EP4673161A2

EP4673161A2 - Oncostatin-m-based chimeric antigen receptor (car)-immune cells

Info

Publication number: EP4673161A2
Application number: EP24764714.2A
Authority: EP
Inventors: Reshmi Parameswaran; Daniel Feinberg
Original assignee: Case Western Reserve University
Current assignee: Case Western Reserve University
Priority date: 2023-03-02
Filing date: 2024-03-04
Publication date: 2026-01-07
Also published as: JP2026507180A; CN121152632A; AU2024229562A1; WO2024182803A2; WO2024182803A3

Abstract

A chimeric antigen receptor (CAR) includes an extracellular antigen binding domain, wherein the extracellular antigen binding domain comprises a polypeptide that binds to an oncostatin-M receptor (OSMR), leukemic inhibitory factor receptor (LIFR), a heterodimer between transmembrane glycoprotein 130 (gp130) receptor and OSMR, and/or heterodimer between gp130 receptor and LIFR.

Description

ONCOSTATIN M-BASED CHIMERIC ANTIGEN RECEPTOR (CAR) IMMUNE CELLS

RELATED APPLICATION

[0001] This application claims priority from U.S. Provisional Application No. 63/487,919, filed March 2, 2023, the subject matter of which is incorporated herein by reference in its entirety.

SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on March 4, 2024, is named CWR-032533WO ORD.st.26 and is 58,919 bytes in size.

BACKGROUND

[0003] Chimeric antigen receptor (CAR)-T cell therapy has become a mainstay treatment in the arsenal of doctors treating liquid tumors. FDA approval has been received for both CD 19 and BCM A targeting CAR-T cell therapies. Initial CD 19 CAR-T trials in pediatric B-cell acute lymphoblastic leukemia (ALL) showed 93% achievement of ‘complete remission’ and a relapse rate of 31.5 percent. While CD19 and BCMA CAR-T cell therapies have not been as effective in all B-cell/plasma cell tumors respectively, the degree and duration of response they have been able to achieve in highly relapsed patients is remarkable. Less than one year ago, CD 19 CAR-T cell therapy received FDA approval as a first-line therapy for adults with large B-cell lymphoma, showing that these therapies are not just a last-resort for the highly relapsed/refractory population. Yet, for as successful as CAR-T therapy has been in liquid tumors, the impact in solid tumors has been limited, and no CAR-T therapy has been FDA approved to treat any solid cancers.

[0004] There are two key reasons that CAR-T cells fail in solid tumors, one is the inability to penetrate the tumor microenvironment (TME), and the second is the inability to kill the cancer itself. For example, mesothelin is overexpressed in mesothelioma cells, and mesothelin-targeting-CAR-T cells have been created. Combining the results of the three completed phase I human trials utilizing mesothelin-targeting-CAR-Ts against mesothelioma, only 9 out of 48 patients had any response, and all 9 were transient responses of stable disease. This disappointing response is not limited to just mesothelioma, but many other solid tumor CAR-T trials. SUMMARY

[0005] Embodiments described herein relate to a chimeric antigen receptor (CAR) that includes an extracellular antigen binding domain that targets a receptor of oncostatin-M (OSM), an engineered CAR immune cell that expresses the CAR, and the use of the CAR immune cells in a CAR immunotherapy. The extracellular antigen binding domain of the CAR includes a polypeptide that binds to an oncostatin-M receptor (OSMR), leukemic inhibitory factor receptor (LIFR), a heterodimer between transmembrane glycoprotein 130 (gpl30) receptor and OSMR, and/or a heterodimer between gpl30 receptor and LIFR that is expressed by cancer cells and surrounding cells that support these cancer cells in a tumor microenvironment. The CAR can be expressed from immune cells, such as cytotoxic T lymphocytes (T cells), natural killer (NK) cells, and/or natural killer T (NKT) cells, to target and kill OSMR and/or LIFR expressing cancer cells and surrounding cells that support these cancer cells in a tumor microenvironment.

[0006] In some embodiments, the polypeptide comprises an oncostatin-M (OSM) ligand or fragment thereof that binds to OSMR and/or LIFR and/or heterodimers of OSMR/gpl30 receptor and/or LlFR/gpl30 receptor.

[0007] In some embodiments, the OSM ligand includes a polypeptide having an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 7, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 16, or SEQ ID NO: 18.

[0008] In other embodiments, the polypeptide includes a single chain variable fragment (ScFv) that binds to OSMR and/or LIFR.

[0009] In some embodiments, the CAR further comprises one or more hinge domains.

[0010] In some embodiments, the hinge domain can include a hinge domain of CD8a having an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 11%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 19.

[0011] In other embodiments, the hinge domain can include a hinge domain of IgGl having an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 20.

[0012] In another embodiment, the CAR further comprises one or more transmembrane domains. The transmembrane domain can include, for example, a CD8a transmembrane domain, a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[0013] In some embodiments, the CD8a transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81 %, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 21.

[0014] In some embodiments, the CD28 transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about '1'1%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 22.

[0015] In other embodiments, the CD 16a transmembrane domain can include an amino acid sequence at least about 70%, at least about 71 %, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 23.

[0016] In other embodiments, the 2B4 transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 24.

[0017] In other embodiments, the NKG2D transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 25.

[0018] In another embodiment, the CAR further comprises one or more intracellular signaling domains. The intracellular signaling domain can include, for example, a 41BB intracellular signaling domain, a CD28 intracellular signaling domain, a CD3^ intracellular signaling domain, aCD16A intracellular signaling domain, a yc intracellular signaling domain, a 2B4 intracellular signaling domain, a DAP10 intracellular signaling domain, a DAP12 intracellular signaling domain, and/or combinations thereof.

[0019] In some embodiments, the 4 IBB intracellular signaling domain(s) can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81 %, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 26.

[0020] In some embodiments, the CD28 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 27.

[0021] In some embodiments, the CD3^ intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 28.

[0022] In some embodiments, the 0X40 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 29.

[0023] In some embodiments, the CD16A intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 30.

[0024] In some embodiments, the yc intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 31 .

[0025] In some embodiments, the 2B4 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 32.

[0026] In some embodiments, the DAP 10 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 33.

[0027] In some embodiments, the DAP 12 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 34.

[0028] In some embodiments, the CAR can further include a spacer. The spacer can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 35.

[0029] In another embodiment, the CAR further comprises a signaling peptide.

[0030] In some embodiments, the signaling peptide comprises an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 36 or SEQ ID NO: 37.

[0031] In some embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a hinge domain, a transmembrane domain, a CD28 intracellular domain; and an 0X40 intracellular domain. [0032] In some embodiments, the hinge domain is an IgGl hinge domain, and the transmembrane domain is a CD28 transmembrane domain.

[0033] In other embodiments, the CAR includes, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, an IgGl hinge domain, a CD28 transmembrane domain, the CD28 intracellular domain, the 0X40 intracellular domain, and a CD3-zeta intracellular domain.

[0034] In some embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a CD 16a intracellular signaling domain, a 2B4 intracellular domain, and a CD3 intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[0035] In other embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a yc intracellular signaling domain, a 2B4 intracellular domain, and a CD3^ intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[0036] In other embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a CD28 intracellular signaling domain, a 2B4 intracellular domain, and a CD3^ intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[0037] In other embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a 2B4 intracellular signaling domain, a 41BB intracellular domain, and a CD3 intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[0038] In other embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a 2B4 intracellular signaling domain, a DAP 10 intracellular domain, a DAP 12 intracellular domain, and a CD3 intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[0039] Other embodiments relate to a nucleic acid or nucleotide comprising a nucleotide sequence encoding a CAR described herein. The nucleotide can be operably linked to a promoter and provided in an expression construct. The expression construct can include a vector, such as a retroviral vector, a lentiviral vector, or an AAV vector.

[0040] In some embodiments, the expression construct can further include a nucleotide sequence encoding a cytokine. The cytokine can include, for example, IL- 15, IL- 12, IL-2, IL- 18, IL-21, or a combination thereof.

[0041] In some embodiments, the expression construct can include a nucleotide sequence that encodes the CAR and the cytokine. For example, the nucleotide sequence can be at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 6.

[0042] Still other embodiments relate to an immune cell comprising a CAR as described herein. The immune cell can be transfected or transduced with a vector comprising the expression construct described herein.

[0043] In some embodiments, the immune can include at least one of a T cell, a NK cell, or a NKT cell.

[0044] In some embodiments, the T cells, NK cells or NKT cells are isolated from a human.

[0045] In other embodiments, the T cells, NK cells or NKT are autologous.

[0046] In still other embodiments, the T cells, NK cells or NKT are allogenic.

[0047] Other embodiments relate to an immunotherapy composition comprising a plurality of CAR expressing immune cells as described herein. [0048] Still other embodiments, relate to a method of treating cancer in a subject in need thereof. The method includes administering to the subject a therapeutically effective amount of the immunotherapy composition as described herein.

[0049] In some embodiments, the cancer includes a solid tumor. For example, the cancer can include at least one of osteosarcoma, melanoma, mesothelioma, breast cancer, prostate cancer, gastric cancer, head and neck cancer, lung cancer, or brain cancer.

[0050] In other embodiments, the cancer includes a hematological cancer or malignancy, such as acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, Acute lymphoblastic leukemia, non- Hodgkin’ s lymphoma, Hodgkin’s lymphoma, B cell malignancies, and multiple myeloma.

[0051] In another embodiment, the composition is co-administered with one or more chemotherapeutic agents.

BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figs. l(A-H) illustrate graphs showing OSM CARs express OSM and preferentially kill specific cell lines. A/B - Displays OSM detected on the surface of transduced human CD3+ T-cells from 2 donors. GFP- reflects CAR-Ts that are not expressing CAR- vector with GFP reporter, but that were transduced with CAR- vectorcontaining virus. C - Cell line’s OSMR surface expression shown as ratio vs IgG isotype control signal on flow cytometry. D/E - Cytotoxicity assays utilizing propidium iodide reporter on flow cytometry with 7:1 ratio of CAR vs target cell for 16 hours. F - Cytokine release assay taken from supernatant after 16 hour incubation with listed cell line. Results are shown as median fluorescent intensity and are relative values. G/H - CD107a staining performed to determine extent of T-cell degranulation. Intra (G) is comparison between a CAR transduced well separating CAR-expressing cells (GFP+) vs Untransduced T-cells (GFP-). Inter (H) is between the CAR wells and the Untransduced control wells. UT stands for Untransduced T-cells. Student’s unpaired T-test performed for analysis of two groups. One-way ANOVA with Tukey’s post-hoc test for multiple comparisons utilized for more than two groups. NS - not significant. P values shown in the graph. [0053] Figs. 2(A-C) illustrate an image and graphs showing intratumoral injection of OSM-CARs reduces solid tumor burden. A - AGS-luciferase tagged tumors were subcutaneously injected and allowed to grow until luminescent signal was e8. Subsequently one mouse was injected with 4,000,000 CAR-T cells intratumorally. Seven days post injection mouse radiance values are shown. N=2 for PBS and n=l for CAR. B - Detroit 562 cells were subcutaneously injected and allowed to grow until palpable. Subsequently 4,000,000 OSM CAR-T cells, Untransduced T-cells (UT), or PBS (vehicle) was injected into the tumors. After 12-14 days tumors were extracted and measured. PBS (n=6), UT (n=5), OSM (n=7). One way ANOVA with Tukey’s Post-Hoc test for multiple comparisons was performed. NS = no significance. * = p<.05. ** = p<.01

[0054] Figs. 3(A-E) illustrate a schematic, graph, and plots showing intravenously (IV) injected OSM CAR-T cells display cytotoxicity against subcutaneous SAOS2. A - Schematic of experiment. End point defined as tumor volume over 1700mm³. B - Tumor volume measured via calipers in two dimensions with formula Length x width², 21 days post injection of SAOS2 and 11 days after T-cell injection IV. C - Displays tumor volume over time D - Displays mass of mice 7 days after injection of CAR-T cells. E - Displays Kaplan- Meier curve of survival of mice in days since SAOS2 injection. Student’s unpaired T-Test performed. Kaplan-Meier survival analysis performed. P-values shown as numeric values. [0055] Figs. 4(A-D) illustrate a schematic, charts, and image showing higher dose of OSM CAR-Ts increases Tumor Killing and Toxicity. A - Shows schematic of experiment. B - Shows mass of extracted SAOS2 tumors 5 days after IV injection of 6,000,000 T-cells or vehicle. C - Shows volume of extracted tumors measured as LxWxD 5 days after injection. D - displays photo of extracted tumors. One-way ANOVA performed with Tukey’s post-hoc test for multiple comparisons. P-values shown as numeric values. Red box indicates tumors not included in analysis as due to missed subcutaneous injection, tumors were systemically present in abdomen. L, LL, R, RR, etc. are identifiers of individual mice.

[0056] Figs. 5(A-E) illustrate a schematic and graphs showing intravenously injected OSM CARs display cytotoxicity against subcutaneous 143B. A - Schematic of experiment. B - Toxicity results in reduced mass of mice shortly after injection (mass shown 7 days after injection.) C/D/E - Tumor volumes shown at Day 7, 14, and 20 post injection of T-cells IV. One way ANOVA with Tukey’s post-hoc test for multiple comparisons performed. P-values displayed numerically. X indicates mouse euthanized for loss in mass exceeding IACUC guidelines.

[0057] Figs. 6(A-E) illustrate a schematic, images, plot, and graph showing intravenously injected OSM CARs display insignificant cytotoxicity against subcutaneous Jeko cells. A - Schematic of experiment, Jeko cells were allowed to grow for 14 days prior to injection of T-cells or vehicle. B - Luciferase tagged jeko cells fluorescence was measured prior to injection of T-cells IV (Day 0) and at Day 7 via Spectrum Imaging System. C - Luciferase imaging values (radiance) displayed over time. D - Radiance of Tumors 14 days after T-cell injection. E - Extracted tumors displayed. L, R, NC are identifiers for individual mice. 3 mice were used per group. Student’s unpaired T-test performed. NS = no significance.

DETAILED DESCRIPTION

[0058] Methods involving conventional molecular biology techniques are described herein. Such techniques are generally known in the art and are described in detail in methodology treatises, such as Current Protocols in Molecular Biology, ed. Ausubel et al., Greene Publishing and Wiley-Interscience, New York, 1992 (with periodic updates). Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains. Commonly understood definitions of molecular biology terms can be found in, for example, Rieger et al., Glossary of Genetics: Classical and Molecular, 5th Ed., Springer- Verlag: New York, 1991, and Lewin, Genes V, Oxford University Press: New York, 1994. The definitions provided herein are to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present invention.

[0059] It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a pharmaceutical carrier" includes mixtures of two or more such carriers, and the like. "Optional" or "optionally" means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. [0060] As used herein, the term “about” means that amounts, sizes, formulations, parameters, and other quantities and characteristics are not and need not be exact, but may be approximate and/or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art. When the term “about” is used in describing a value or an endpoint of a range, the disclosure should be understood to include the specific value or endpoint referred to. Whether or not a numerical value or endpoint of a range in the specification recites “about,” the numerical value or endpoint of a range is intended to include two embodiments: one modified by “about,” and one not modified by “about.” It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.

