the tumor PD-L1-T cell axis, the impact of the tumor microenvironment (TME) and immune composition (e.g., the relative number of immune cell types present) in light of the interplay between innate and adaptive cells deserves to be explored in the quest for better predictors of durvalumab overall survival for patient selection. To that end, we sought to determine biomarkers of TME / immune composition associated with short OS of less than 1 year versus long OS of more than 2 years in NSCLC patients treated with durvalumab using RNA sequencing (RNAseq). As a result of the RNAseq analysis, we developed multiplex immunofluorescence (mIF1a / b) and IHC to explore the overall immune composition, including tertiary lymphoid tissue-like structures (TLS).

[0062] Better pre-existing adaptive (T, B cell) and innate (MAC, DC and NK) immunity, including TLS, shows a strong association with durvalumab-treated NSCLC patients having long OS (>2 years) versus short OS (<1 year). Fifty percent of patients with long OS of NSCLC with high density of CD20+ cells show the presence of TLS with increased APC-T-cell and immune cell-tumor pairings. In the vicinity of TLS, an active anti-tumor immune response is observed via phagocytosis. These findings highlight the important interplay between innate-adaptive immune cells and immune-tumor cells and suggest its relevance for the presence of TLS.

[0063] introduction Predictive biomarkers for anti-PD-(L)1 therapy (e.g., durvalumab) have mostly focused on the tumor-T cell axis, and tumor cell PD-L1 expression has demonstrated its clinical utility in predicting overall survival (OS) in patients with advanced non-small cell lung cancer (NSCLC). Although other immune cell subsets have been shown to be associated with clinical efficacy, their relative influence and combined effect in predicting improved long-term survival warrants further investigation. Using computational image analysis of multiplex immunofluorescence (mIF) and immunohistochemistry (IHC) immune marker panels, we sought to identify single and combined biomarkers of tumor immune composition associated with long-term OS in patients with advanced NSCLC treated with durvalumab.

[0064] method Pretreatment tumor samples from patients (n=210) with advanced NSCLC enrolled in a nonrandomized Phase 1 / 2 trial of durvalumab (10mg / kg Q2W, CP1108 / NCT01693562) were analyzed via RNAseq to identify candidate biomarkers of TME / immune composition associated with long versus short OS.

[0065] Tumor samples were then stained using IHC and a 6-marker mIF panel developed as a result of the RNAseq analysis to detect markers of immune cells, cell functional status, and tertiary lymphoid tissue-like structures (TLS). The panel is shown in Table No. 1. Cell marker density (cells / mm 2 ), distribution, and proximity were quantified and analyzed in relation to OS.

[0066] [Table 1]

[0067] result Significant differences in T effector, B cell, and dendritic cell gene expression profiles were observed between the long and short OS groups (Figure 1). As shown in Figures 2-12, computational image analysis of tumor immune architecture revealed superior immune inflammatory phenotypes, both innate (macrophages, dendritic cells) and adaptive (T and B cells), in NSCLC patients with long OS compared to those with short OS (fold change >2, p<0.0001). Subgroups of patients with a high density of individual immune subsets exhibit a median OS (mOS) of 10-20 months (p<0.01 high vs. low subgroups), while combined markers of innate and adaptive immune cells show improved mOS >2 years (p<0.001).

[0068] Specifically, among the key findings, the combined biomarker of high density of CD68+PD-L1+ macrophages and high density of CD8+ T cells was significantly superior to the single biomarker, CD68+PD-L1+ macrophages (20.2 months mOS, p<10 -6 , HR=0.28, 95% CI 0.17~0.48) or CD8+ T cells (18.4 months mOS, p<10 -7 , HR=0.39, 95% CI 0.27-0.55), predicting a significant increase in mOS of 39.5 months (p-value <10 -7 , HR=0.21, 95%CI 0.12-0.39). Furthermore, a high density of CD20+ cells, reflecting B-cell tumor infiltration and the presence of TLS, predicted long-term OS (mOS NR, p=0.003). Tumors enriched for TLS exhibit increased levels of macrophages expressing PD-L1 in association with CD8+ T cells (activated PD1+ or proliferative Ki67+ T cells).

