Macrophage cars and uses thereof

Abstract

Macrophage CARs and nucleic acids encoding the same as well as macrophages comprising the CARs are provided. Methods of use, such as for treating cancers, and pharmaceutical compositions for treating the same are further provided.

Description

[0001] Docket No. 3157.W01WO / 680.3157WO01

[0002] MACROPHAGE CARS AND USES THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 777,314, filed on March 25, 2025, the disclosure of which is incorporated by reference herein in its entirety.

[0004] SEQUENCE LISTING

[0005] This application contains a Sequence Listing electronically submitted via Patent Center to the United States Patent and Trademark Office as an XML file entitled “0680.003157W001.xml” having a size of 86,278 bytes and created on March 18, 2026. The information contained in the Sequence Listing is incorporated by reference herein.

[0006] BACKGROUND

[0007] Chimeric antigen receptors represent a significant advance in immune therapies, particularly for treating cancers. Until now, most CAR therapeutics are CAR-T cell therapeutics, which have had a demonstrable positive impact, particularly for treating hematological cancers. Using engineered immune cells to treat additional cancer types remains less advanced than T cell-based CAR therapies. In particular, CAR-based therapies have been limited in effectively treating solid cancers. Macrophages comprising CARs (CAR-Ms) are a promising technology that would benefit from new advances providing effective compositions and methods of treating solid cancers (e.g., U.S. Pat. App. Pub. No. 2024 / 0066126; CN114657143; CN113106067; and CN114854692).

[0008] Therefore, effective CAR-M therapies that target cancers or tumors, such as solid tumors, or cell surface antigens expressed on cancers or tumors, such as solid tumors, are desirable.

[0009] SUMMARY

[0010] Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0011] Formula I. R1 - R2 - R3 - R4

[0012] wherein:

[0013] R1 comprises an extracellular binding domain;

[0014] R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4);

[0015] R3 is a transmembrane domain derived from toll-like receptor 4 (TLR4); and

[0016] R4 comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4). Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:Formula II. R5 - R2 - R6 - R7

[0017] wherein:

[0018] R5 comprises an extracellular domain means for binding HER2;

[0019] R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4);

[0020] R6 is a protein means derived from toll-like receptor 4 (TLR4); and

[0021] R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0022] Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0023] Formula III. R1 - R8 - R3 - R4

[0024] wherein:

[0025] R1 comprises an extracellular binding domain;

[0026] R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R3 is a transmembrane domain derived from toll-like receptor 4 (TLR4); and

[0027] R4 comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4). Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0028] Formula IV. R5 - R8 - R6 - R7

[0029] wherein:

[0030] R5 comprises an extracellular domain means for binding HER2;

[0031] R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R6 is a protein means derived from toll-like receptor 4 (TLR4); and

[0032] R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0033] Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0034] Formula V. R9 - R2 - R6 - R7

[0035] wherein:

[0036] R9 comprises an extracellular domain means for binding ALK;

[0037] R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4);

[0038] R6 is a protein means derived from toll-like receptor 4 (TLR4); and

[0039] R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0040] Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0041] Formula VI. R9 - R8 - R6 - R7

[0042] wherein:

[0043] R9 comprises an extracellular domain means for binding ALK;

[0044] R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4);R6 is a protein means derived from toll-like receptor 4 (TLR4); and

[0045] R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0046] Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0047] Formula VII. R1 -R10 -R11 - R12

[0048] wherein:

[0049] R1 comprises an extracellular binding domain;

[0050] RIO is a hinge or spacer region derived from cluster of differentiation 28 (CD28);

[0051] R11 is a transmembrane domain derived from cluster of differentiation 28 (CD28); and R12 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0052] Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0053] Formula VIII. R5 - R13 - R14 - R15

[0054] wherein:

[0055] R5 comprises an extracellular domain means for binding HER2;

[0056] R13 is a protein means derived from cluster of differentiation 28 (CD28) for linking an extracellular means for binding HER2 and a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane;

[0057] R14 is a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane and

[0058] R15 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0059] Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0060] Formula IX. R1 - R16 - R17 - R12

[0061] wherein:

[0062] R1 comprises an extracellular binding domain;

[0063] R16 is a hinge or spacer region derived from cluster of differentiation 8 (CD8);