[0061] The term "about" or "approximately" refers to a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length that varies by as much as 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% to a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length. In one embodiment, the term "about" or "approximately" refers a range of quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length + 15%, ± 10%, ± 9%, ± 8%, ± 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, or ± 1% about a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length.

[0062] The terms “substantial,” “substantially,” and variations thereof as used herein are intended to note that a described feature is equal or approximately equal to a value or description. For example, a “substantially planar” surface is intended to denote a surface that is planar or approximately planar. Moreover, “substantially” is intended to denote that two values are equal or approximately equal. In some embodiments, “substantially” may denote values within about 10% of each other, such as within about 5% of each other, or within about 2% of each other.

[0063] It is noted that the terms “substantially” and “about” may be utilized herein to represent the inherent degree of uncertainty that may be attributed to any quantitative comparison, value, measurement, or other representation. These terms are also utilized herein to represent the degree by which a quantitative representation may vary from a stated reference without resulting in a change in the basic function of the subject matter at issue. Thus, cells that are “free of’ or “substantially free of T cell contamination” for example, are cells to which T cells are not actively added or batched into cell culture, but may be present in very small as a contaminant resulting from natural cell progression during expansion. Similarly, other components may be characterized as “free of’ or “substantially free of’ in the same manner. Further, as used herein, the term “consisting essentially of’ allows for elements not explicitly recited but excludes element that affect basic or novel characteristics of the inventions. As recited herein, the term “consisting of’ excludes elements not expressly stated.

[0064] The term “engineered” as used herein refers to an entity that is generated by the hand of man, including a cell, nucleic acid, polypeptide, vector, and so forth. In at least some cases, an engineered entity is synthetic and comprises elements that are not naturally present or configured in the manner in which it is utilized in the disclosure. In specific embodiments, a vector is engineered through recombinant nucleic acid technologies, and a cell is engineered through transfection or transduction of an engineered vector.

[0065] The term "Nucleic acids," "nucleic acid molecules," "nucleotides," "nucleotide(s) sequence," and "polynucleotide" are used interchangeably and refer to the phosphate ester polymeric form of ribonucleosides (adenosine, guanosine, uridine or cytidine; "RNA molecules") or deoxyribonucleosides (deoxyadenosine, deoxy guanosine, deoxythymidine, or deoxycytidine; "DNA molecules"), or any phosphoester analogs thereof, such as phosphorothioates and thioesters, in either single stranded form, or a double- stranded helix. Single stranded nucleic acid sequences refer to single-stranded DNA (ssDNA) or single- stranded RNA (ssRNA). Double stranded DNA-DNA, DNA-RNA and RNA-RNA helices are possible. The term nucleic acid molecule, and in particular DNA or RNA molecule, refers only to the primary and secondary structure of the molecule, and does not limit it to any particular tertiary forms. Thus, this term includes double-stranded DNA found, inter aha, in linear or circular DNA molecules (e.g., restriction fragments), plasmids, supercoiled DNA and chromosomes. In discussing the structure of particular double-stranded DNA molecules, sequences can be described herein according to the normal convention of giving only the sequence in the 5' to 3’ direction along the non-transcribed strand of DNA (i.e., the strand having a sequence homologous to the mRNA). A "recombinant DNA molecule" is a DNA molecule that has undergone a molecular biological manipulation. DNA includes, but is not limited to, cDNA, genomic DNA, plasmid DNA, synthetic DNA, and semi-synthetic DNA. A "nucleic acid composition" of the disclosure comprises one or more nucleic acids as described herein. [0066] As used herein, a "coding region" or "coding sequence" is a portion of polynucleotide which consists of codons translatable into amino acids. Although a "stop codon" (TAG, TGA, or TAA) is typically not translated into an amino acid, it can be considered to be part of a coding region, but any flanking sequences, for example promoters, ribosome binding sites, transcriptional terminators, introns, and the like, are not part of a coding region. The boundaries of a coding region are typically determined by a start codon at the 5' terminus, encoding the amino terminus of the resultant polypeptide, and a translation stop codon at the 3' terminus, encoding the carboxyl terminus of the resulting polypeptide. Two or more coding regions can be present in a single polynucleotide construct, e.g., on a single vector, or in separate polynucleotide constructs, e.g., on separate (different) vectors. It follows, then, that a single vector can contain just a single coding region, or comprise two or more coding regions.

[0067] The term "downstream" refers to a nucleotide sequence that is located 3' to a reference nucleotide sequence. In certain embodiments, downstream nucleotide sequences relate to sequences that follow the starting point of transcription. For example, the translation initiation codon of a gene is located downstream of the start site of transcription.

[0068] The term "upstream" refers to a nucleotide sequence that is located 5' to a reference nucleotide sequence. In certain embodiments, upstream nucleotide sequences relate to sequences that are located on the 5' side of a coding region or starting point of transcription. For example, most promoters are located upstream of the start site of transcription.

[0069] The term "expression" as used herein refers to a process by which a polynucleotide produces a gene product, for example, an RNA or a polypeptide. It includes without limitation transcription of the polynucleotide into messenger RNA (mRNA), transfer RNA (tRNA), small hairpin RNA (shRNA), small interfering RNA (siRNA) or any other RNA product, and the translation of an mRNA into a polypeptide. Expression produces a "gene product." As used herein, a gene product can be either a nucleic acid, e.g., a messenger RNA produced by transcription of a gene, or a polypeptide which is translated from a transcript. Gene products described herein further include nucleic acids with post transcriptional modifications, e.g., polyadenylation or splicing, or polypeptides with post translational modifications, e.g., methylation, glycosylation, the addition of lipids, association with other protein subunits, or proteolytic cleavage. The term "yield," as used herein, refers to the amount of a polypeptide produced by the expression of a gene.

[0070] A "vector" refers to any vehicle for the cloning of and/or transfer of a nucleic acid into a host cell. A vector can be a replicon to which another nucleic acid segment can be attached so as to bring about the replication of the attached segment. A "replicon" refers to any genetic element e.g., plasmid, phage, cosmid, chromosome, virus) that functions as an autonomous unit of replication in vivo, i.e., capable of replication under its own control. The term "vector" includes vehicles for introducing the nucleic acid into a cell in vitro, ex vivo or in vivo. A large number of vectors are known and used in the art including, for example, plasmids, modified eukaryotic viruses, or modified bacterial viruses. Insertion of a polynucleotide into a suitable vector can be accomplished by ligating the appropriate polynucleotide fragments into a chosen vector that has complementary cohesive termini.

[0071] Vectors can be engineered to encode selectable markers or reporters that provide for the selection or identification of cells that have incorporated the vector. Expression of selectable markers or reporters allows identification and/or selection of host cells that incorporate and express other coding regions contained on the vector. Examples of selectable marker genes known and used in the art include: genes providing resistance to ampicillin, streptomycin, gentamycin, kanamycin, hygromycin, sulfonamide, puromycin, and the like; and genes that are used as phenotypic markers, i.e., anthocyanin regulatory genes, isopentanyl transferase gene, and the like. Examples of reporters known and used in the art include: luciferase (Luc), green fluorescent protein (GFP), chloramphenicol acetyltransferase (CAT), P-galactosidase (LacZ), P-glucuronidase (Gus), and the like. Selectable markers can also be considered to be reporters.

[0072] The term "heterologous" means derived from a genotypically distinct entity from that of the rest of the entity to which it is compared or into which it is introduced or incorporated. For example, a polynucleotide introduced by genetic engineering techniques into a different cell type is a heterologous polynucleotide (and, when expressed, can encode a heterologous polypeptide). Similarly, a cellular sequence e.g., a gene or portion thereof) that is incorporated into a viral vector is a heterologous nucleotide sequence with respect to the vector.

[0073] The term "heterologous gene" or “heterologous nucleic acid” refers to a gene that does not naturally occur as part of a viral genome. For instance, a heterologous gene can be a mammalian gene, e.g., a therapeutic gene, e.g., a mammalian gene that encodes a therapeutic protein. In some embodiments, a heterologous gene encodes a protein or portion thereof that is defective or absent in the target cell and/or subject. In some embodiments, the heterologous gene contains one or more exons encoding a protein that is defective or absent in the target cell and/or subject. For example, in some embodiments, the heterologous gene includes one or more trans-splicing molecules, e.g., as described in WO 2017/087900, which is incorporated herein by reference in its entirety. In some embodiments, a heterologous gene includes a therapeutic nucleic acid, such as a therapeutic RNA (e.g., microRNA).

[0074] The term "promoter" refers to a sequence that regulates transcription of a heterologous gene operably linked to the promoter. Promoters provide the sequence sufficient to direct transcription and/or recognition sites for RNA polymerase and other transcription factors required for efficient transcription and can direct cell- specific expression. In addition to the sequence sufficient to direct transcription, a promoter sequence of the invention can also include sequences of other regulatory elements that are involved in modulating transcription (e.g., enhancers, kozak sequences, and introns).

[0075] The term "homology" and "identity" are used synonymously throughout and refer to sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing a position in each sequence, which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base or amino acid, then the molecules are homologous or identical at that position. A degree of homology or identity between sequences is a function of the number of matching or homologous positions shared by the sequences.

[0076] The term "mutant" refers to any change in the genetic material of an organism, in particular a change (i.e., deletion, substitution, addition, or alteration) in a wild type polynucleotide sequence or any change in a wild type protein. The term "variant" is used interchangeably with "mutant". Although it is often assumed that a change in the genetic material results in a change of the function of the protein, the terms "mutant" and "variant" refer to a change in the sequence of a wild type protein regardless of whether that change alters the function of the protein (e.g., increases, decreases, imparts a new function), or whether that change has no effect on the function of the protein (e.g., the mutation or variation is silent). [0077] The term "target cell" refers to any cell that expresses a target gene and which the vector infects or is intended to infect. Vectors can infect target cells that reside in a subject (in situ) or target cells in culture.

[0078] The term "host cell" as used herein refers to, for example microorganisms, yeast cells, insect cells, and mammalian cells, that can be, or have been, used as recipients of ssDNA or vectors. The term includes the progeny of the original cell which has been transduced. Thus, a "host cell" as used herein generally refers to a cell which has been transduced with an exogenous DNA sequence. It is understood that the progeny of a single parental cell may not necessarily be completely identical in morphology or in genomic or total DNA complement to the original parent, due to natural, accidental, or deliberate mutation. In some embodiments, the host cell can be an in vitro host cell.

[0079] The term “subject” generally refers to an individual having a biological sample that is undergoing processing or analysis and, in specific cases, has or is suspected of having cancer. The subject can be any organism or animal subject that is an object of a method or material, including mammals, e.g., humans, laboratory animals (e.g., primates, rats, mice, rabbits), livestock (e.g., cows, sheep, goats, pigs, turkeys, and chickens), household pets (e.g., dogs, cats, and rodents), horses, and transgenic non-human animals. The subject can be a patient, e.g., have or be suspected of having a disease (that may be referred to as a medical condition), such as benign or malignant neoplasia, or cancer. The subject may be undergoing or having undergone treatment. The subject may be asymptomatic. The subject may be healthy individuals but that are desirous of prevention of cancer.

[0080] The terms “treatment” or “treating,” includes any beneficial or desirable effect on the symptoms or pathology of a disease or pathological condition, and may include even minimal reductions in one or more measurable markers of the disease or condition being treated, e.g., cancer. Treatment can involve optionally either the reduction or amelioration of symptoms of the disease or condition, or the delaying of the progression of the disease or condition. “Treatment” does not necessarily indicate complete eradication or cure of the disease or condition, or associated symptoms thereof.

[0081] The term cancer refers to all types of cancer, neoplasm or malignant tumors found in mammals, including leukemias, lymphomas, melanomas, neuroendocrine tumors, carcinomas and sarcomas. The cancer can be of the liquid or solid tumor type. Exemplary cancers that may be treated with a compound, pharmaceutical composition, or method provided herein include lymphoma, sarcoma, bladder cancer, bone cancer, brain tumor, cervical cancer, colon cancer, esophageal cancer, gastric cancer, head and neck cancer, kidney cancer, myeloma, thyroid cancer, leukemia, prostate cancer, breast cancer (e.g., triple negative, ER positive, ER negative, chemotherapy resistant, herceptin resistant, HER2 positive, doxorubicin resistant, tamoxifen resistant, ductal carcinoma, lobular carcinoma, primary, metastatic), ovarian cancer, pancreatic cancer, liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., non-small cell lung carcinoma, squamous cell lung carcinoma, adenocarcinoma, large cell lung carcinoma, small cell lung carcinoma, carcinoid, sarcoma), glioblastoma multiforme, glioma, melanoma, prostate cancer, castration-resistant prostate cancer, breast cancer, triple negative breast cancer, glioblastoma, ovarian cancer, lung cancer, squamous cell carcinoma (e.g., head, neck, or esophagus), colorectal cancer, leukemia, acute myeloid leukemia, lymphoma, B cell lymphoma, or multiple myeloma. Additional examples include, cancer of the thyroid, endocrine system, brain, breast, cervix, colon, head & neck, esophagus, liver, kidney, lung, non-small cell lung, melanoma, mesothelioma, ovary, sarcoma, stomach, uterus or Medulloblastoma, Hodgkin's Disease, Non-Hodgkin's Lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumors, cancer, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, lymphomas, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, endometrial cancer, adrenal cortical cancer, neoplasms of the endocrine or exocrine pancreas, medullary thyroid cancer, medullary thyroid carcinoma, melanoma, colorectal cancer, papillary thyroid cancer, hepatocellular carcinoma, Paget's Disease of the Nipple, Phyllodes Tumors, Lobular Carcinoma, Ductal Carcinoma, cancer of the pancreatic stellate cells, cancer of the hepatic stellate cells, or prostate cancer.

[0082] The term leukemia refers broadly to progressive, malignant diseases of the blood-forming organs and is generally characterized by a distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is generally clinically classified on the basis of (1) the duration and character of the diseaseacute or chronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or monocytic; and (3) the increase or non- increase in the number abnormal cells in the blood-leukemic or aleukemic (subleukemic). Exemplary leukemias that may be treated with a compound, pharmaceutical composition, or method provided herein include, for example, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic leukemia, leukemia cutis, embryonal leukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblastic leukemia, myelocytic leukemia, myeloid granulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasma cell leukemia, multiple myeloma, plasmacytic leukemia, promyelocytic leukemia, Rieder cell leukemia, Schilling’s leukemia, stem cell leukemia, subleukemic leukemia, or undifferentiated cell leukemia. [0083] The terms metastasis, and metastatic cancer can be used interchangeably and refer to the spread of a proliferative disease or disorder, e.g., cancer, from one organ or another non-adjacent organ or body part. Cancer occurs at an originating site, e.g., breast, which site is referred to as a primary tumor, e.g., primary breast cancer. Some cancer cells in the primary tumor or originating site acquire the ability to penetrate and infiltrate surrounding normal tissue in the local area and/or the ability to penetrate the walls of the lymphatic system or vascular system circulating through the system to other sites and tissues in the body. A second clinically detectable tumor formed from cancer cells of a primary tumor is referred to as a metastatic or secondary tumor. When cancer cells metastasize, the metastatic tumor and its cells are presumed to be similar to those of the original tumor. Thus, if lung cancer metastasizes to the breast, the secondary tumor at the site of the breast consists of abnormal lung cells and not abnormal breast cells. The secondary tumor in the breast is referred to a metastatic lung cancer. Thus, the phrase metastatic cancer refers to a disease in which a subject has or had a primary tumor and has one or more secondary tumors. The phrases non-metastatic cancer or subjects with cancer that is not metastatic refers to diseases in which subjects have a primary tumor but not one or more secondary tumors. For example, metastatic lung cancer refers to a disease in a subject with or with a history of a primary lung tumor and with one or more secondary tumors at a second location or multiple locations, e.g., in the breast.