[0069] Consideration Our findings demonstrate the importance of both tertiary lymphoid tissue-like structures and high pre-existing innate-adaptive immunity in promoting long-term overall survival in patients with NSCLC treated with durvalumab and highlight the need for the development of multiparameter predictive biomarkers outside the tumor-T-cell axis.

[0070] Example 2 - The functional status of tumor-associated macrophages influences the clinical outcome of durvalumab in patients with NSCLC as revealed by proteomic mass spectrometry introduction In Example 1, we use computational image analysis of multiplex immunofluorescence (mIF) to show the positive impact of CD68+PD-L1+ macrophages in combination with CD8+ T cells in predicting long-term OS benefit in NSCLC patients treated with durvalumab (anti-PD-L1), highlighting the influence of the myeloid compartment on the IO response.Using, in part, proteomic mass spectrometry, we sought to investigate further functional impact of myeloid cells on the IO response and, in particular, on suppressive (M2) tumor-associated macrophages (TAMs).

[0071] method Pretreatment tumor samples from 66 patients with advanced NSCLC enrolled in a nonrandomized Phase 1 / 2 trial of durvalumab (10 mg / kg Q2W, CP1108 / NCT01693562) were processed for global and targeted proteomics. Pathologists determined tumor area and single FFPE tumor tissue sections were used for laser capture macrodissection and protein extraction followed by mass spectrometry using label-free, data-independent and parallel reaction monitoring.

[0072] result Among the immune-related proteins detected by proteomics mass spectrometry, we evaluated CD163 protein expression, a detailed marker of inhibitory (M2) macrophages, and its impact on durvalumab clinical prognosis alone and in association with PD-L1 and immune infiltration. While the proteomics-based biomarker evaluable population (BEP) showed a shorter median (m) OS compared to the intended-to-treat population, we verified that high expression of PD-L1, as measured by IHC or proteomics, was associated with a better OS benefit as predicted when treating with durvalumab. On this basis, evaluation of proteins associated with inhibitory M2 macrophages revealed that approximately 30% of NSCLC patients with high CD163 protein expression showed an inadequate OS benefit (mOS of 5 months) after durvalumab treatment. High CD163 expression and other M2-related markers (STAB1, C1QC) are associated with poor durvalumab benefit, even in PD-L1 TC≧50% NSCLC patients (see FIG. 13). In contrast, patients with PD-L1 TC≧50% and low CD163 expression obtain a superior OS benefit (mOS of 13.4 months) than those with high CD163 expression (OS of 5 months) or PD-L1 TC<50% despite the level of CD163 expression (mOS of 4.1-7.4 months). Furthermore, patients with a poorer prognosis have significantly higher baseline CD163 protein expression as a percentage of overall CD45 protein expression.

[0073] conclusion This analysis further confirms the importance of myeloid cell function within the tumor microenvironment (TME) in determining prognosis for T cell-specific IO therapy and highlights the utility of proteomic mass spectrometry in assessing TME findings more broadly and quantitatively based on RNAseq and IHC / mIF assays.

[0074] All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each individual patent and publication was specifically and individually indicated to be incorporated by reference. Citation or identification of any reference in any section of this application should not be construed as an admission that such reference is available as prior art to the present disclosure.

Claims

1. 1. An immunotherapy for use in a method of treating a patient having a solid tumor, said method comprising: (a) obtaining a tumor sample from said patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) administering to said patient an effective amount of said immunotherapy if said sample contains elevated levels of CD68+PD-L1+ macrophages and CD8+ T cells relative to a control and / or elevated levels of CD20+ B cells relative to a control.

2. The immunotherapy for use according to claim 1, wherein the patient has non-small cell lung cancer (NSCLC).

3. The immunotherapy for use according to claim 2, wherein the patient has advanced NSCLC.

4. The immunotherapy method according to any one of claims 1 to 3, wherein the tumor sample is obtained from a biopsy or a resected tumor.