[0064] R17 is a transmembrane domain derived from interferon-gamma receptor 1 (IFNgRl); and R12 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0065] Provided herein are macrophage chimeric antigen receptors (CARs) of the formula:

[0066] Formula X. R9 - R18 - R19 - R15

[0067] wherein:R9 comprises an extracellular domain means for binding ALK;

[0068] R18 is a protein means derived from cluster of differentiation 8 (CD8);

[0069] R19 is a protein means derived from interferon-gamma receptor 1 (IFNgRl); and

[0070] R15 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0071] Provided herein are macrophage chimeric antigen receptors (CARs) comprising SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, or SEQ ID NO: 63.

[0072] Provided herein are vectors for engineering a macrophage, wherein the vector is a retroviral vector, comprising a nucleic acid encoding the macrophage CAR of any one of claims 1-61.

[0073] Provided herein are engineered macrophages, comprising a CAR as provided herein.

[0074] Provided herein are pharmaceutical compositions, comprising an engineered macrophage as provided here; and a pharmaceutically acceptable carrier.

[0075] Provided herein are methods of treating cancer, comprising administering to a subject with cancer: an engineered macrophage as provided here; or a pharmaceutical composition as provided herein.

[0076] BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows embodiments of a reporter cell line detecting CAR-driven activation of signaling pathways.

[0077] FIGS. 2A-2C show embodiments of a reporter cell line detecting CAR-driven activation of signaling pathways.

[0078] FIGS. 3A-3B shows embodiments of assaying CAR functionality in primary macrophages targeting tumor antigen anaplastic lymphoma kinase (ALK).

[0079] FIG. 4 shows embodiments of assaying CAR functionality in primary macrophages targeting tumor antigen anaplastic lymphoma kinase (ALK).

[0080] FIG. 5 shows embodiments of assaying CAR functionality in primary macrophages targeting tumor antigen human epidermal growth factor receptor 2 (HER2).

[0081] FIG. 6 shows embodiments of assaying CAR functionality in primary macrophages targeting tumor antigens ALK and HER2.DETAILED DESCRIPTION

[0082] The nucleic acids provided herein can be administered to subjects or patients. Herein, “administration” refers to the act of the attending physician or caregiver, prescribing the agent for administration and thereby causing the application of an agent to a subject, through ingestion, infusion, injection, or any other means, whether self-administered or administered by a clinician or other qualified caregiver. Herein, a “subject” includes both human patients and veterinary subjects, including human and non-human mammals.

[0083] Anaplastic lymphoma kinase (ALK; also known as ALK receptor tyrosine kinase; see, e.g., OMIM No. 105590) is a receptor tyrosine kinase involved in Wnt / beta-catenin signaling.

[0084] Dysregulation of ALK is associated with various cancers, including neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma. Antibodies and fragments thereof that bind ALK are known. In embodiments, antibody scFvs that bind ALK are contemplated herein. In embodiments, scFvs that bind ALK comprise SEQ ID NO: 31 (e.g., as encoded by SEQ ID NO: 32).

[0085] Used herein, an "antibody" is an immunoglobulin molecule comprising two heavy chains (HCs) and two light chains (LCs) interconnected by disulfide bonds. The amino terminal portion of each LC and HC includes a variable region of about 100-120 amino acids primarily responsible for antigen recognition via the complementary determining region (CDRs) contained therein. The CDRs are interspersed with well-known and generally conserved regions among and between species (e.g., mouse and human), which are termed framework regions (FRs).

[0086] Further disclosed herein are antibody fragments, such as well-characterized Fabs (e.g., Fab, Fab', F(ab')2,), Fvs (the variable region of the light chain and the variable region of the heavy chain expressed as two chains), and single-chain fragments (e.g., single-chain variable region fragments, scFv, and single chain Fabs, scFab), which also bind to a target (e.g., ALK or HER2). Methods of making these fragments are routine (see, e.g., Antibody Engineering: Methods and Protocols (Damien Nevoltris and Patrick Chames eds., 3d ed. 2018)).