[0084] The term associated or associated with in the context of a substance or substance activity or function associated with a disease (e.g., cancer, such as osteosarcoma, melanoma, mesothelioma, breast cancer, prostate cancer, head and neck cancer, lung cancer, or brain cancer) means that the disease e.g., such as osteosarcoma, melanoma, mesothelioma, breast cancer, prostate cancer, head and neck cancer, lung cancer, or brain cancer) is caused by (in whole or in part), or a symptom of the disease is caused by (in whole or in part) the substance or substance activity or function.

[0085] The term "Chimeric Antigen Receptor" or alternatively a "CAR" refers to a set of polypeptides, typically two in the simplest embodiments, which when in an immune cell, such as a T cell, provides the cell with specificity for a target cell, typically a cancer cell, and with intracellular signal generation. In some embodiments, a CAR comprises at least an extracellular antigen binding domain, a transmembrane domain and a cytoplasmic signaling domain (also referred to herein as "an intracellular signaling domain") comprising a functional signaling domain derived from a stimulatory molecule and/or costimulatory molecule. In some embodiments, the set of polypeptides are in the same polypeptide chain (e.g., comprise a chimeric fusion protein). In some embodiments, the set of polypeptides are not contiguous with each other, e.g., are in different polypeptide chains. In some embodiments, the set of polypeptides include a dimerization switch that, upon the presence of a dimerization molecule, can couple the polypeptides to one another, e.g., can couple an antigen binding domain to an intracellular signaling domain. In one embodiment, the stimulatory molecule of the CAR is the zeta chain associated with the T cell receptor complex. In one aspect, the cytoplasmic signaling domain comprises a primary signaling domain (e.g., a primary signaling domain of CD3-zeta). In one embodiment, the cytoplasmic signaling domain further comprises one or more functional signaling domains of at least one costimulatory molecule as defined below.

[0086] In one embodiment, the CAR comprises a chimeric fusion protein comprising an extracellular antigen binding domain, a transmembrane domain and an intracellular signaling domain comprising a functional signaling domain of a stimulatory molecule. In one embodiment, the CAR comprises a chimeric fusion protein comprising an extracellular antigen binding domain, a transmembrane domain and an intracellular signaling domain comprising a functional signaling domain of a co-stimulatory molecule and a functional signaling domain of a stimulatory molecule. In one embodiment, the CAR comprises a chimeric fusion protein comprising an extracellular antigen binding domain, a transmembrane domain and an intracellular signaling domain comprising two functional signaling domains of one or more co- stimulatory molecule(s) and a functional signaling domain of a stimulatory molecule. Tn one embodiment, the CAR comprises a chimeric fusion protein comprising an extracellular antigen binding domain, a transmembrane domain and an intracellular signaling domain comprising at least two functional signaling domains of one or more co-stimulatory molecule(s) and a functional signaling domain of a stimulatory molecule.

[0087] The terms "T lymphocyte" and "T cell" are used interchangeably and refer to a principal type of white blood cell that completes maturation in the thymus and that has various roles in the immune system, including the identification of specific foreign antigens in the body and the activation and deactivation of other immune cells. A T cell can be any T cell, such as a cultured T cell, e.g., a primary T cell, or a T cell from a cultured T cell line, e.g., Jurkat, SupTl, etc., or a T cell obtained from a mammal. The T cell can be CD3+ cells. The T cell can be any type of T cell and can be of any developmental stage, including but not limited to, CD4+/CD8+ double positive T cells, CD4+ helper T cells (e.g., Thl and Th2 cells), CD8+ T cells (e.g., cytotoxic T cells), peripheral blood mononuclear cells (PBMCs), peripheral blood leukocytes (PBLs), tumor infiltrating lymphocytes (TILs), memory T cells, naive T cells, regulator T cells, gamma delta T cells, and the like. Additional types of helper T cells include cells such as Th3 (Treg), Thl7, Th9, or Tfh cells. Additional types of memory T cells include cells such as central memory T cells (Tcm cells), effector memory T cells (Tern cells and TEMRA cells). The T cell can also refer to a genetically engineered T cell, such as a T cell modified to express a T cell receptor (TCR) or a chimeric antigen receptor (CAR). The T cell can also be differentiated from a stem cell or progenitor cell.

[0088] 'CD4+ T cells" refers to a subset of T cells that express CD4 on their surface and are associated with cell-mediated immune response. They are characterized by the secretion profiles following stimulation, which may include secretion of cytokines such as IFN-y, TNF-a, IE2, IE4 and IE10. "CD4" are 55-kD glycoproteins originally defined as differentiation antigens on T-lymphocytes, but also found on other cells including monocytes/macrophages. CD4 antigens are members of the immunoglobulin supergene family and are implicated as associative recognition elements in MHC (major histocompatibility complex) class Il-restricted immune responses. On T-lymphocytes they define the helper/inducer subset.

[0089] CD8+ T cells" refers to a subset of T cells which express CD8 on their surface, are MHC class I-restricted, and function as cytotoxic T cells. "CD8" molecules are differentiation antigens found on thymocytes and on cytotoxic and suppressor T- lymphocytes. CD8 antigens are members of the immunoglobulin supergene family and are associative recognition elements in major histocompatibility complex class I-restricted interactions.

[0090] The term "NK cell" or "Natural Killer cell" refer to a subset of peripheral blood lymphocytes defined by the expression of CD56 or CD 16 and the absence of the T cell receptor (CD3). As used herein, the terms "adaptive NK cell" and "memory NK cell" are interchangeable and refer to a subset of NK cells that are phenotypically CD3- and CD56+, expressing at least one of NKG2C and CD57, and optionally, CD16, but lack expression of one or more of the following: PLZF, SYK, FcRy, and EAT-2. In some embodiments, isolated subpopulations of CD56+ NK cells comprise expression of CD16, NKG2C, CD57, NKG2D, NCR ligands, NKp30, NKp40, NKp46, activating and inhibitory KIRs, NKG2A and/or DNAM-1. CD56+ can be dim or bright expression.

[0091] The term "NKT cells" or "natural killer T cells" refers to CD Id-restricted T cells, which express a T cell receptor (TCR). Unlike conventional T cells that detect peptide antigens presented by conventional major histocompatibility (MHC) molecules, NKT cells recognize lipid antigens presented by CD Id, a non-classical MHC molecule. Two types of NKT cells are recognized. Invariant or type I NKT cells express a very limited TCR repertoire— a canonical a- chain (Va24-Ial8 in humans) associated with a limited spectrum of P chains (V i 1 in humans). The second population of NKT cells, called non-classical or non-invariant type II NKT cells, display a more heterogeneous TCRa usage. Type I NKT cells are considered suitable for immunotherapy. Adaptive or invariant (type I) NKT cells can be identified with the expression of at least one or more of the following markers, TCR Va24-Jal8, Vbl l, CDld, CD3, CD4, CD8, aGalCer, CD161 and CD56.

[0092] All percentages and ratios used herein, unless otherwise indicated, are by weight. [0093] Embodiments described herein relate to a chimeric antigen receptor (CAR) that includes an extracellular antigen binding domain that targets a receptor of oncostatin-M (OSM), an engineered CAR immune cell that expresses the CAR, and the use of CAR immune cell in CAR immunotherapy. The extracellular antigen binding domain of the CAR includes a polypeptide that binds to an oncostatin-M receptor (OSMR), leukemic inhibitory factor receptor (LTFR), a heterodimer between transmembrane glycoprotein 130 (gpl 30) receptor and OSMR, and/or a heterodimer between gpl 30 receptor and LIFR that is expressed by cancer cells and surrounding cells that support these cancer cells in a tumor microenvironment.

[0094] In some embodiments, the CAR can be expressed from immune cells, such as cytotoxic T lymphocytes (T cells), natural killer (NK) cells, and/or natural killer T (NKT) cells, to target and kill OSMR and/or LIFR expressing cancer cells and surrounding cells that support these cancer cells in the tumor microenvironment. In an example according to the present disclosure, the CAR-T cells can be engineered to target a receptor of OSM using OSM as a ligand. In another example of the present disclosure, natural killer (NK) cells or natural killer T (NKT) cells are modified to express a CAR.

[0095] Oncostatin-M (OSM) is a soluble IL-6 family cytokine that binds to a heterodimer between the transmembrane gpl 30 receptor and either LIFR or OSMR. These heterodimers will be referred to as LIFR and OSMR in future instances. OSM has a higher binding preference to OSMR than LIFR. OSMR is upregulated in many cancers including: lung, triple negative breast, head and neck, and gastric cancers. OSM is secreted by fibroblasts in the tumor microenvironment (TME) and when OSM is knocked out only in surrounding fibroblasts, tumor growth can be greatly reduced. OSM-OSMR binding in these cells that encompass the TME creates a positive feedback loop that supports cancer growth. On the other hand, LIFR is generally downregulated in cancers with rare exceptions that include osteosarcoma. Given OSMR’s upregulation not only in cancer cells themselves, but the surrounding tumor microenvironment, we created an OSM-ligand CAR immune cell that expresses OSM on its surface and found that targeting OSMR via OSM-ligand-CAR-T cell therapy uniquely results in both effective tumor penetration and killing. Thus, OSMR along with LIFR present a target opportunity for treating such cancers.

[0096] Accordingly, one embodiment described herein relates a method of treating or preventing a disease or condition by targeting a receptor of OSM. Another embodiment relates to a method of treating or preventing a disease or condition by targeting a cell expressing or overexpressing a receptor of OSM, such as OSMR or LIFR. In an exemplary embodiment, the disease or condition is a cancer, such as a solid tumor or hematologic malignancy.

[0097] In some embodiments, the solid tumor can be an osteosarcoma, melanoma, mesothelioma, breast cancer, prostate cancer, gastric cancer, head and neck cancer, lung cancer, or brain cancer.

[0098] In other embodiments, the hematologic malignancy can be any hematologic malignancy wherein the cancer cells express or overexpress a receptor of OSM, including but not limited to acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, Acute lymphoblastic leukemia, non-Hodgkin’s lymphoma, Hodgkin’s lymphoma, B cell malignancies, and multiple myeloma.

[0099] In some embodiments, the polypeptide of the extracellular antigen binding domain of the CAR comprises an oncostatin-M (OSM) ligand that binds to OSMR and/or LIFR and/or heterodimers of OSMR/gpl30 receptor and/or LIFR/gpl30 receptor.

[00100] In some embodiments, the “OSM ligand” or “OSM” refers to any of the recombinant or naturally occurring forms of the OSM as set forth in SEQ ID NO: 7 or variants to homologs thereof that maintain OSM activity (e.g., within at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to OSM).

[00101] In some embodiments, the OSM ligand includes a polypeptide having an acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 7.

[00102] In other embodiments, the OSM ligand can be a fragment of the OSM ligand and have a “partial sequence” or “OSM” partial sequence”. A “partial sequence” or “OSM” partial sequence” refers to a portion of SEQ ID NO: 7 that maintains OSM activity similar to that of the whole sequence, and in particular, an extracellular portion of OSM that is responsible for binding with a receptor of OSM. In one example of a partial sequence is a sequence can include at least 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15% or 10% of the naturally occurring OSM sequence. Also contemplated are sequences having at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the partial sequence.

[00103] In other embodiments, the OSM ligand is a mutant OSM ligand that has stronger affinity to LIFR than native or wild-type OSM. Stronger affinity of the mutant OSM ligand is possible through mutations of B-C alpha helical regions. The mutant OSM ligands can have amino acid sequences at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 16, or SEQ ID NO: 18.

[00104] In other embodiments, the polypeptide of the extracellular antigen binding domain of the CAR can include a single chain variable fragment (scFv) that binds to OSMR and/or LIFR. The scFv can include humanized and chimeric antibody fragments or antigen binding fragments thereof that selectively bind to a OSMR and/or LIFR. The antibody fragments and antigen binding fragments thereof can be used in an immunotherapy to treat cancer in humans or other mammals.

[00105] Methods for making humanized scFvs from non-human versions are well known in the art. Humanization is essentially performed by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. The choice of human variable domains, both light and heavy, to be used in making the humanized antibodies can, in some instances, be important to reduce antigenicity and/or human anti-mouse antibody (HAMA) response.

[00106] The variable domain of the antibodies refers to certain portions of the variable domains that differ in sequence among antibodies. The variability in the antibodies and antigen binding fragments thereof can be concentrated in three CDR segments, located in both the light chain and the heavy chain variable domains. The highly conserved portions of variable domains are called framework (FR) regions. In the antibodies described herein, there are four FR regions, connected by three CDRs, that can comprise a variable chain. The CDRs in each of the light and heavy chains are held together in close proximity by the FR regions and, with the CDRs from the other chain, can contribute to the formation of the target binding site of antibodies.

[00107] Antibody humanization is a process that can generate engineered human antibodies with variable region ("V-region") sequences that are substantially similar to actual human germline sequences, while retaining the binding specificity and affinity of a reference antibody. This process can graft, for example, the CDR1, CDR2, and CDR3 regions of the heavy and the light chain sequences into humanized human framework that is both optimized and previously identified prior to the start of the grafting process. The variable region containing the humanized framework can be produced into single chain antigen binding fragments thereof. The resulting engineered humanized antibody fragments can retain the binding specificity of the parent murine antibody for the OSMR or LIFR, and can have an equivalent or higher binding affinity for a specific antigen than the parent antibody. The engineered antigen binding fragments can have heavy and light chain V-regions with a high degree of amino acid sequence identity compared to the closest human germline antibody genes. For example, additional maturational changes can be introduced in the CDR3 regions of each chain during construction in order to identify antibodies with optimal binding kinetics.

[00108] In some embodiments, the CAR can further comprise one or more hinge domains.

[00109] In some embodiments, the hinge domain can include a hinge domain of CD8a having an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 19.

[00110] In other embodiments, the hinge domain can include a hinge domain of IgGl having an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81 %, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 20.

[00111] In yet another example, the CAR cells of the present disclosure comprise a hinge domain of CD28. In yet another example, the CAR cells of the present disclosure comprise a hinge domain of FCyRIlL

[00112] In another embodiment, the CAR can further include one or more transmembrane domains. The transmembrane domain can include, for example, a CD8a transmembrane domain, a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[00113] In some embodiments, the CD8a transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 21.

[00114] In some embodiments, the CD28 transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 22.

[00115] In other embodiments, the CD16a transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about '1'1%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 23.

[00116] In other embodiments, the 2B4 transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 24.

[00117] In other embodiments, the NKG2D transmembrane domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 25. [00118] In another embodiment, the CAR further comprises one or more intracellular signaling domains. The intracellular signaling domain can include, for example, a 41BB intracellular signaling domain, a CD28 intracellular signaling domain, a CD3^ intracellular signaling domain, aCD16A intracellular signaling domain, a yc intracellular signaling domain, a 2B4 intracellular signaling domain, a DAP10 intracellular signaling domain, a DAP12 intracellular signaling domain, and/or combinations thereof.