5. The immunotherapy method for use according to any one of claims 1 to 4, wherein said levels of biomarkers are assessed by immunohistochemistry (IHC) and / or multiplex immunofluorescence (mIF).

6. 6. The immunotherapy for use according to any one of claims 1 to 5, wherein said administration of said immunotherapy improves overall survival (OS) in said patient compared to patients having tumors without elevated levels of CD68+PDL1+ macrophages and CD8+ T cells compared to controls.

7. (d) administering to the patient a standard of care anti-cancer therapeutic if the sample does not contain elevated levels of CD68+ PDL1+ macrophages and CD8+ T cells relative to a control or elevated levels of CD20+ B cells relative to a control; and / or (e) administering to said patient one or more chemokines, cytokines, antibodies, antigen-presenting cells, and / or synthetic scaffolds to promote the formation of intratumoral tertiary lymphoid tissue-like structures. The immunotherapy method according to any one of claims 1 to 6, further comprising:

8. 1. An immunotherapy for use in a method of treating a patient having a solid tumor, said method comprising: (a) obtaining a tumor sample from said patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) administering to said patient an effective amount of said immunotherapy if said sample contains a low level of CD163 expression and 50% or greater (≧) PD-L1 expression relative to a control.

9. The immunotherapy method according to claim 8, wherein the low level of CD163 expression is less than or equal to 30% (≦).

10. The immunotherapy method of claim 8 or claim 9, wherein the sample further comprises an elevated level of CD45 expression compared to a control.

11. The immunotherapy method for use according to any one of claims 8 to 10, wherein the patient has non-small cell lung cancer (NSCLC).

12. The immunotherapy method of claim 11, wherein the patient has advanced NSCLC.

13. The immunotherapy method for use according to any one of claims 8 to 12, wherein said tumor sample is obtained from a biopsy or resected tumor.

14. The immunotherapy for use according to any one of claims 8 to 13, wherein said levels of biomarkers are assessed by immunohistochemistry (IHC) and / or multiplex immunofluorescence (mIF) and / or proteomic mass spectrometry.

15. The immunotherapy for use according to any one of claims 8 to 14, wherein said administration of said immunotherapy improves overall survival (OS) in said patient compared to patients having a tumor with a high level of CD163 protein expression compared to a control.

16. The immunotherapy method of claim 15, wherein said high level of CD163 protein expression is greater than 30%.

17. (d) administering a standard of care anti-cancer therapeutic to the patient if the sample contains a high level of CD163 expression compared to a control; and / or (e) administering to said patient one or more chemokines, cytokines, antibodies, antigen-presenting cells, and / or synthetic scaffolds to promote the formation of intratumoral tertiary lymphoid tissue-like structures. The immunotherapy method according to any one of claims 8 to 16, further comprising:

18. 18. The immunotherapy for use of claim 7 or claim 17, wherein the standard of care anti-cancer therapeutic comprises one or more of cisplatin, gemcitabine, methotrexate, vinblastine, doxorubicin, cisplatin (MVAC), carboplatin, taxanes, temozolomide, dacarbazine, vinflunine, docetaxel, paclitaxel, nab-paclitaxel, vemurafenib, erlotinib, afatinib, cetuximab, bevacizumab, gefitinib, and pemetrexed.

19. The immunotherapy for use according to any one of claims 1 to 18, wherein the immunotherapy comprises an immune checkpoint inhibitor.

20. The immunotherapy method for use according to claim 19, wherein the immune checkpoint inhibitor is one or more of an anti-CTLA-4 antibody, an anti-PD-1 antibody and an anti-PD-L1 antibody.

21. The immunotherapy for use according to claim 20, wherein the anti-CTLA-4 antibody is tremelimumab or ipilimumab.

22. The immunotherapy for use according to claim 20, wherein the anti-PD-1 antibody is REGN2810, SHR1210, IBI308, PDR001, nivolumab, pembrolizumab, BGB-A317, BCD-100, or JS001.