[0087] In embodiments, antibody fragments provided herein comprise, consist essentially of, or consist of scFvs. In embodiments, antibody fragments provided herein comprise, consist essentially of, or consist of scFvs that bind ALK or HER2. In embodiments, antibody fragments provided herein comprise, consist essentially of, or consist of scFvs that bind ALK. In embodiments, antibodyfragments provided herein comprise scFvs that bind ALK. In embodiments, antibody fragments provided herein consist essentially of scFvs that bind ALK. In embodiments, antibody fragments provided herein consist of scFvs that bind ALK. In embodiments, antibody fragments provided herein comprise, consist essentially of, or consist of scFvs that bind HER2. In embodiments, antibody fragments provided herein comprise scFvs that bind HERZ. In embodiments, antibody fragments provided herein consist essentially of scFvs that bind HER2. In embodiments, antibody fragments provided herein consist of scFvs that bind HER2.

[0088] “Chimeric antigen receptors” (CARs) are receptor proteins that have been engineered for expression on an immune cell and to target a specific antigen (“target antigen”) as well as activate the immune cell. CARs are used in therapies, such as immune cell therapy, including T and NK cell therapy. Virus- or RNA-encoding CARs can be used for in vivo generation of CAR effector cells. CARs can be engineered into allogeneic immune cells (i.e., immune cells from a donor are engineered) or autologous immune cells (i.e., immune cells from a patient or subject re-introduced after engineering). CARs typically include (1) extracellular binding domain (e.g., including antibody or fragment thereof, such as heavy chain variable region(s)), which is an antigen-binding part of the CAR that is essentially free of the transmembrane and cytoplasm, (2) hinge and / or spacer region, which is an amino acid sequence in the CAR that is positioned between the extracellular antigenbinding domain and transmembrane domain, (3) transmembrane domain, which is a part of the CAR that spans from the extracellular side to the intracellular side of a cell membrane, and (4) an intracellular domain, which is the intracellular a part of the CAR and includes signaling domain (also known as an activating domain) and, optionally, costimulatory domain. In embodiments, a CAR can include an extracellular signal peptide (also known as a leader or leading peptide) N-terminal to an antibody or antibody fragment in an extracellular binding domain.

[0089] Cluster of Differentiation 28 (CD28; also known as Tp44; e.g., OMIM No. 186760) is a T-cell-specific surface glycoprotein that supports T-cell activation. CD28 enhances the production of various interleukins, including IL-6. Dysregulation of CD28 has been observed in several cancers, including melanoma, non-small cell lung cancer, head and neck squamous cell carcinoma, gastric cancer, ovarian cancer, cervical cancer, glioblastoma, and breast cancer, often associated with T cell dysfunction and exhaustion. In embodiments, protein domains derived from CD28 are contemplated herein. In embodiments, protein domains derived from CD28 are contemplated as a hinge (also known as a spacer) or transmembrane domain. In embodiments, protein domains derived from CD28comprise SEQ ID NO: 15 (e.g., as encoded by SEQ ID NO: 16). In embodiments, protein domains derived from CD28 comprise SEQ ID NO: 17 (e.g., as encoded by SEQ ID NO: 18).

[0090] Cluster of Differentiation 8 (CD8; also known as p32, OKT8 T-cell antigen; e.g., OMIM No.

[0091] 186910) is a transmembrane glycoprotein involved in T-cell activity. CD8 is expressed on cytotoxic T cells (i.e., CD8+ T cells) and participates in antigen recognition. Dysregulation of CD8+ T cells has been observed in several cancers, including melanoma, non-small cell lung cancer, head and neck squamous cell carcinoma, gastric cancer, and ovarian cancer, leading to T-cell exhaustion, immune evasion, and tumor progression. In embodiments, protein domains derived from CD8 are contemplated herein. In embodiments, protein domains derived from CD8 are contemplated as a signal peptide or hinge (also known as a spacer) domain. In embodiments, protein domains derived from CD8 comprise SEQ ID NO: 35 (e.g., as encoded by SEQ ID NO: 36). In embodiments, protein domains derived from CD8 comprise SEQ ID NO: 23 (e.g., as encoded by SEQ ID NO: 24).

[0092] Granulocyte-Macrophage Colony-Stimulating Factor Receptor (GM-CSFR; e.g., OMIM No.