[00119] In some embodiments, the 4 IBB intracellular signaling domain(s) can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81 %, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 26.

[00120] In some embodiments, the CD28 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 27.

[00121] In some embodiments, the CD3^ intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 28.

[00122] In some embodiments, the 0X40 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 29.

[00123] In some embodiments, the CD16A intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 30.

[00124] In some embodiments, the yc intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 31 .

[00125] In some embodiments, the 2B4 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 32.

[00126] In some embodiments, the DAP 10 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 33.

[00127] In some embodiments, the DAP 12 intracellular signaling domain can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 34.

[00128] Because distance between T cell and target cell is an important factor affecting tumor recognition and cytotoxicity, a spacer domain can be incorporated between extracellular antigen binding domain and the hinge domain or between the hinge domain and the transmembrane domain. In some embodiments, the spacer comprises a simple alkyl chain, such as (CHTMCHI unit, wherein n is the number of CH2 groups and can vary from 1- 100, preferably 1-50, 1-20, or 1-10. In another example, the spacer can be a peptide of 1-50 amino acids, such as 1-20 amino acids, or 1-10 amino acids.

[00129] In some embodiments, the spacer can include an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 35.

[00130] In another embodiment, the CAR further comprises a signaling peptide.

[00131] In some embodiments, the signaling peptide comprises an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 36 or SEQ ID NO: 37. [00132] In some embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a hinge domain, a transmembrane domain, a CD28 intracellular domain; and an 0X40 intracellular domain.

[00133] In some embodiments, the hinge domain is an IgGl hinge domain, and the transmembrane domain is a CD28 transmembrane domain.

[00134] In other embodiments, the CAR includes, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, an IgGl hinge domain, a CD28 transmembrane domain, the CD28 intracellular domain, the 0X40 intracellular domain, and a CD3-zeta intracellular domain.

[00135] In some embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a CD 16a intracellular signaling domain, a 2B4 intracellular domain, and a CD3^ intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[00136] In other embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a yc intracellular signaling domain, a 2B4 intracellular domain, and a CD3^ intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[00137] In other embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a CD28 intracellular signaling domain, a 2B4 intracellular domain, and a CD3 intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[00138] In other embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a 2B4 intracellular signaling domain, a 4 IBB intracellular domain, and a CD3^ intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[00139] In other embodiments, the CAR can include, from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an TgGl hinge domain, a transmembrane domain, a 2B4 intracellular signaling domain, a DAP10 intracellular domain, a DAP 12 intracellular domain, and a CD3 intracellular domain. The transmembrane domain can be selected from a CD28 transmembrane domain, a CD 16a transmembrane domain, a 2B4 transmembrane domain, or a NKG2D transmembrane domain.

[00140] Other embodiments relate to a nucleic acid comprising a nucleotide sequence encoding a CAR described herein. The nucleic acid encoding the CAR is easily prepared from an amino acid sequence of the specified CAR by any conventional method. A base sequence encoding an amino acid sequence can be obtained from the aforementioned NCBI RefSeq IDs or accession numbers of GenBank for an amino acid sequence of each domain, and the nucleic acid of the present disclosure can be prepared using a standard molecular biological and/or chemical procedure. For example, based on the base sequence, a polynucleotide can be synthesized, and the polynucleotide of the present disclosure can be prepared by combining DNA fragments which are obtained from a cDNA library using a polymerase chain reaction (PCR).

[00141] The CAR encoding nucleotide sequence can be operably linked to a promoter and provided in an expression construct. The vector can be suitable for replication and integration into eukaryotes. Typical vectors contain transcription and translation terminators, initiation sequences, and promoters useful for regulation of the expression of the desired nucleic acid sequence.

[00142] In an embodiment, the vector is a viral vector. Viral vector technology is known in the art and is described, for example, in Sambrook et al., 2012, Molecular Cloning: A Laboratory Manual, volumes 1-4, Cold Spring Harbor Press, NY), and in other virology and molecular biology manuals. In an embodiment, viruses, which are useful as vectors are retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, and lentiviruses. In an embodiment the vector is a lentivirus vector. In general, a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers, (e.g., WO 01/96584; WO 01/29058; and U.S. Pat. No. 6,326,193)

[00143] Vectors derived from viruses, e.g., lentivirus, are suitable tools to achieve longterm gene transfer since they allow long-term, stable integration of a transgene and its propagation in daughter cells. Lentiviral vectors have the added advantage over vectors derived from retroviruses e.g., murine leukemia viruses, in that they can transduce nonproliferating cells, such as hepatocytes. They also have the added advantage of low immunogenicity.

[00144] A retroviral vector may also be, e.g., a gammaretroviral vector. A gammaretroviral vector may include, e.g., a promoter, a packaging signal (\|/), a primer binding site (PBS), one or more (e.g., two) long terminal repeats (LTR), and a transgene of interest, e.g., a gene encoding a CAR. A gammaretroviral vector may lack viral structural gens such as gag, pol, and env. Exemplary gammaretroviral vectors include Murine Leukemia Virus (MLV), Spleen-Focus Forming Virus (SFFV), and Myeloproliferative Sarcoma Virus (MPSV), and vectors derived therefrom.

[00145] In some embodiments, the vector can express two or more genes, where each gene is expressed separately under the control of a different promoter region, e.g., by using bi or tri-cistronic promoters. Expression of two or more genes from the same vector can be achieved by using either a multiple promoter plasmid e.g., bi or tri-cistronic promoters. Examples of multiple promoter containing lentivirus vectors are known in the literature. For example the vector pLENTI-bi-cistronic drives the expression of two genes using the PKG promoter and the mini CMV promoter in opposite directions (Applied Biological Material Inc., Richmond, BC, Canada). Similarly, the tri-cistronic vector pLENTI-tri-cistronic drives expression of three genes. In this configuration one gene can be induced by the mini-CMV promoter while the second and third gene can be induced by the PGK promoter separating the two genes with a T2A peptide cleavage site.

[00146] In another embodiment, bi- or tri-cistronic vectors may also be constructed making use of internal ribosomal entry sites (IRES) such as for example the element from the encephalomyocarditis virus (EMCV) for translation of two or more open reading frames (ORFs). Such vectors are designed to drive transcription of the bi- or tri-cistronic message under control of a strong human promoter regulatory region e.g., CMV or EFl alpha. IRESs are relatively short DNA sequences that can initiate RNA translation in a 5' cap-independent fashion. Whereas the first cistron is translated in a cap-dependent manner driven by a strong mammalian promoter, the subsequent ones utilize intercistronic regions of viral origin such as the internal ribosomal entry site of poliovirus or the cap-independent translation enhancer of encephalomyocarditis virus for enhanced translation.

[00147] Additional promoter elements, e.g., enhancers, can regulate the frequency of transcriptional initiation. Typically, these are located in the region 30-110 bp upstream of the start site, although a number of promoters have been shown to contain functional elements downstream of the start site as well. The spacing between promoter elements frequently is flexible, so that promoter function is preserved when elements are inverted or moved relative to one another. In the thymidine kinase (tk) promoter, the spacing between promoter elements can be increased to 50 bp apart before activity begins to decline. Depending on the promoter, the individual elements can function either cooperatively or independently to activate transcription.

[00148] Other examples of promoters include an SFFV promoter and a cytomegalovirus (CMV) promoter sequence. Other constitutive promoter sequences may also be used, including, but not limited to the simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, an avian leukemia virus promoter, an Epstein-Barr virus immediate early promoter, a Rous sarcoma virus promoter, as well as human gene promoters such as, but not limited to, the actin promoter, the myosin promoter, the elongation factor- la promoter (EFla), the hemoglobin promoter, and the creatine kinase promoter.

[00149] Further, embodiments are not limited to the use of constitutive promoters and can include, for example, inducible promoters. The use of an inducible promoter provides a molecular switch capable of turning on expression of the polynucleotide sequence which it is operatively linked when such expression is desired, or turning off the expression when expression is not desired. Examples of inducible promoters include, but are not limited to a metallothionine promoter, a glucocorticoid promoter, a progesterone promoter, and a tetracycline promoter.

[00150] The vector may also include, e.g., a signal sequence to facilitate secretion, a polyadenylation signal and transcription terminator e.g., from Bovine Growth Hormone (BGH) gene), an element allowing episomal replication and replication in prokaryotes (e.g., SV40 origin and ColEl or others known in the art) and/or elements to allow selection (e.g., puromycin resistant gene, ampicillin resistance gene and/or zeocin marker).

[00151] Sequences encoding various elements of a CAR can be disposed on the same nucleic acid molecule, e.g., the same plasmid or vector, e.g., viral vector, e.g., lentiviral vector. For example, both (i) sequence encoding extracellular antigen binding domain and (ii) sequence encoding an intracellular signaling member, can be present on the same nucleic acid, e.g., vector. Production of the corresponding proteins can be achieved, e.g., by the use of separate promoters, or by the use of a bicistronic transcription product (which can result in the production of two proteins by cleavage of a single translation product or by the translation of two separate protein products).

[00152] In order to assess the expression of a CAR polypeptide or portions thereof, 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 aspects, the selectable marker may be carried on a separate piece of DNA and used in a cotransfection procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers include, for example, antibiotic -resistance genes, such as neo and the like.

[00153] Reporter genes are used for identifying potentially transfected cells and for evaluating the functionality of regulatory sequences. In general, a reporter gene is a gene that is not present in or expressed by the recipient organism or tissue and that encodes a polypeptide whose expression is manifested by some easily detectable property, e.g., enzymatic activity. Expression of the reporter gene is assayed at a suitable time after the DNA has been introduced into the recipient cells. Suitable reporter genes may include genes encoding luciferase, beta-galactosidase, chloramphenicol acetyl transferase, secreted alkaline phosphatase, or the green fluorescent protein gene e.g., Ui-Tei et al., 2000 FEBS Letters 479: 79-82). Suitable expression systems are well known and may be prepared using known techniques or obtained commercially. In general, the construct with the minimal 5' flanking region showing the highest level of expression of reporter gene is identified as the promoter. Such promoter regions may be linked to a reporter gene and used to evaluate agents for the ability to modulate promoter — driven transcription. [00154] Methods of introducing and expressing genes into a cell are known in the art. In the context of an expression vector, the vector can be readily introduced into a host cell, e.g., mammalian, bacterial, yeast, or insect cell by any method in the art. For example, the expression vector can be transferred into a host cell by physical, chemical, or biological means.

[00155] Physical methods for introducing a polynucleotide into a host cell include calcium phosphate precipitation, lipofection, particle bombardment, microinjection, electroporation, and the like. Methods for producing cells comprising vectors and/or exogenous nucleic acids are well-known in the art. See, for example, Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). A preferred method for the introduction of a polynucleotide into a host cell is calcium phosphate transfection.

[00156] Biological methods for introducing a polynucleotide of interest into a host cell include the use of DNA and RNA vectors. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., human cells. Other viral vectors can be derived from lentivirus, poxviruses, herpes simplex virus I, adenoviruses and adeno-associated viruses, and the like. See, for example, U.S. Pat, Nos. 5,350,674 and 5,585,362.

[00157] Chemical means for introducing a polynucleotide into a host cell include colloidal dispersion systems, such as macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system for use as a delivery vehicle in vitro and in vivo is a liposome (e.g., an artificial membrane vesicle). In the case where a non-viral delivery system is utilized, an exemplary delivery vehicle is a liposome. The use of lipid formulations is contemplated for the introduction of the nucleic acids into a host cell in vitro, ex vivo or in vivo). In another aspect, the nucleic acid may be associated with a lipid. The nucleic acid associated with a lipid may be encapsulated in the aqueous interior of a liposome, interspersed within the lipid bilayer of a liposome, attached to a liposome via a linking molecule that is associated with both the liposome and the oligonucleotide, entrapped in a liposome, complexed with a liposome, dispersed in a solution containing a lipid, mixed with a lipid, combined with a lipid, contained as a suspension in a lipid, contained or complexed with a micelle, or otherwise associated with a lipid. Lipid, lipid/DNA or lipid/expression vector associated compositions are not limited to any particular structure in solution. For example, they may be present in a bilayer structure, as micelles, or with a “collapsed” structure. They may also simply be interspersed in a solution, possibly forming aggregates that are not uniform in size or shape. Lipids are fatty substances which may be naturally occurring or synthetic lipids. For example, lipids include the fatty droplets that naturally occur in the cytoplasm as well as the class of compounds which contain long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes.

[00158] Lipids suitable for use can be obtained from commercial sources. For example, dimyristyi phosphatidylcholine (“DMPC”) can be obtained from Sigma, St. Louis, Mo.; dicetyl phosphate (“DCP”) can be obtained from K & K Laboratories (Plainview, N.Y.); cholesterol (“Choi”) can be obtained from Calbiochem-Behring; dimyristyi phosphatidylglycerol (“DMPG”) and other lipids may be obtained from Avanti Polar Lipids, Inc. (Birmingham, Ala.). Stock solutions of lipids in chloroform or chloroform/methanol can be stored at about -20° C. Chloroform is used as the only solvent since it is more readily evaporated than methanol.

[00159] “Liposome” is a generic term encompassing a variety of single and multilamellar lipid vehicles formed by the generation of enclosed lipid bilayers or aggregates. Liposomes can be characterized as having vesicular structures with a phospholipid bilayer membrane and an inner aqueous medium. Multilamellar liposomes have multiple lipid layers separated by aqueous medium. They form spontaneously when phospholipids are suspended in an excess of aqueous solution. The lipid components undergo self-rearrangement before the formation of closed structures and entrap water and dissolved solutes between the lipid bilayers (Ghosh et al., 19 1 Glycobiology 5; 505-10). However, compositions that have different structures in solution than the normal vesicular structure are also encompassed. For example, the lipids may assume a micellar structure or merely exist as nonuniform aggregates of lipid molecules. Also contemplated are lipofectamine — nucleic acid complexes.

[00160] Regardless of the method used to introduce exogenous polynucleotides into a host cell or otherwise expose a cell to the polynucleotide of the present disclosure, in order to confirm the presence of the recombinant DNA sequence in the host cell, a variety of assays may be performed. Such assays include, for example, “molecular biological” assays well known to those of skill in the art, such as Southern and Northern blotting, RT-PCR and PCR; “biochemical” assays, such as detecting the presence or absence of a particular peptide, e.g., by immunological means (ELIS As and Western blots) or by assays described herein to identify agents falling within the scope of the disclosure.

[00161] In another embodiment, the disclosure provides an engineered cell expressing the chimeric antigen receptor polypeptide described above or polynucleotide encoding for the same, and described above.

[00162] An “engineered cell” means any cell of any organism that is modified, transformed, or manipulated by addition or modification of a gene, a DNA or RNA sequence, or protein or polypeptide. Isolated cells, host cells, and genetically engineered cells of the present disclosure include isolated immune cells, such as NK cells and T cells that contain the DNA or RNA sequences encoding a chimeric antigen receptor or chimeric antigen receptor complex and express the chimeric receptor on the cell surface. Isolated host cells and engineered cells may be used, for example, for enhancing an NK cell activity or a T lymphocyte activity, treatment of cancer, and treatment of infectious diseases.

[00163] Any cell capable of expressing and/or capable of integrating the chimeric antigen receptor polypeptide, as disclosed herein, into its membrane may be used.