23. The immunotherapy method of claim 20, wherein the anti-PD-L1 antibody is durvalumab, avelumab, atezolizumab, KNO35 or sugemalimab.

24. The immunotherapy method of claim 23, wherein the anti-PD-L1 antibody is durvalumab.

25. 25. The immunotherapy for use according to claim 24, wherein the patient is administered durvalumab at a dose of 10 mg / kg once every two weeks (Q2W).

26. 25. The immunotherapy for use according to claim 24, wherein the patient is administered durvalumab at a dose of 1500 mg once every four weeks (Q4W).

27. 25. The immunotherapy for use according to claim 24, wherein the patient is administered durvalumab at a dose of 1500 mg once every three weeks (Q3W).

28. The immunotherapy for use according to any one of claims 1 to 27, wherein at least one of cancer cell division, tumor growth, tumor size, tumor density, or tumor metastasis is reduced in said patient.

29. 1. A method of treating a patient having a solid tumor, comprising: (a) obtaining a tumor sample from said patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) if the sample contains elevated levels of CD68+PD-L1+ macrophages and CD8+ T cells relative to a control and / or elevated levels of CD20+ B cells relative to a control, administering to the patient an effective amount of an immunotherapy. A method comprising:

30. 30. The method of claim 29, wherein the patient has non-small cell lung cancer (NSCLC).

31. 31. The method of claim 30, wherein the patient has advanced NSCLC.

32. The method of any one of claims 29 to 31, wherein the tumor sample is obtained from a biopsy or resected tumor.

33. The method of any one of claims 29 to 32, wherein the levels of biomarkers are assessed by immunohistochemistry (IHC) and / or multiplex immunofluorescence (mIF).

34. 34. The method of any one of claims 29-33, wherein said administration of said immunotherapy improves overall survival (OS) in said patient compared to patients having tumors without elevated levels of CD68+ PDL1+ macrophages and CD8+ T cells compared to controls.

35. (d) administering to the patient a standard of care anti-cancer therapeutic if the sample does not contain elevated levels of CD68+ PDL1+ macrophages and CD8+ T cells relative to a control or elevated levels of CD20+ B cells relative to a control; and / or (e) administering to said patient one or more chemokines, cytokines, antibodies, antigen-presenting cells, and / or synthetic scaffolds to promote the formation of intratumoral tertiary lymphoid tissue-like structures. The method of any one of claims 29 to 34, further comprising:

36. 1. A method of treating a patient having a solid tumor, comprising: (a) obtaining a tumor sample from said patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) if the sample contains a low level of CD163 expression and 50% or greater (≧) PD-L1 expression relative to a control, administering to the patient an effective amount of an immunotherapy. A method comprising:

37. The method of claim 36, wherein the low level of CD163 expression is less than or equal to 30% (≦).

38. 38. The method of claim 36 or claim 37, wherein the sample further comprises an elevated level of CD45 expression compared to a control.

39. The method of any one of claims 36 to 38, wherein the patient has non-small cell lung cancer (NSCLC).

40. 40. The method of claim 39, wherein the patient has advanced NSCLC.

41. The method of any one of claims 36 to 40, wherein the tumor sample is obtained from a biopsy or resected tumor.

42. The method of any one of claims 36 to 41, wherein said levels of biomarkers are assessed by immunohistochemistry (IHC) and / or multiplex immunofluorescence (mIF) and / or proteomic mass spectrometry.

43. 43. The method of any one of claims 36-42, wherein said administration of said immunotherapy improves overall survival (OS) in said patient compared to patients having tumors with high levels of CD163 expression compared to controls.