[0093] 306250) is a protein that binds to granulocyte-macrophage colony-stimulating factor (GM-CSF), a cytokine that controls the production of white blood cells. GM-CSFR is primarily located on neutrophils, eosinophils, and monocytes / macrophages and is known to activate SRC family kinases and JAK-STAT, PI3K, and MAPK signaling pathways. Abnormal expression or activity of GM-CSFR can lead to dysregulation of immune responses and contribute to cancer progression. In embodiments, protein domains derived from GM-CSFR are contemplated herein. In embodiments, protein domains derived from GM-CSFR are contemplated as a signal peptide. In embodiments, protein domains derived from GM-CSFR comprise SEQ ID NO: 33 (e.g., as encoded by SEQ ID NO: 34).

[0094] Human epidermal growth factor receptor 2 (HER2; also known as ERB-B2 receptor tyrosine kinase 2 (ERBB2), V-ERB-B2 avian erythroblastic leukemia viral oncogene homolog 2, oncogene ERBB2, oncogene NGL, neuroblastoma- or glioblastoma-derived (NGL-NEU), tyrosine kinase-type cell surface receptor HER2 (TKR1 HER2); see, e.g., OMIM No. 164870) is a receptor tyrosine kinase that promotes cell proliferation and suppresses apoptosis. Dysregulation of HER2 is associated with various cancers, including breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, and cervical cancer. Antibodies and fragments thereof that bind HER2 are known. In embodiments, antibody scFvs that bind HER2 are contemplated herein. Inembodiments, a scFv that binds HER2 comprises SEQ ID NO: 29 (e.g., as encoded by SEQ ID NO: 30).

[0095] Immunoglobulin kappa (IgK; e.g., OMIM No. 147200) is a type of light chain found in immunoglobulins, which are glycoproteins produced by B cells and secreted by plasma cells as antibodies. The IgK light chain, together with the heavy chain, forms the structure of an antibody, a component in the adaptive immune response. The IgK light chain contributes to antigen-binding specificity of antibodies. In embodiments, protein domains derived from IgK are contemplated herein. In embodiments, protein domains derived from IgK are contemplated as a signal peptide. In embodiments, protein domains derived from IgK comprise SEQ ID NO: 27 (e.g., as encoded by SEQ ID NO: 28).

[0096] Interferon-gamma receptor 1 (IFNGR1; also known as IFNGR, IMD27A, IMD27B, antiviral protein (AVP) type II, immune interferon receptor 1, CD119 antigen, and CD119; see, e.g., OMIM No. 107470) is a transmembrane receptor involved in interferon gamma signaling and is the ligandbinding chain (alpha) of the interferon gamma receptor heterodimer. Various phenotypes are associated with altered IFNGR1 expression, including phenotypes related to cancer or infectious disease. IFNGR1 protein and nucleic acid sequences are known in the art, for example human protein sequences, such as GenBank accession no. NP_000407.1, and human nucleic acid sequences, such as GenBank accession no. NM_000416.3. In embodiments, protein domains derived from IFNGR1 are contemplated herein. In embodiments, protein domains derived from IFNGR1 are contemplated as a transmembrane domain or intracellular signaling domain. In embodiments, protein domains derived from IFNGR1 comprise SEQ ID NO: 25 (e.g., as encoded by SEQ ID NO: 26). In embodiments, protein domains derived from IFNGR1 comprise SEQ ID NO: 37 (e.g., as encoded by SEQ ID NO: 38).

[0097] Interferon-gamma receptor 2 (IFNGR2; also known as interferon-gamma transducer 1 (IFNGT1), accessory factor for interferon-gamma receptor; see, e.g., OMIM No. 147569) is a transmembrane receptor involved in interferon gamma signaling and is the non-ligand-binding chain (beta) of the interferon gamma receptor heterodimer. Various phenotypes are associated with altered IFNGR2 expression, including phenotypes related to cancer or infectious disease. IFNGR2 protein and nucleic acid sequences are known in the art, for example human protein sequences, such as GenBank accession no. AAH03624.1, and human nucleic acid sequences, such as GenBank accession no. NM_001329128.2. In embodiments, protein domains derived from IFNGR2 are contemplated herein. In embodiments, protein domains derived from IFNGR2 are contemplated asan intracellular signaling domain. In embodiments, protein domains derived from IFNGR2 comprise SEQ ID NO: 39 (e.g., as encoded by SEQ ID NO: 40).