[00164] In some embodiments, the CAR can be expressed in at least one immune cell. In certain embodiments, the immune cell is a T cell, e.g., a CD8+ T cell e.g., a CD8+ naive T cell, central memory T cell, or effector memory T cell), a CD4+ T cell, a natural killer T cell (NKT cells), a regulatory T cell (Treg), a stem cell memory T cell, a lymphoid progenitor cell a hematopoietic stem cell, a natural killer cell (NK cell) or a dendritic cell. In certain embodiments, the cells are monocytes or granulocytes, e.g., myeloid cells, macrophages, neutrophils, dendritic cells, mast cells, eosinophils, and/or basophils.

[00165] In some embodiments, the immune cells include one or more subsets of T cells or other cell types, such as whole T cell populations, CD4+ cells, CD8+ cells, and subpopulations thereof, such as those defined by function, activation state, maturity, potential for differentiation, expansion, recirculation, localization, and/or persistence capacities, antigen specificity, type of antigen receptor, presence in a particular organ or compartment, marker or cytokine secretion profile, and/or degree of differentiation. Among the sub-types and subpopulations of T cells and/or of CD4+ and/or of CD8+ T cells are naive T (TN) cells, effector T cells (TEFF), memory T cells and sub-types thereof, such as stem cell memory T (TSCM), central memory T (TCM), effector memory T (TEM), or terminally differentiated effector memory T cells, tumor- infiltrating lymphocytes (TIL), immature T cells, mature T cells, helper T cells, cytotoxic T cells, mucosa-associated invariant T (MAIT) cells, naturally occurring and adaptive regulatory T (Treg) cells, helper T cells, such as Thl cells, Th2 cells, Th3 cells, Thl7 cells, Th9 cells, Th22 cells, follicular helper T cells, a/p T cells, and 8/y T cells. In certain embodiments, any number of T cell lines available in the art, may be used. [00166] The engineered cells may be obtained from peripheral blood, cord blood, bone marrow, tumor infiltrating lymphocytes, lymph node tissue, or thymus tissue. The host cells may include placental cells, embryonic stem cells, induced pluripotent stem cells, or hematopoietic stem cells. The cells may be obtained from humans, monkeys, chimpanzees, dogs, cats, mice, rats, and transgenic species thereof. The cells may be obtained from established cell lines.

[00167] The above cells may be obtained by any known means. The cells may be autologous, syngeneic, allogeneic, or xenogeneic to the recipient of the engineered cells. [00168] In certain embodiments, T and NK cells are derived from human peripheral blood mononuclear cells (PBMC), leukapheresis products (PBSC), human embryonic stem cells (hESCs), induced pluripotent stem cells (iPSCs), bone marrow, or umbilical cord blood. [00169] In some embodiments, a plurality of the engineered CAR immune cells can be provided in an immunotherapy composition. The immunotherapy composition can be used in a method of treating cancer in a subject in need thereof. The method includes administering to a subject a therapeutically effective amount of immunotherapy composition comprising the engineered CAR immune cells thereby treating cancer in the subject.

[00170] In particular embodiments, the immunotherapy composition can be used as treatment of virtually all types of cancers and pre-cancers (e.g., Myelodysplastic syndrome), including but not limited to carcinomas, sarcomas, melanomas, lymphomas, and leukemias, and having places of origin including but not limited to colon, prostate, brain, breast, liver, lung, pancreatic, bone, ovarian, skin, pancreatic, blood and others. The methods disclosed herein are contemplated for treatment of both metastatic cancers as well as primary tumor sites.

[00171] The immunotherapy composition as described herein can optionally include a pharmaceutically acceptable carrier. The active ingredients of the pharmaceutical composition at a minimum comprise the immunotherapy, e.g., CAR immune cells as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist essentially of the CAR immune cells as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist of the CAR immune cells as described herein. Pharmaceutically acceptable carriers for cell-based therapeutic formulation include saline and aqueous buffer solutions, Ringer's solution, and serum component, such as serum albumin, HDL and LDL. The terms such as “excipient”, “carrier”, “pharmaceutically acceptable carrier” or the like are used interchangeably herein. [00172] In some embodiments, the immunotherapy composition as described herein can be a parenteral dose form. Since administration of parenteral dosage forms typically bypasses the patient's natural defenses against contaminants, the components apart from the immunotherapies themselves are preferably sterile or capable of being sterilized prior to administration to a patient. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions. Any of these can be added to the immunotherapy preparation prior to administration.

[00173] Suitable vehicles that can be used to provide parenteral dosage forms of immunotherapies as disclosed within are well known to those skilled in the art. Examples include, without limitation: saline solution; glucose solution; aqueous vehicles including but not limited to, sodium chloride injection, Ringer's injection, dextrose Injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and propylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[00174] In some embodiments, the immunotherapy composition described herein can administered as a monotherapy, i.e., another treatment for the condition is not concurrently administered to the subject.

[00175] Modes of administration can include, for example intravenous (i.v.) injection or infusion. The compositions described herein can be administered to a patient transarterially, intratumorally, intranodally, or intramedullary. In some embodiments, the compositions of immunotherapy may be injected directly into a tumor, lymph node, or site of infection. In one embodiment, the compositions described herein are administered into a body cavity or body fluid (e.g., ascites, pleural fluid, peritoneal fluid, or cerebrospinal fluid). [00176] The dosage of the above treatments to be administered to a patient will vary with the precise nature of the condition being treated and the recipient of the treatment. The scaling of dosages for human administration can be performed according to art-accepted practices.

[00177] In some embodiments, a single treatment regimen is required. In others, administration of one or more subsequent doses or treatment regimens can be performed. For example, after treatment biweekly for three months, treatment can be repeated once per month, for six months or a year or longer. In some embodiments, no additional treatments are administered following the initial treatment.

[00178] The dosage of a composition as described herein can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment. With respect to duration and frequency of treatment, it is typical for skilled clinicians to monitor subjects in order to determine when the treatment is providing therapeutic benefit, and to determine whether to administer further cells, discontinue treatment, resume treatment, or make other alterations to the treatment regimen. The dosage should not be so large as to cause adverse side effects, such as cytokine release syndrome. Generally, the dosage will vary with the age, condition, and sex of the patient and can be determined by one of skill in the art. The dosage can also be adjusted by the individual physician in the event of any complication.

[00179] In some embodiments, the immunotherapy composition administered to the subject can include at least about 1 million engineered CAR immune cells, at least about 2 million engineered CAR immune cells, at least about 3 million engineered CAR immune cells, at least about 4 million engineered CAR immune cells, at least about 5 million engineered CAR immune cells, or at least about 10 million engineered CAR immune cells. [00180] In another aspect, a method of treating cancer in a subject in need thereof is provided including administering to a subject a therapeutically effective amount of the immunotherapy composition provided herein, thereby treating cancer in the subject. Optionally, the cancer such as a solid tumor or hematologic malignancy.

[00181] In some embodiments, the solid tumor can be an osteosarcoma, melanoma, mesothelioma, breast cancer, prostate cancer, gastric cancer, head and neck cancer, lung cancer, or brain cancer.

[00182] In other embodiments, the hematologic malignancy can be any hematologic malignancy wherein the cancer cells express or overexpress a receptor of OSM, including but not limited to acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, Acute lymphoblastic leukemia, non-Hodgkin’s lymphoma, Hodgkin’s lymphoma, B cell malignancies, and multiple myeloma.

[00183] Optionally, the method further includes administering to the subject an additional therapeutic in combination with the immunotherapy composition. The additional therapeutic can include other types of therapy for cancer, such as chemotherapy, surgery, radiation, gene therapy, and so forth. Such therapies can be administered simultaneously or sequentially (in any order) with the immunotherapy composition described herein. When coadministered with an additional therapeutic, suitable therapeutically effective dosages for each agent may be lowered due to the additive action or synergy.

[00184] Non-limiting examples of other anti-cancer therapeutic agents useful for combination with the CAR immune cells described herein include, but are not limited to, immune checkpoint inhibitors (e.g., PDL1, PD1, and CTLA4 inhibitors), anti- angiogenic agents e.g., TNP-470, platelet factor 4, thrombospondin- 1 , tissue inhibitors of metalloproteases, prolactin, angiostatin, endostatin, bFGF soluble receptor, transforming growth factor beta, interferon alpha, soluble KDR and FLT-1 receptors, and placental proliferin-related protein); a VEGF antagonist (e.g., anti-VEGF antibodies, VEGF variants, soluble VEGF receptor fragments); chemotherapeutic compounds. Exemplary chemotherapeutic compounds include pyrimidine analogs (e.g., 5 -fluorouracil, floxuridine, capecitabine, gemcitabine and cytarabine); purine analogs (e.g., fludarabine); folate antagonists (e.g., mercaptopurine and thioguanine); antiproliferative or antimitotic agents, for example, vinca alkaloids; microtubule disruptors such as taxane (e.g., paclitaxel, docetaxel), vincristin, vinblastin, nocodazole, epothilones and navelbine, and epidipodophyllotoxins; DNA damaging agents (e.g., actinomycin, amsacrine, an thracy clines, bleomycin, busulfan, camptothecin, carboplatin, chlorambucil, cisplatin, cyclophosphamide, cytoxan, dactinomycin, daunorubicin, doxorubicin, epirubicin, hexamethyhnelamineoxaliplatin, iphosphamide, melphalan, merchlorehtamine, mitomycin, mitoxantrone, nitrosourea, plicamycin, procarbazine, taxol, taxotere, teniposide, triethylenethiophosphoramide and etoposide). [00185] In some embodiments, radiation, or radiation and chemotherapy are used in combination with the cell populations comprising CAR immune cells described herein. Additional useful agents and therapies can be found in Physician's Desk Reference, 59.sup.th edition, (2005), Thomson P D R, Montvale N.J.; Gennaro el al., Eds. Remington’s The Science and Practice of Pharmacy 20.sup.th edition, (2000), Lippincott Williams and Wilkins, Baltimore Md.; Braunwald et al., Eds. Harrison's Principles of Internal Medicine, 15. sup. th edition, (2001), McGraw Hill, N.Y.; Berkow et al., Eds. The Merck Manual of Diagnosis and Therapy, (1992), Merck Research Laboratories, Rahway N.J.

[00186] The following examples are illustrative and are not intended to limit the scope of the invention as claimed.

EXAMPLES

[00187] We designed an oncostatin-M (OSM)-ligand based Chimeric antigen receptor (CAR) T cell. This has the ligand OSM displayed at the surface of a CAR-construct with the purposes of binding to oncostatin m receptor (OSMR) and Leukemic inhibitory factor receptor (LIFR) to target and kill cancer cells and surrounding cells that support these cancer cells in the tumor microenvironment. The following Examples show OSM-CAR-T cells express OSM on the surface and display greater cytotoxicity than untransduced T-cells against a variety of solid cancer cell lines that all express OSMR both in vitro and in vivo. They do not display this cytotoxicity against jeko cells, which lack OSMR, in vitro or in vivo. [00188] Additionally, mutant CARs that bind LIFR with stronger affinity are possible through mutations of the B-C alpha helical regions. These mutant OSM molecules can be displayed as ligands and their sequences are described herein.

[00189] Experimental notes: All experiments were carried out on Nod-Scid-Gamma (NSG) mice between 9-12 weeks of age at the time of injection.

Example 1

[00190] Figs. l(A-H) illustrate graphs showing OSM CARs express OSM and preferentially kill specific cell lines. OSM ligand was added to our third generation lentiviral CAR-T vector (pHR backbone and existing CAR-T internal structure of BAFF-CAR-T cells described in U.S. Pub. No. 2020/0376032 Al, which is incorporated by reference in its entirety) with a GFP reporter that can be measured in the FITC channel on flow cytometry. We utilized an APC-channel flow-cytometry-compatible antibody that recognizes OSM to determine if transduced CAR-T cells were displaying OSM ligand on the surface. Upon confirming OSM expression on the CAR-T cell surface, we looked for cells that expressed OSMR (the binding partner of OSM ligand) on the surface. Importantly, B16F10, a mouse melanoma cell line was utilized as a specificity control showing that our human OSMR flowbased antibody does not bind mouse OSMR. Results were normalized to TgG control antibody, which means a value of 1 would indicate no expression above the control antibody. Jeko cells, a mantle cell lymphoma do not display OSMR. Other solid tumors displayed OSMR at various levels. We next tested cytotoxicity at a 7: 1 ratio (CAR:Tumor cells) for 16 hours against these human cancer cell lines. We found significantly increased cytotoxicity in all cell lines expressing OSMR, but not Jeko cells that did not express OSMR. Further comparison with our lab’s BAFF-CAR-T cells (with identical lentiviral CAR vector except for the external ligand) which target receptors on the surface of Jeko cells showed that OSM- CAR-T cells do not kill Jeko cells, while BAFF CAR-Ts do. We next examined the release of cytokines after 16 hours of co-incubation between OSM-CAR-T cells and cancer cells. We see increases in many cytotoxic and T-cell activity mediating cytokines, but preferential increases in certain mesothelioma (H2052) and osteosarcoma (SAOS2) cell lines. Comparing degranulation via CD 107a, we saw drastic increases in degranulation of our CAR-T cells when exposed to cancer cells expressing OSMR. This was true when comparing CAR-Ts to Untransduced T-cells (UT) that were expanded separately, or when flow gating on the GFP- population of T-cells within the transduced CAR well itself. This importantly controls for T- cells exposed to increased cytokines that the CARs are secreting, and demonstrates that the OSM-CAR-T cells are chiefly responsible for the response to the cancer cells. Note that for all assays the same number of T-cells were added to each well. This means that when 175,000 “OSM-CAR-T-cells” were added to the plate to match the 175,000 Untransduced T- cells, only 40-90% of these were OSM-CAR-T cells depending on transduction efficiency.

Example 2

[00191] Figs. 2(A-C) illustrate an image and graphs showing intratumoral Injection of OSM-CARs reduces solid tumor burden. Two pilot studies were carried out to demonstrate if OSM-CAR-T cells could display similar in vitro cytotoxicity in a solid tumor in vivo environment. AGS-luciferase tagged cells were injected into mice subcutaneously. Their growth was tracked via intraperitoneal D-luciferin injection and imaging on an IVIS Spectrum imager. One mouse had 4,000,000 T-cells (-70% CARs) injected intratumorally, while two mice received PBS intratumorally. We saw complete elimination of the OSM- CAR-T treated tumor upon extraction. Importantly we expanded upon this pilot finding using Detroit 562 cells, a pharyngeal cancer, as this model grew better subcutaneously. Once tumors were palpable, an intratumoral injection of 4,000,000 T-cells (-70% CARs) resulted in statistically significant decreases in tumor volume and mass upon extraction 12-14 days after injection compared to both PBS injection and Untransduced T-cell injection.

Example 3

[00192] Figs. 3(A-E) illustrate a schematic, graph, and plots showing intravenously (IV) injected OSM CAR-T cells display cytotoxicity against subcutaneous SAOS2. To determine if OSM-CAR-T cells display cytotoxicity when injected IV against a subcutaneous solid tumor, we utilized a pediatric osteosarcoma cell line called SAOS2. We injected 4,000,000 T-cells (-50% CARs) IV after subcutaneous SAOS2 tumors (with Matrigel support) were palpable (Day 7). There was a statistically significant weight loss that was correlated to the level of response seen in the mice. As the response lightened, the mass of the mice recovered, and tumors grew. No special interventions (gels, diets, etc.) were taken to combat mouse weight-loss and mice all eventually recovered displaying about a 10 day survival advantage. Overall, there was cytotoxic effects of OSM-CAR-T cells on established SAOS2 subcutaneous tumors.