44. The method of claim 43, wherein the high level of CD163 expression is greater than 30%.

45. (d) administering a standard of care anti-cancer therapeutic to the patient if the sample contains a high level of CD163 expression compared to a control; and / or (e) administering to said patient one or more chemokines, cytokines, antibodies, antigen-presenting cells, and / or synthetic scaffolds to promote the formation of intratumoral tertiary lymphoid tissue-like structures. The method of any one of claims 36 to 44, further comprising:

46. 46. ​​The method of claim 35 or claim 45, wherein the standard of care anti-cancer therapeutic comprises one or more of cisplatin, gemcitabine, methotrexate, vinblastine, doxorubicin, cisplatin (MVAC), carboplatin, taxanes, temozolomide, dacarbazine, vinflunine, docetaxel, paclitaxel, nab-paclitaxel, vemurafenib, erlotinib, afatinib, cetuximab, bevacizumab, gefitinib, and pemetrexed.

47. 47. The method of any one of claims 29 to 46, wherein the immunotherapy comprises an immune checkpoint inhibitor.

48. 48. The method of claim 47, wherein the immune checkpoint inhibitor is one or more of an anti-CTLA-4 antibody, an anti-PD-1 antibody, and an anti-PD-L1 antibody.

49. 49. The method of claim 48, wherein the anti-CTLA-4 antibody is tremelimumab or ipilimumab.

50. 49. The method of claim 48, wherein the anti-PD-1 antibody is REGN2810, SHR1210, IBI308, PDR001, nivolumab, pembrolizumab, BGB-A317, BCD-100, or JS001.

51. 49. The method of claim 48, wherein the anti-PD-L1 antibody comprises durvalumab, avelumab, atezolizumab, KNO35, or sugemalimab.

52. 52. The method of claim 51, wherein the anti-PD-L1 antibody is durvalumab.

53. 53. The method of claim 52, wherein the patient is administered durvalumab at a dose of 10 mg / kg once every two weeks (Q2W).

54. 53. The method of claim 52, wherein the patient is administered durvalumab at a dose of 1500 mg once every four weeks (Q4W).

55. 53. The method of claim 52, wherein the patient is administered durvalumab at a dose of 1500 mg once every three weeks (Q3W).

56. 56. The method of any one of claims 29 to 55, wherein at least one of cancer cell division, tumor growth, tumor size, tumor density, or tumor metastasis is reduced in the patient.

57. 1. A method of improving overall survival in a patient having a solid tumor, comprising: (a) obtaining a tumor sample from said patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) if the sample contains elevated levels of CD68+ PDL1+ macrophages and CD8+ T cells relative to a control and / or elevated levels of CD20+ B cells relative to a control, administering to the patient an effective amount of an immunotherapy. A method comprising:

58. 1. A method of improving overall survival in a patient having a solid tumor, comprising: (a) obtaining a tumor sample from said patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) if the sample contains a low level of CD163 expression relative to a control, and optionally, an elevated level of CD45 expression relative to a control, administering to the patient an effective amount of an immunotherapy. A method comprising:

59. 59. The method of claim 57 or claim 58, wherein the patient has NSCLC.

60. 60. The method of claim 59, wherein the patient has advanced NSCLC.

61. 61. The method of any one of claims 57-60, wherein the patient is administered durvalumab at a dose of 10 mg / kg Q2W.

62. 61. The method of any one of claims 57-60, wherein the patient is administered durvalumab at a fixed dose of 1500 mg Q4W.

63. 61. The method of any one of claims 57-60, wherein the patient is administered durvalumab at a fixed dose of 1500 mg Q3W.

64. Use of an immunotherapy for the manufacture of a medicament for treating a patient with a solid tumor, said use comprising (a) obtaining a tumor sample from said patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) if the sample contains elevated levels of CD68+PD-L1+ macrophages and CD8+ T cells relative to a control and / or elevated levels of CD20+ B cells relative to a control, administering to the patient an effective amount of the immunotherapy. Including, use.

65. Use of an immunotherapy for the manufacture of a medicament for treating a patient with a solid tumor, said use comprising (a) obtaining a tumor sample from said patient; (b) evaluating the sample for a level of a biomarker for at least one of an innate immune cell and an adaptive immune cell; and (c) if the sample contains a low level of CD163 expression and greater than or equal to 50% PD-L1 expression relative to a control, administering to the patient an effective amount of the immunotherapy. Including, use.

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