[0098] Used herein, a linkage is identified as a

[0099]

[0100] located between various sections of a CAR (e.g., between the R groups of the formulas disclosed herein), read left to right as the N-terminal side to the C-terminal side of the CAR. A linkage can be a direct peptide bond or a linker (also known as a spacer). In embodiments, the linkage is a linker (also known as a spacer), which is a chain of amino acids linking together two consecutive functional domains through direct peptide bonds. In embodiments, a linker can be a combination of glycine and serine amino acids (also known as a ‘GS linker’). Various GS linkers are possible. Other amino acids may be present in a GS linker, so long as glycine and serine residues comprise 50% to 100% of the amino acids in the GS linker sequence. In embodiments, a GS linker is a sequence of 3 or 4 glycines followed by a serine (i.e., GGGS, SEQ ID NO: 42, or GGGGS, SEQ ID NO: 41). In embodiments, a GS linker is a sequence of 4 glycines followed by a serine. In embodiments, a GS linker has repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 3 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGS GGGGS GGGGS (SEQ ID NO: 43). In embodiments, a GS linker has 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 45). In embodiments, a linker is a protein tag (e.g., HIS tag or FLAG tag). In embodiments, a linker is a FLAG tag (e.g., SEQ ID NO: 47). In embodiments, a protein tag linker is optional.

[0101] “Macrophages,” as used herein, are a type of white blood cell of the innate immune system that engulfs and digests pathogens (e.g., cancer cells, microbes, cellular debris, and foreign substances), a process called “phagocytosis” that defends the host against infection and injury. Macrophages, such as Ml macrophages, can also help initiate adaptive immunity by recruiting other immune cells (e.g., lymphocytes) and participating in antigen presentation to T cells. Macrophages, such as M2 macrophages, also have an anti-inflammatory role by decreasing immune reactions through the release of cytokines. Macrophage CAR (also known as CAR-M) is typically employed in an Ml model to enhance an immune response against cancer cells. Macrophages and other immune cells are derived from monocytes, which are generated from long-term hematopoietic stem cells (LT-HSCs), and can differentiate into macrophages or dendritic cells. Ml macrophages can be produced through monocyte differentiation stimulated by the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Other methods of producing macrophages, such as CAR-M (e.g., an Ml macrophage transduced with CAR) are well-known in the art (e.g., Sloas et al., 2021,Engineered CAR-Macrophages as Adoptive Immunotherapies for Solid Tumors. Front. Immunol.

[0102] 12:783305. doi: 10.3389 / fimmu.2021.783305).

[0103] The pharmaceutically acceptable carriers of use are conventional (e.g., as described in Remington, The Science and Practice of Pharmacy, 22nd Edition, Loyd V., ed., Pharmaceutical Press, 2012). In general, the nature of the carrier will depend on the mode of administration. For instance, parenteral formulations typically comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids, such as water, physiological saline, balanced salt solutions or the like as a vehicle. Pharmaceutical compositions can additionally include minor amounts of nontoxic auxiliary substances for stability.