Example 4

[00193] Figs. 4(A-D) illustrate a schematic, charts, and image showing higher dose of OSM CAR-Ts increases Tumor Killing and Toxicity. We sought to determine if the cytotoxic effects of our OSM-CAR-T cells could be increased by increasing the cell number of T-cells injected and percentage of CAR-T cells injected. We injected 6,000,000 T-cells (-90% CAR-Ts) IV after SAOS2 (with matrigel support) tumors were palpable. A large reduction in tumor burden was seen, including the complete elimination of detectable tumors after only 5 days. The difference between OSM-CAR-T cell’s shrinkage of SAOS2 tumors compared to Untransduced T-cells (UT) and PBS (saline) was highly significant. Untransduced T-cells did not produce statistically significant tumor shrinkage compared to PBS alone. Importantly, these mice displayed even more severe weight loss/toxicity than previous injected mice, that was lethal for some mice. We took serum from these mice to analyze for cytokines and stopped the experiment after only 5 days. Serum displayed a large increase in many human cytokines, only in the OSM-CAR-T mice, that included elevated IFNy. This is a common cytokine associated with CAR-T cytotoxicity against solid tumors.

Example 5

[00194] Figs. 5(A-E) illustrate a schematic and graphs showing intravenously injected OSM CARs display cytotoxicity against subcutaneous 143B. We repeated the schematic of the previous SAOS2 experiment with another osteosarcoma cell line 143B that established a subcutaneous tumor with Matrigel support. 6,000,000 T-cells (70% OSM-CAR-Ts) were injected IV after 7 days when 143B tumors were palpable. Again, we saw a reduction in mass in the OSM-CAR treated mice but supported the mice with Diet-Hydration Gel. Some OSM-CAR treated mice plateaued in weight loss, but others continued to lose mass. All mice displayed highly reduced tumor burden for 21 days after injection of the OSM-CAR-T cells when experiment was stopped to compare tumor volumes as many mice in PBS and UT group were reaching the 2000mm³ experimental cutoff. 2 out of 10 mice in the OSM-CAR-T treated group had to be euthanized due to weight loss/toxicity.

Example 6

[00195] Figs. 6(A-E) illustrate a schematic, images, plot, and graph showing intravenously injected OSM CARs display insignificant cytotoxicity against subcutaneous Jeko cells. To determine specificity of OSM-CAR toxicity, a Jeko-luciferase subcutaneous model was created with 5,000,000 T-cells injected (-50% OSM-CAR-Ts) intravenously. These mice did not display reductions in mass like those seen in SAOS2 and 143B experiments. Importantly, imaging performed with D-luciferin IP injections and analysis on the IVIS Spectrum imager showed that tumor burden was unchanged with OSM-CAR presence. Experiment was stopped at 14 days post injection as tumor burden was approaching 2000mm³ experimental cutoff and no effect of the OSM-CAR T-cells on Jeko cells was seen. SEQUENCES pHR-OSM-CAR 1

AGTGGAGCAAGGCAGGTGGAGCGGCCGCGCCACCATGGCCTTACCAGTGACCGC

CTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCGCCAGGCCGACTAGTGCGGCT

ATAGGCAGCTGCTCGAAAGAGTACCGCGTGCTCCTTGGCCAGCTCCAGAAGCAG

ACAGATCTCATGCAGGACACCAGCAGACTCCTGGACCCCTATATACGTATCCAA

GGCCTGGATGTTCCTAAACTGAGAGAGCACTGCAGGGAGCGCCCCGGGGCCTTC

CCCAGTGAGGAGACCCTGAGGGGGCTGGGCAGGCGGGGCTTCCTGCAGACCCTC

AATGCCACACTGGGCTGCGTCCTGCACAGACTGGCCGACTTAGAGCAGCGCCTC

CCCAAGGCCCAGGATTTGGAGAGGTCTGGGCTGAACATCGAGGACTTGGAGAAG

CTGCAGATGGCGAGGCCGAACATCCTCGGGCTCAGGAACAACATCTACTGCATG

GCCCAGCTGCTGGACAACTCAGACACGGCTGAGCCCACGAAGGCTGGCCGGGGG

GCCTCTCAGCCGCCCACCCCCACCCCTGCCTCGGATGCTTTTCAGCGCAAGCTGG

AGGGCTGCAGGTTCCTGCATGGCTACCATCGCTTCATGCACTCAGTGGGGCGGGT

CTTCAGCAAGTGGGGGGAGAGCCCGAACCGGAGCCGGAGATCTGGAGGCGGCTC

GGATCCCGCCGAGCCCAAATCTCCTGACAAAACTCACACATGCCCACCGTGCCC

AAAAGATCCCAAATTTTGGGTGCTGGTGGTGG (SEQ ID NO: 1) pHR-OSM-CAR 2

AGTGGAGCAAGGCAGGTGGAGCGGCCGCGCCACCATGGCCTTACCAGTGACCGC

CTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCGCCAGGCCGACTAGTGCGGCT

ATAGGCAGCTGCTCGAAAGAGTACCGCGTGCTCCTTGGCCAGCTCCAGAAGCAG

ACAGATCTCATGCAGGACACCAGCAGACTCCTGGACCCCTATATACGTATCCAA

GGCCTGGATGTTCCTAAACTGAGAGAGCACTGCAGGGAGCGCCCCGGGGCCTTC

CCCAGTGAGGAGACCCTGAGGGGGCTGGGCAGGCGGGGCTTCCTGCAGACCCTC

AATGCCACACTGGGCTGCGTCCTGCACAGACTGGCCGACTTAGAGCAGCGCCTC

CCCAAGGCCCAGGATTTGGAGAGGTCTGGGCTGAACATCGAGGACTTGGAGAAG

CTGCAGATGGCGAGGCCGAACATCCTCGGGCTCAGGAACAACATCTACTGCATG

GCCCAGCTGCTGGACAACTCAGACACGGCTGAGCCCACGAAGGCTGGCCGGGGG

GCCTCTCAGCCGCCCACCCCCACCCCTGCCTCGGATGCTTTTCAGCGCAAGCTGG

AGGGCTGCAGGTTCCTGCATGGCTACCATCGCTTCATGCACTCAGTGGGGCGGGT

CTTCAGCAAGTGGGGGGAGAGCCCGAACCGGAGCCGGAGATCTGGAGGCGGCTC

GGATCCCGCCGAGCCCAAATCTCCTGACAAAACTCACACATGCCCACCGTGCCA

AAAGATCCCAAATTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGG (SEQ ID

NO: 2) pHR-OSM-CAR 3

AGTGGAGCAAGGCAGGTGGAGCGGCCGCGCCACCATGGCCTTACCAGTGACCGC

CTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCGCCAGGCCGACTAGTGCGGCT

ATAGGCAGCTGCTCGAAAGAGTACCGCGTGCTCCTTGGCCAGCTCCAGAAGCAG

ACAGATCTCATGCAGGACACCAGCAGACTCCTGGACCCCTATATACGTATCCAA

GGCCTGGATGTTCCTAAACTGAGAGAGCACTGCAGGGAGCGCCCCGGGGCCTTC

CCCAGTGAGGAGACCCTGAGGGGGCTGGGCAGGCGGGGCTTCCTGCAGACCCTC

AATGCCACACTGGGCTGCGTCCTGCACAGACTGGCCGACTTAGAGCAGCGCCTC

CCCAAGGCCCAGGATTTGGAGAGGTCTGGGCTGAACATCGAGGACTTGGAGAAG CTGCAGATGGCGAGGCCGAACATCCTCGGGCTCAGGAACAACATCTACTGCATG GCCCAGCTGCTGGACAACTCAGACACGGCTGAGCCCACGAAGGCTGGCCGGGGG GCCTCTCAGCCGCCCACCCCCACCCCTGCCTCGGATGCTTTTCAGCGCAAGCTGG AGGGCTGCAGGTTCCTGCATGGCTACCATCGCTTCATGCACTCAGTGGGGCGGGT CTTCAGCAAGTGGGGGGAGAGCCCGAACCGGAGCCGGAGATCTGGAGGCGGCTC GGATCCCGCCGAGCCCAAATCTCCTGACAAAACTCACACATGCCCACCGTGCCA AAAGATCCCAAATTTTGGGTGCTGGTGGTGGTTTGGTGGAGTCCTGG (SEQ ID NO: 3) pLVX-EcoRI-SpeI-NdeI-P2A-XmaI-SmaI-XbaI-BamHI-T2A-NheI-NsiI-HpaI-EGFP-SalI- Mlul

OSM-WITHOUT-PROPEPTIDE tggaagggctaattcactcccaaagaagacaagatatccttgatctgtggatctaccacacacaaggctacttccctgattagcagaact acacaccagggccaggggtcagatatccactgacctttggatggtgctacaagctagtaccagttgagccagataaggtagaagagg ccaataaaggagagaacaccagcttgttacaccctgtgagcctgcatgggatggatgacccggagagagaagtgttagagtggaggt ttgacagccgcctagcatttcatcacgtggcccgagagctgcatccggagtacttcaagaactgctgatatcgagcttgctacaaggga ctttccgctggggactttccagggaggcgtggcctgggcgggactggggagtggcgagccctcagatcctgcatataagcagctgct ttttgcctgtactgggtctctctggttagaccagatctgagcctgggagctctctggctaactagggaacccactgcttaagcctcaataa agcttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggtaactagagatccctcagacccttttagtcagtgtggaa aatctctagcagtggcgcccgaacagggacttgaaagcgaaagggaaaccagaggagctctctcgacgcaggactcggcttgctga agcgcgcacggcaagaggcgaggggcggcgactggtgagtacgccaaaaattttgactagcggaggctagaaggagagagatgg gtgcgagagcgtcagtattaagcgggggagaattagatcgcgatgggaaaaaattcggttaaggccagggggaaagaaaaaatata aattaaaacatatagtatgggcaagcagggagctagaacgattcgcagttaatcctggcctgttagaaacatcagaaggctgtagacaa atactgggacagctacaaccatcccttcagacaggatcagaagaacttagatcattatataatacagtagcaaccctctattgtgtgcatc aaaggatagagataaaagacaccaaggaagctttagacaagatagaggaagagcaaaacaaaagtaagaccaccgcacagcaag cggccggccgctgatcttcagacctggaggaggagatatgagggacaattggagaagtgaattatataaatataaagtagtaaaaatt gaaccattaggagtagcacccaccaaggcaaagagaagagtggtgcagagagaaaaaagagcagtgggaataggagctttgttcct tgggttcttgggagcagcaggaagcactatgggcgcagcgtcaatgacgctgacggtacaggccagacaattattgtctggtatagtg cagcagcagaacaatttgctgagggctattgaggcgcaacagcatctgttgcaactcacagtctggggcatcaagcagctccaggca agaatcctggctgtggaaagatacctaaaggatcaacagctcctggggatttggggttgctctggaaaactcatttgcaccactgctgtg ccttggaatgctagttggagtaataaatctctggaacagatttggaatcacacgacctggatggagtgggacagagaaattaacaattac acaagcttaatacactccttaattgaagaatcgcaaaaccagcaagaaaagaatgaacaagaattattggaattagataaatgggcaag tttgtggaattggtttaacataacaaattggctgtggtatataaaattattcataatgatagtaggaggcttggtaggtttaagaatagtttttg ctgtactttctatagtgaatagagttaggcagggatattcaccattatcgtttcagacccacctcccaaccccgaggggacccgacaggc ccgaaggaatagaagaagaaggtggagagagagacagagacagatccattcgattagtgaacggatctcgacggtatcgcctttaaa agaaaaggggggattggggggtacagtgcaggggaaagaatagtagacataatagcaacagacatacaaactaaagaattacaaaa acaaattacaaaaattcaaaattttcgggtttattacagggacagcagagatccagtttatcgatgagtaattcatacaaaaggactcgcc cctgccttggggaatcccagggaccgtcgttaaactcccactaacgtagaacccagagatcgctgcgttcccgccccctcacccgcc cgctctcgtcatcactgaggtggagaagagcatgcgtgaggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtccc cgagaagttggggggaggggtcggcaattgaaccggtgcctagagaaggtggcgcggggtaaactgggaaagtgatgtcgtgtac tggctccgcctttttcccgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaacgttctttttcgcaacgggtttgccgcc agaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacgggttatggcccttgcgtgccttgaattacttccacgccc ctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaaggagccccttcgc ctcgtgcttgagttgaggcctggcttgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctgtctcgctgctttcgat aagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgggccaagatctgcacact ggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcctgcgagcgc ggccaccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggcctcgcgccgccgtgtatcgccccgccc tgggcggcaaggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaaat ggaggacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatg tgactccacggagtaccgggcgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggt tttatgcgatggagtttccccacactgagtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgcccttt ttgagtttggatcttggttcattctcaagcctcagacagtggttcaaagtttttttcttccatttcaggtgtcgtgaggatctatttccggtgaat tcGCCACCATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTC CACGCCGCCAGGCCGACTAGTGCGGCTATAGGCAGCTGCTCGAAAGAGTACCGC GTGCTCCTTGGCCAGCTCCAGAAGCAGACAGATCTCATGCAGGACACCAGCAGA CTCCTGGACCCCTATATACGTATCCAAGGCCTGGATGTTCCTAAACTGAGAGAGC ACTGCAGGGAGCGCCCCGGGGCCTTCCCCAGTGAGGAGACCCTGAGGGGGCTGG GCAGGCGGGGCTTCCTGCAGACCCTCAATGCCACACTGGGCTGCGTCCTGCACA GACTGGCCGACTTAGAGCAGCGCCTCCCCAAGGCCCAGGATTTGGAGAGGTCTG GGCTGAACATCGAGGACTTGGAGAAGCTGCAGATGGCGAGGCCGAACATCCTCG GGCTCAGGAACAACATCTACTGCATGGCCCAGCTGCTGGACAACTCAGACACGG CTGAGCCCACGAAGGCTGGCCGGGGGGCCTCTCAGCCGCCCACCCCCACCCCTG CCTCGGATGCTTTTCAGCGCAAGCTGGAGGGCTGCAGGTTCCTGCATGGCTACCA TCGCTTCATGCACTCAGTGGGGCGGGTCTTCAGCAAGTGGGGGGAGAGCCCGAA CCGGAGCCGGAGATCTGGAGGCGGCTCGGATCCCGCCGAGCCCAAATCTCCTGA CAAAACTCACACATGCCCACCGTGCCCAAAAGATCCCAAATTTTGGGTGCTGGTG GTGGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTAT TTTCTGGGTGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACAT GACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCA CGCGACTTCGCAGCCTATCGCTCCAGGGACCAGAGGCTGCCCCCCGATGCCCAC AAGCCCCCTGGGGGAGGCAGTTTCCGGACCCCCATCCAAGAGGAGCAGGCCGAC GCCCACTCCACCCTGGCCAAGATCAGAGTGAAGTTCAGCAGGAGCGCAGACGCC CCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGA AGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGG GGAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAA AGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGA GGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACA CCTACGACGCCCTTCACATGCAGGCCCTGCCTCCTCGCcatatgggatctggagcaacaaacttct cactactcaaacaagcaggtgacgtggaggagaatcccgggccttctagaggatccggctccggcgagggcaggggaagtctact aacatgcggggacgtggaggaaaatcccggcccagctagcatgcatgttaacatggtgagcaagggcgaggagctgttcaccggg gtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgatgccacct acggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacctacggc gtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccaggagcgc accatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcatcgagct gaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctatatcatgg ccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccgaccacta ccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagtccgccctgagcaaagac cccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgtacaagtg agtcgacacgcgtctggaacaatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatg tggatacgctgctttaatgcctttgtatcatgctattgcttcccgtatggctttcattttctcctccttgtataaatcctggttgctgtctctttatga ggagttgtggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccaccacct gtcagctcctttccgggactttcgctttccccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctggacagggg ctcggctgttgggcactgacaattccgtggtgttgtcggggaagctgacgtcctttccatggctgctcgcctgtgttgccacctggattct gcgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctctgcggcctcttc cgcgtcttcgccttcgccctcagacgagtcggatctccctttgggccgcctccccgcctggaattaattctgcagtcgagacctagaaa aacatggagcaatcacaagtagcaatacagcagctaccaatgctgattgtgcctggctagaagcacaagaggaggaggaggtgggt tttccagtcacacctcaggtacctttaagaccaatgacttacaaggcagctgtagatcttagccactttttaaaagaaaagaggggactg gaagggctaattcactcccaacgaagacaagatatccttgatctgtggatctaccacacacaaggctacttccctgattagcagaactac acaccagggccaggggtcagatatccactgacctttggatggtgctacaagctagtaccagttgagccagataaggtagaagaggcc aataaaggagagaacaccagcttgttacaccctgtgagcctgcatgggatggatgacccggagagagaagtgttagagtggaggttt gacagccgcctagcatttcatcacgtggcccgagagctgcatccggagtacttcaagaactgctgatatcgagcttgctacaagggac tttccgctggggactttccagggaggcgtggcctgggcgggactggggagtggcgagccctcagatcctgcatataagcagctgcttt ttgcctgtactgggtctctctggttagaccagatctgagcctgggagctctctggctaactagggaacccactgcttaagcctcaataaa gcttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggtaactagagatccctcagacccttttagtcagtgtggaaa atctctagcagtagtagttcatgtcatcttattattcagtatttataacttgcaaagaaatgaatatcagagagtgagaggccttgacattgct agcgtttaccgtcgacctctagctagagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacaca acatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgct ttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccg cttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccaca gaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcg tttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagat accaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcggg aagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccc cgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccaclggcagcagcc actggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaa cagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggta gcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgct cagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagtttt aaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgtt catccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcga gacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatcc gcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacagg catcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgc aaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcata attctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccg agttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcgggg cgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcacc agcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatact cttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggt tccgcgcacatttccccgaaaagtgccacctgacgtcgacggatcgggagatcaacttgtttattgcagcttataatggttacaaataaa gcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctgga tcaactggataactcaagctaaccaaaatcatcccaaacttcccaccccataccctattaccactgccaattacctgtggtttcatttactct aaacctgtgattcctctgaattattttcattttaaagaaattgtatttgttaaatatgtactacaaacttagtagt (SEQ ID NO: 4)

OSM-WITH-PROPEPTIDE

ATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCG CCAGGCCGACTAGTGCGGCTATAGGCAGCTGCTCGAAAGAGTACCGCGTGCTCC TTGGCCAGCTCCAGAAGCAGACAGATCTCATGCAGGACACCAGCAGACTCCTGG ACCCCTATATACGTATCCAAGGCCTGGATGTTCCTAAACTGAGAGAGCACTGCAG GGAGCGCCCCGGGGCCTTCCCCAGTGAGGAGACCCTGAGGGGGCTGGGCAGGCG GGGCTTCCTGCAGACCCTCAATGCCACACTGGGCTGCGTCCTGCACAGACTGGCC

GACTTAGAGCAGCGCCTCCCCAAGGCCCAGGATTTGGAGAGGTCTGGGCTGAAC

ATCGAGGACTTGGAGAAGCTGCAGATGGCGAGGCCGAACATCCTCGGGCTCAGG

AACAACATCTACTGCATGGCCCAGCTGCTGGACAACTCAGACACGGCTGAGCCC

ACGAAGGCTGGCCGGGGGGCCTCTCAGCCGCCCACCCCCACCCCTGCCTCGGAT

GCTTTTCAGCGCAAGCTGGAGGGCTGCAGGTTCCTGCATGGCTACCATCGCTTCA

TGCACTCAGTGGGGCGGGTCTTCAGCAAGTGGGGGGAGAGCCCGAACCGGAGCC

GGAGACACAGCCCCCACCAGGCCCTGAGGAAGGGGGTGCGCAGGACCAGACCC

TCCAGGAAAGGCAAGAGACTCATGACCAGGGGACAGCTGCCCCGGTCTGGAGGC

GGCTCGGATCCCGCCGAGCCCAAATCTCCTGACAAAACTCACACATGCCCACCGT

GCCCAAAAGATCCCAAATTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGCTTG

CTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGG

AGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCC

ACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCT

CCAGGGACCAGAGGCTGCCCCCCGATGCCCACAAGCCCCCTGGGGGAGGCAGTT

TCCGGACCCCCATCCAAGAGGAGCAGGCCGACGCCCACTCCACCCTGGCCAAGA

TCAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGA

ACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGG

ACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAAC

CCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTAC

AGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCT

TTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCA

GGCCCTGCCTCCTCGC (SEQ ID NO: 5)

OSM-WITHOUT-PROPEPTIDE-WITH-IL15

ATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCG

CCAGGCCGACTAGTGCGGCTATAGGCAGCTGCTCGAAAGAGTACCGCGTGCTCC

TTGGCCAGCTCCAGAAGCAGACAGATCTCATGCAGGACACCAGCAGACTCCTGG

ACCCCTATATACGTATCCAAGGCCTGGATGTTCCTAAACTGAGAGAGCACTGCAG

GGAGCGCCCCGGGGCCTTCCCCAGTGAGGAGACCCTGAGGGGGCTGGGCAGGCG

GGGCTTCCTGCAGACCCTCAATGCCACACTGGGCTGCGTCCTGCACAGACTGGCC

GACTTAGAGCAGCGCCTCCCCAAGGCCCAGGATTTGGAGAGGTCTGGGCTGAAC

ATCGAGGACTTGGAGAAGCTGCAGATGGCGAGGCCGAACATCCTCGGGCTCAGG

AACAACATCTACTGCATGGCCCAGCTGCTGGACAACTCAGACACGGCTGAGCCC

ACGAAGGCTGGCCGGGGGGCCTCTCAGCCGCCCACCCCCACCCCTGCCTCGGAT

GCTTTTCAGCGCAAGCTGGAGGGCTGCAGGTTCCTGCATGGCTACCATCGCTTCA

TGCACTCAGTGGGGCGGGTCTTCAGCAAGTGGGGGGAGAGCCCGAACCGGAGCC

GGAGATCTGGAGGCGGCTCGGATCCCGCCGAGCCCAAATCTCCTGACAAAACTC

ACACATGCCCACCGTGCCCAAAAGATCCCAAATTTTGGGTGCTGGTGGTGGTTGG

TGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGG

TGAGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCC

GCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTT

CGCAGCCTATCGCTCCAGGGACCAGAGGCTGCCCCCCGATGCCCACAAGCCCCC

TGGGGGAGGCAGTTTCCGGACCCCCATCCAAGAGGAGCAGGCCGACGCCCACTC

CACCCTGGCCAAGATCAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTA

CCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGA

GTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCC GAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGA TGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAG GGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGAC GCCCTTCACATGCAGGCCCTGCCTCCTCGCcatatgggatctggagcaacaaacttctcactactcaaaca agcaggtgacgtggaggagaatcccgggccttctagaatgagaatttcgaaaccacatttgagaagtatttccatccagtgctacttgtg tttacttctaaacagtcattttctaactgaagctggcattcatgtcttcattttgggctgtttcagtgcagggcttcctaaaacagaagccaac tgggtgaatgtaataagtgatttgaaaaaaattgaagatcttattcaatctatgcatattgatgctactttatatacggaaagtgatgttcacc ccagttgcaaagtaacagcaatgaagtgctttctcttggagttacaagttatttcacttgagtccggagatgcaagtattcatgatacagta gaaaatctgatcatcctagcaaacaacagtttgtcttctaatgggaatgtaacagaatctggatgcaaagaatgtgaggaactggagga aaaaaatattaaagaatttttgcagagttttgtacatattgtccaaatgttcatcaacacttctggatccggctccggcgagggcagggga agtctactaacatgcggggacgtggaggaaaatcccggcccagctagcatgcatgttaacatggtgagcaagggcgaggagctgttc accggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaagttcagcgtgtccggcgagggcgagggcgat gccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcccgtgccctggcccaccctcgtgaccaccctgacc tacggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcacgacttcttcaagtccgccatgcccgaaggctacgtccag gagcgcaccatcttcttcaaggacgacggcaactacaagacccgcgccgaggtgaagttcgagggcgacaccctggtgaaccgcat cgagctgaagggcatcgacttcaaggaggacggcaacatcctggggcacaagctggagtacaactacaacagccacaacgtctata tcatggccgacaagcagaagaacggcatcaaggtgaacttcaagatccgccacaacatcgaggacggcagcgtgcagctcgccga ccactaccagcagaacacccccatcggcgacggccccgtgctgctgcccgacaaccactacctgagcacccagtccgccctgagc aaagaccccaacgagaagcgcgatcacatggtcctgctggagttcgtgaccgccgccgggatcactctcggcatggacgagctgta caagtgagtcgacacgcgtctggaacaatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactatgttgctcctttta cgctatgtggatacgctgctUaatgccUtgtatcatgctattgcttcccgtatggctttcattttctcctccttgtataaatcctggttgctgtct ctttatgaggagttgtggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgcc accacctgtcagctcctttccgggactttcgctttccccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctgg acaggggctcggctgttgggcactgacaattccgtggtgttgtcggggaagctgacgtcctttccatggctgctcgcctgtgttgccac ctggattctgcgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctctgc ggcctcttccgcgtcttcgccttcgccctcagacgagtcggatctccctttgggccgcctccccgcctggaattaattctgcagtcgaga cctagaaaaacatggagcaatcacaagtagcaatacagcagctaccaatgctgattgtgcctggctagaagcacaagaggaggagg aggtgggttttccagtcacacctcaggtacctttaagaccaatgacttacaaggcagctgtagatcttagccactttttaaaagaaaagag gggactggaagggctaattcactcccaacgaagacaagatatccttgatctgtggatctaccacacacaaggctacttccctgattagc agaactacacaccagggccaggggtcagatatccactgacctttggatggtgctacaagctagtaccagttgagccagataaggtaga agaggccaataaaggagagaacaccagcttgttacaccctgtgagcctgcatgggatggatgacccggagagagaagtgttagagt ggaggtttgacagccgcctagcatttcatcacgtggcccgagagctgcatccggagtacttcaagaactgctgatatcgagcttgctac aagggactttccgctggggactttccagggaggcgtggcctgggcgggactggggagtggcgagccctcagatcctgcatataagc agctgctttttgcctgtactgggtctctctggttagaccagatctgagcctgggagctctctggctaactagggaacccactgcttaagcc tcaataaagcttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggtaactagagatccctcagacccttttagtcagt gtggaaaatctctagcagtagtagttcatgtcatcttattattcagtatttataacttgcaaagaaatgaatatcagagagtgagaggccttg acattgctagcgtttaccgtcgacctctagctagagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaatt ccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcact gcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcg ctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggtt atccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgtt gctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggact ataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctc ccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcac gaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggc agcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacact agaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccac cgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggt ctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaa tgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgt ctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatga taccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgc aactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccat tgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatccccc atgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcag cactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgc ggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgt tcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttt tactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaat actcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaaca aataggggttccgcgcacatttccccgaaaagtgccacctgacgtcgacggatcgggagatcaacttgtttattgcagcttataatggtt acaaataaagcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatcttat catgtctggatcaactggataactcaagctaaccaaaatcatcccaaacttcccaccccataccctattaccactgccaattacctgtggtt tcatttactctaaacctgtgattcctctgaattattttcattttaaagaaattgtatttgttaaatatgtactacaaacttagtagt (SEQ ID NO: 6)

Uniprot ligand OSM sequence:

AAIGSCSKEYRVLLGQLQKQTDLMQDTSRLLDPYIRIQGLDVPKLREHCRERPGAFPS EETLRGLGRRGFLQTLNATLGCVLHRLADLEQRLPKAQDLERSGLNIEDLEKLQMAR PNILGLRNNIYCMAQLLDNSDTAEPTKAGRGASQPPTPTPASDAFQRKLEGCRFLHG YHRFMHSVGRVFSKWGESPNRSRR (SEQ ID NO: 7)

Pro-Peptide Sequence Uniprot

HSPHQALRKGVRRTRPSRKGKRLMTRGQLPR (SEQ ID NO: 8)

OSM full sequence

Atgggggtactgctcacacagaggacgctgctcagtctggtccttgcactcctgtttccaagcatggcgagcatggcggctataggca gctgctcgaaagagtaccgcgtgctccttggccagctccagaagcagacagatctcatgcaggacaccagcagactcctggacccct atatacgtatccaaggcctggatgttcctaaactgagagagcactgcagggagcgccccggggccttccccagtgaggagaccctga gggggctgggcaggcggggcttcctgcagaccctcaatgccacactgggctgcgtcctgcacagactggccgacttagagcagcg cctccccaaggcccaggatttggagaggtctgggctgaacatcgaggacttggagaagctgcagatggcgaggccgaacatcctcg ggctcaggaacaacatctactgcatggcccagctgctggacaactcagacacggctgagcccacgaaggctggccggggggcctc tcagccgcccacccccacccctgcctcggatgcttttcagcgcaagctggagggctgcaggttcctgcatggctaccatcgcttcatgc actcagtggggcgggtcttcagcaagtggggggagagcccgaaccggagccggagacacagcccccaccaggccctgaggaag ggggtgcgcaggaccagaccctccaggaaaggcaagagactcatgaccaggggacagctgccccggtag (SEQ ID NO: 9)

OSM, no PRO-PEPTIDE or SIGNAL PEPTIDE:

Gcggctataggcagctgctcgaaagagtaccgcgtgctccttggccagctccagaagcagacagatctcatgcaggacaccagca gactcctggacccctatatacgtatccaaggcctggatgttcctaaactgagagagcactgcagggagcgccccggggccttcccca gtgaggagaccctgagggggctgggcaggcggggcttcctgcagaccctcaatgccacactgggctgcgtcctgcacagactggc cgacttagagcagcgcctccccaaggcccaggatttggagaggtctgggctgaacatcgaggacttggagaagctgcagatggcga ggccgaacatcctcgggctcaggaacaacatctactgcatggcccagctgctggacaactcagacacggctgagcccacgaaggct ggccggggggcctctcagccgcccacccccacccctgcctcggatgcttttcagcgcaagctggagggctgcaggttcctgcatgg ctaccatcgcttcatgcactcagtggggcgggtcttcagcaagtggggggagagcccgaaccggagccggaga (SEQ ID NO: 10)