[0104] In embodiments, the carrier may be sterile and / or suspended or otherwise contained in a unit dosage form, including one or more measured doses of the composition suitable for administration to a subject of an effective amount of the nucleic acids disclosed herein. Medications for use in treatment may also be included in embodiments. In embodiments, the unit dosage form may be in a sealed vial that contains sterile contents or a syringe for injection into a subject, lyophilized for subsequent solubilization and administration, or in a solid or controlled release dosage. Toll-like receptor 4 (TLR4; also known as homolog of toll; e.g., OMIM No. 603030) is a transmembrane protein that plays a role in pathogen recognition and activation of innate immunity. TLR4 is expressed in immune cells of myeloid origin, including monocytes, macrophages, and dendritic cells. Ligands for TLR4 are lipopolysaccharides, the significant components of the outer membrane of gram-negative bacteria. Dysregulated TLR4 signaling has been implicated in the development and progression of various cancers, including colorectal cancer, breast cancer, hepatocellular carcinoma, and colitis-associated colon cancer. In embodiments, protein domains derived from TLR4 are contemplated herein. In embodiments, protein domains derived from TLR4 are contemplated as a transmembrane domain or intracellular signaling domain. In embodiments, protein domains derived from TLR4 comprise SEQ ID NO: 9 (e.g., as encoded by SEQ ID NO: 10). In embodiments, protein domains derived from TLR4 comprise SEQ ID NO: 11 (e.g., as encoded by SEQ ID NO: 12). In embodiments, truncated protein domains derived from TLR4 are contemplated as a hinge (also known as a spacer) domain. In embodiments, truncated protein domains derived from TLR4 comprise SEQ ID NO: 7 (e.g., as encoded by SEQ ID NO: 8). In embodiments, short truncated protein domains derived from TLR4 are contemplated as a hinge (also known as a spacer) domain. In embodiments, short truncated protein domains derived from TLR4 comprise SEQ ID NO: 13 (e.g., as encoded by SEQ ID NO: 14). The nucleic acids, engineered macrophages, or pharmaceuticalcompositions thereof disclosed herein can be used in therapy. In embodiments, the nucleic acids disclosed herein can be used to treat, prevent (such as through prophylactic treatment), or ameliorate a cancer, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma. Herein, “preventing” a disease refers to inhibiting the full development of a disease, such as cancer, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma. “Treating” refers to a therapeutic intervention that ameliorates a sign or symptom of a disease or pathological condition after it has begun to develop, such as a reduction in tumor burden or a decrease in the number or size of metastases.

[0105] “Ameliorating” refers to the reduction in the number or severity of signs or symptoms of a disease, such as cancer, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma. A "prophylactic" treatment is a treatment administered to a subject who does not exhibit signs of a disease or exhibits only early signs for the purpose of decreasing the risk of developing a pathology, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma.

[0106] Herein, an “effective amount” is a quantity sufficient to achieve a desired effect in a subject. For instance, this can be the amount necessary to prevent, treat, or ameliorate a disease, for example, inhibiting or suppressing cancer, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer,cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma. In embodiments, an effective amount is the amount necessary to eliminate, reduce the size, or prevent metastasis of cancer or a tumor, such as in breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma. Efficacy is first evident in the cellular response, for which various in vitro and cell-based assays are well-known to measure. Kristina V. Kitaeva et al., Cell Culture Based In Vitro Test Systems for Anti cancer Drug Screening, 8 Front. Bioeng. Biotechnol. 322(2020)). In embodiments, an effective amount is the amount necessary to significantly inhibit or reduce cancer cell proliferation or migration, invasion, or adhesion. A cellular response manifests as significantly reduced tumor size, reduced or inhibited disease progression, and improvement in survival in a subject or patient. More particularly, an effective amount provides improvement in important cancer endpoints, Overall Survival (OS), Disease-Free Survival (DFS), Objective Response Rate, Complete Response Rate or Progression Free Survival (PFS). See Dept, of Health and Human Services, Food and Drug Admin, Clinical Trial Endpoints for the Approval of Cancer Drugs and Biologies: Guidance for Industry (2018); E.A. Eisenhauer et al., New Response Evaluation Criteria in Solid Tumours: Revised RECIST Guideline (Version 1.1), 45 Eur. J. Cancer 228 (2009).

[0107] “Vector,” as used herein, is an entity containing a nucleic acid molecule (such as a DNA or RNA molecule) comprising the coding sequence of a protein of interest and can express the coding sequence. In embodiments, vectors herein bear a promoter(s) that is operationally linked to the coding sequence of a protein of interest (e.g., the CAR sequences disclosed herein). Non-limiting examples include a naked or packaged (lipid and / or protein) DNA, a naked or packaged RNA, a subcomponent of a virus or bacterium or other microorganism that may be replication incompetent, or a virus or bacterium or other microorganism that may be replication-competent. A vector is sometimes referred to as a construct. Recombinant DNA vectors are vectors having recombinant DNA. A vector can include nucleic acid sequences that permit it to replicate in a host cell, such as an origin of replication. A vector can also include one or more selectable marker genes and other genetic elements. Viral vectors are recombinant nucleic acid vectors having at least some nucleicacid sequences derived from one or more viruses. In embodiments, a viral vector comprises a retroviral vector. In some embodiments, a viral vector comprises a lentivirus vector. In some embodiments, a viral vector comprises a nucleic acid molecule encoding a disclosed CAR.