OSM WITH PRO-PEPTIDE

Gcggctataggcagctgctcgaaagagtaccgcgtgctccttggccagctccagaagcagacagatctcatgcaggacaccagca gactcctggacccctatatacgtatccaaggcctggatgttcctaaactgagagagcactgcagggagcgccccggggccttcccca gtgaggagaccctgagggggctgggcaggcggggcttcctgcagaccctcaatgccacactgggctgcgtcctgcacagactggc cgacttagagcagcgcctccccaaggcccaggatttggagaggtctgggctgaacatcgaggacttggagaagctgcagatggcga ggccgaacatcctcgggctcaggaacaacatctactgcatggcccagctgctggacaactcagacacggctgagcccacgaaggct ggccggggggcctctcagccgcccacccccacccctgcctcggatgcttttcagcgcaagctggagggctgcaggttcctgcatgg ctaccatcgcttcatgcactcagtggggcgggtcttcagcaagtggggggagagcccgaaccggagccggagacacagcccccac caggccctgaggaagggggtgcgcaggaccagaccctccaggaaaggcaagagactcatgaccaggggacagctgccccgg (SEQ ID NO: 11)

Additional mutants for targeting LIFR with higher affinity:

Gcggctataggcagctgctcgaaagagtaccgcgtgctccttggccagctccagaagcagacagatctcatgcaggacaccagca gactcctggacccctatatacgtatccaaggcctggatgttcctaaactgagagagcactgcagggagcgccccggggccttcccca gtgaggagaccctgagggggctgggcaggcggggcttcctgcagaccctcaatgccacactgggctgcgtcctgcacagactggc cgacttagagcagcgcctccccaaggcccaggatttggagaggtctgggctgaacatcgaggacttggagaagctgcagatggcga ggccgaacatcctcgggctcaggaacaacatctactgcatggcccagctgctggacaactcagacacggctgagcccacgaaggct ggccggggggcctctcagccgcccacccccacccctgcctcggatgcttttcagcgcaagctggagggctgcaggttcctgcatgg ctaccatcgcttcatgcactcagtggggcgggtcttcagcaagtggggggagagcccgaaccggagccggaga (SEQ ID NO: 12)

Mutant 1

AAIGSCSKEYRVLLGQLQKQTDLMQDTSRLLDPY1R1QGLDVPKLREHCRERPGAFPS EETLRGLGRRGFLQTLNATLGCVLHRLADLEQRLGAPQDLERSGLNIEDLEKLQMA RPNILGLRNNIYCMAQLLDNSDTAEPTKAGRGASQPPTPTPASDAFQRKLEGCRFLH GYHRFMHSVGRVFSKWGESPNRSRR (SEQ ID NO: 13)

MUTANT 1 DNA Sequence

Gcggctataggcagctgctcgaaagagtaccgcgtgctccttggccagctccagaagcagacagatctcatgcaggacaccagca gactcctggacccctatatacgtatccaaggcctggatgttcctaaactgagagagcactgcagggagcgccccggggccttcccca gtgaggagaccctgagggggctgggcaggcggggcttcctgcagaccctcaatgccacactgggctgcgtcctgcacagactggc cgacttagagcagcgcctcGGCGCGCCCtctgggctgaacatcgaggacttggagaagctgcagatggcgaggccgaacat cctcgggctcaggaacaacatctactgcatggcccagctgctggacaactcagacacggctgagcccacgaaggctggccggggg gcctctcagccgcccacccccacccctgcctcggatgcttttcagcgcaagctggagggctgcaggttcctgcatggctaccatcgctt catgcactcagtggggcgggtcttcagcaagtggggggagagcccgaaccggagccggaga (SEQ ID NO: 14)

Mutant 2

AAIGSCSKEYRVLLGQLQKQTDLMQDTSRLLDPYIRIQGLDVPKLREHCRERPGAFPS EETLRGLGRRGFLQTLNATLGCVLHRLADLEQRLGAPSGLNIEDLEKLQMARPNILG LRNNIYCMAQLLDNSDTAEPTKAGRGASQPPTPTPASDAFQRKLEGCRFLHGYHRF MHSVGRVFSKWGESPNRSRR (SEQ ID NO: 15) Mutant 3

AAIGSCSKEYRVLLGQLQKQTDLMQDTSRLLDPYIRIQGLDVPKLREHCRERPGAFPS EETLRGLGRRGFLQTLNATLGCVLHRLADLEQRLGGGQDLERSGLNIEDLEKLQMA RPNILGLRNNIYCMAQLLDNSDTAEPTKAGRGASQPPTPTPASDAFQRKLEGCRFLH GYHRFMHSVGRVFSKWGESPNRSRR (SEQ ID NO: 16)

MUTANT 3 DNA Sequence

Gcggctataggcagctgctcgaaagagtaccgcgtgctccttggccagctccagaagcagacagatctcatgcaggacaccagca gactcctggacccctatatacgtatccaaggcctggatgttcctaaactgagagagcactgcagggagcgccccggggccttcccca gtgaggagaccctgagggggctgggcaggcggggcttcctgcagaccctcaatgccacactgggctgcgtcctgcacagactggc cgacttagagcagcgcctcGGCGGGGGCaacatcgaggacttggagaagctgcagatggcgaggccgaacatcctcgggc tcaggaacaacatctactgcatggcccagctgctggacaactcagacacggctgagcccacgaaggctggccggggggcctctca gccgcccacccccacccctgcctcggatgcttttcagcgcaagctggagggctgcaggttcctgcatggctaccatcgcttcatgcact cagtggggcgggtcttcagcaagtggggggagagcccgaaccggagccggaga (SEQ ID NO: 17)

Mutant 4

AAIGSCSKEYRVLLGQLQKQTDLMQDTSRLLDPYIR1QGLDVPKLREHCRERPGAFPS

EETLRGLGRRGFLQTLNATLGCVLHRLADLEQRLGGGNIEDLEKLQMARPNILGLRN

NIYCMAQLLDNSDTAEPTKAGRGASQPPTPTPASDAFQRKLEGCRFLHGYHRFMHS

VGRVFSKWGESPNRSRR (SEQ ID NO: 18)

CD8a hinge

TTTPAPRPPT PAPTIASQPL SLRPEACRPA AGGAVHTRGL DFACD (SEQ ID NO: 19)

IgGl Hinge

AEPKSPDKTHTCPPCPKDPK (SEQ ID NO: 20)

CD28 Transmembrane

FWVLVVVGGVLACYSLLVTVAFIIFWV (SEQ ID NO: 21)

CD8cc Transmembrane

IYIWAPLAGT CGVLLLSLVI TLYC (SEQ ID NO: 22)

CD16A Transmembrane

VSFCLVMVLLFAVDTGLYFSV (SEQ ID NO: 23)

2B4 Transmembrane

FLVIIVILSALFLGTLACFCV (SEQ ID NO: 24)

NKG2D Transmembrane

PFFFCCFIAVAMGIRFIIMVA (SEQ ID NO: 25) Intracellular 41 BB

KRGRKKLLYI FKQPFMRPVQ TTQEEDGCSC RFPEEEEGGC EL (SEQ ID NO: 26) CD28 intracellular domain

RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS (SEQ ID NO: 27)

Intracellular

RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQ EGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQAL PPR (SEQ ID NO: 28)

Intracellular 0X40

RDQRLPPDAH KPPGGGSFRT PIQEEQADAH STLAKI (SEQ ID NO: 29)

Intracellular CD16A

KTNIRSSTRDWKDHKFKWRKDPQDK (SEQ ID NO: 30)

Intracellular common gamma chain (yc)

ERTMPRIPTLKNLEDLVTEYHGNFSAWSGVSKGLAESLQPDYSERLCLVSEIPPKGGA LGEGPGASPCNQHSPYWAPPCYTLKPET (SEQ ID NO: 31)

Intracellular 2B4

WRRKRKEKQSETSPKEFLTIYEDVKDLKTRRNHEQEQTFPGGGSTIYSMIQSQSSAPT SQEPAYTLYSLIQPSRKSGSRKRNHSPSFNSTIYEVIGKSQPKAQNPARLSRKELENFD VYS (SEQ ID NO: 32)

DAP 10 intracellular domain

LCARPRRSPAQEDGKVY1NMPGRG (SEQ ID NO: 33)

DAP 12 intracellular domain

YFLGRLVPRGRGAAEAATRKQRITETESPYQELQGQRSDVYSDLNTQRPYYK (SEQ ID NO: 34)

Spacer

SGGGSDP (SEQ ID NO: 35)

SIGNALING PEPTIDE

METDTLLLWV LLLWVPGSTG (SEQ ID NO: 36) MALPVTALLL PLALLLHAAR P (SEQ ID NO: 37)

[00196] From the above description of the invention, those skilled in the art will perceive improvements, changes and modifications. Such improvements, changes and modifications within the skill of the art are intended to be covered by the appended claims. All references, publications, and patents cited in the present application are herein incorporated by reference in their entirety.

Claims

Having described the invention, the following is claimed:

1. A chimeric antigen receptor (CAR) comprising: an extracellular antigen binding domain, wherein the extracellular antigen binding domain comprises a polypeptide that binds to an oncostatin-M receptor (OSMR), leukemic inhibitory factor receptor (LIFR), a heterodimer between transmembrane glycoprotein 130 (gpl 30) receptor and OSMR, and/or heterodimer between gpl 30 receptor and LIFR.

2. The CAR of claim 1 , wherein the OSMR and/or LIFR is expressed by cancer cells or other cells in a tumor microenvironment.

3. The CAR of claim 1 or 2, wherein the polypeptide is an oncostatin-M (OSM) ligand that binds to OSMR and/or LIFR and/or heterodimers of OSMR/gpl30 receptor and/or LIFR/gpl30 receptor.

4. The CAR of claim 3, wherein the OSM ligand includes a polypeptide including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 7, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 16, or SEQ ID NO: 18.

5. The CAR of claim 1 or 2, wherein the polypeptide is a single chain variable fragment (ScFv) that binds to OSMR and/or LIFR.

6. The CAR of any of claims 1 to 5, further comprising a hinge domain.

7. The CAR of claim 6, wherein the hinge domain is selected from a hinge domain of CD8a including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 19 or a hinge domain of IgGl including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 20.

8. The CAR of any of claims 1 to 7, further comprising one or more transmembrane domains.

9. The CAR of claim 8, wherein the transmembrane domain(s) is selected from a transmembrane domain of CD8a including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 21 ; a transmembrane domain of CD28 including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 22; a transmembrane domain of CD16a including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 23, a transmembrane domain of 2B4 including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 24; or a transmembrane domain of NKG2D including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 25.

10. The CAR of any of claims 1 to 9, further comprising one or more intracellular signaling domains.

11. The CAR of any of claims 1 to 10, wherein the one or more intracellular signaling domain(s) is selected from a 4 IBB intracellular signaling domain, a CD28 intracellular signaling domain, a CD3^ intracellular signaling domain, aCD16A intracellular signaling domain, a yc intracellular signaling domain, a 2B4 intracellular signaling domain, a DAP10 intracellular signaling domain, a DAP12 intracellular signaling domain, and/or combinations thereof.

12. The CAR of claim 10, wherein the intracellular signaling domain(s) is selected from an intracellular domain of 41BB including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 26; an intracellular domain of CD28 including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO:27; an intracellular domain of CD3C, including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 28; an intracellular domain of 0X40 including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ TD NO: 29; an intracellular domain of CD16A including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 30; an intracellular domain of yc including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 31 ; an intracellular domain of 2B4 including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 32; an intracellular domain of DAP10 including an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 33; or an intracellular domain of DAP 12 including an amino acid sequence at least about 70%, at least about 71 %, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 34.

13. The CAR of any of claims 1 to 12, further comprising a spacer.

14. The CAR of claim 13, wherein the spacer comprises an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 35.

15. The CAR of any of claims 1 to 14, further comprising a signaling peptide.

16. The CAR of claim 15, wherein the signaling peptide comprises an amino acid sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 36 or SEQ ID NO: 37.

17. The CAR of any of claims 1 to 4 or 6 to 16, comprising from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a hinge domain, a transmembrane domain, a CD28 intracellular domain; and an 0X40 intracellular domain.

18. The CAR of any of claims 1 to 4 or 6 to 16, comprising from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, an IgGl hinge domain, a CD28 transmembrane domain, the CD28 intracellular domain, the 0X40 intracellular domain, and a CD3^ intracellular domain.

19. The CAR of any of claims 1 to 4 or 6 to 16, comprising from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a CD 16a intracellular signaling domain, a 2B4 intracellular domain, and a CD3^ intracellular domain.

20. The CAR of any of claims 1 to 4 or 6 to 16, comprising from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a yc intracellular signaling domain, a 2B4 intracellular domain, and a CD3^ intracellular domain.

21. The CAR of any of claims 1 to 4 or 6 to 16, comprising from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a CD28 intracellular signaling domain, a 2B4 intracellular domain, and a CD3^ intracellular domain.

22. The CAR of any of claims 1 to 4 or 6 to 16, comprising from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a 2B4 intracellular signaling domain, a 41BB intracellular domain, and a CD3 intracellular domain.

23. The CAR of any of claims 1 to 4 or 6 to 16, comprising from N-to-C-terminus, an extracellular domain that comprises the OSM ligand, a spacer, an IgGl hinge domain, a transmembrane domain, a 2B4 intracellular signaling domain, a DAP10 intracellular domain, a DAP 12 intracellular domain, and a CD3^ intracellular domain.

24. A nucleic acid comprising a nucleotide sequence encoding the CAR of any of claims 1 to 23.

25. The nucleic acid of claim 24, wherein the nucleotide is operably linked to a promoter.

26. An expression construct comprising the nucleic acid of claim 24 or 25.

27. The expression construct of claim 26, comprising a vector.

28. The expression construct of claim 27, wherein the vector is a retroviral vector, a lentiviral vector, or an AAV vector.

29. The expression construct of any of claims 26 to 28, further comprising a nucleotide sequence encoding a cytokine.

30. The expression construct of claim 29, wherein the cytokine is IL- 15, IL-12, IL-2, IL- 18, IL-21, or a combination thereof.

31. The expression construct of claim 30, comprising a nucleotide sequence at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 6.

32. An immune cell comprising a CAR of any of claims 1 to 23.

33. An immune cell transfected or transduced with the expression construct of any of claims 26 to 31.

34. The immune cell of claims 32 or 33, comprising at least one of cytotoxic T lymphocytes, natural killer cells, or natural killer T cells.

35. An immunotherapy composition comprising a plurality of immune cells of any of claims 32 to 34.

36. A method of treating cancer in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of the immunotherapy composition of claim 35.

37. The method of claim 36, wherein the cancer comprises a solid tumor.

38. The method of claim 36, wherein the cancer comprises at least one of osteosarcoma, melanoma, mesothelioma, breast cancer, prostate cancer, head and neck cancer, lung cancer, or brain cancer.

39. The method of claim 36, wherein the cancer comprises a hematological cancer or malignancy.

40. The method of claim 39, wherein the cancer or malignancy is acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma, marginal zone lymphoma, Acute lymphoblastic leukemia, non-Hodgkin’ s lymphoma, Hodgkin’s lymphoma, B cell malignancies, and multiple myeloma.