[0108] COMPOSITIONS

[0109] Provided in the present disclosure are chimeric antigen receptors (CARs) and nucleic acids for expressing a CAR. Nomenclature used herein to identify the composition elements is as follows:

[0110] R1 comprises an extracellular binding domain

[0111] R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4)

[0112] R3 is a transmembrane domain derived from toll-like receptor 4 (TLR4)

[0113] R4 comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4) R5 comprises an extracellular domain means for binding HER2

[0114] R6 is a protein means derived from toll-like receptor 4 (TLR4) for traversing a cell membrane

[0115] R7 is an intracellular means derived from toll-like receptor 4 (TLR4) for activating a macrophage

[0116] R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4) R9 comprises an extracellular domain means for binding ALK

[0117] RIO is a hinge or spacer region derived from cluster of differentiation 28 (CD28) Rll is a transmembrane domain derived from cluster of differentiation 28 (CD28) Rll comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2)

[0118] R13 is a protein means derived from cluster of differentiation 28 (CD28) for linking an extracellular means for binding HER2 and a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane

[0119] R14 is a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane

[0120] R15 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) for intracellular signaling R16 is a hinge or spacer region derived from cluster of differentiation 8 (CD8)

[0121] R17 is a transmembrane domain derived from interferon-gamma receptor 1 (IFNgRl)R18 is a protein means derived from cluster of differentiation 8 (CD8) for linking an extracellular means for binding ALK and a protein means derived from interferongamma receptor 1 (IFNgRl) for traversing a cell membrane

[0122] R19 is a protein means derived from interferon-gamma receptor 1 (IFNgRl) for traversing a cell membrane

[0123] R20 is a signal peptide derived from immunoglobulin K (IgK)

[0124] R21 is an antibody fragment

[0125] R22 is a signal peptide derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR)

[0126] R23 is a protein means derived from immunoglobulin K (IgK) for directing translocation of the CAR

[0127] R24 is a protein means derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR) for directing translocation of the CAR

[0128] R25 an antibody fragment means for binding HER2

[0129] R26 an antibody fragment means for binding ALK

[0130] R27 is a signal peptide derived from cluster of differentiation 8 (CD8

[0131] R28 is a protein means derived from cluster of differentiation 8 (CD8) for directing translocation of the CAR

[0132] R29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl)

[0133] R30 comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2)

[0134] R31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) for intracellular signaling

[0135] R32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2) for intracellular signaling

[0136] Formula I R1 - R2 - R3 -R4

[0137] Formula II R5 - R2 - R6 - R7

[0138] Formula III R1 - R8 - R3 - R4

[0139] Formula IV R5 - R8 - R6 - R7

[0140] Formula V R9 - R2 - R6 -R7Formula VI R9 - R8 - R6 - R7

[0141] Formula VII R1 - RIO - R11 - R12

[0142] Formula VIII R5 - R13 - R14 - R15

[0143] Formula IX R1 - R16 - R17 - R12

[0144] Formula X R9 - R18 - R19 - R15

[0145] Formula XI R20 -R21

[0146] Formula XII R23 - R25

[0147] Formula XIIIR22 -R21

[0148] Formula XIVR24 -R26

[0149] Formula XV R27 -R21

[0150] Formula XVIR28 -R26

[0151] Formula XVII R29 - R30

[0152] Formula XVIII R30 - R29

[0153] Formula XIX R31 - R32

[0154] Formula XX R32 -R31

[0155] SEQ ID NOs: 1-63 include nucleic acid and amino acid sequences of components that may be included in the CARs of the present disclosure.

[0156] SEQ ID NOs: 1-6 are examples of an embodiment amino acid and nucleic acid sequences of extracellular binding domains.

[0157] SEQ ID NO: 7 is an embodiment amino acid sequence of a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). SEQ ID NO: 8 is an embodiment nucleic acid sequence of a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). SEQ ID NO: 9 is an embodiment amino acid sequence of a transmembrane domain derived from toll-like receptor 4 (TLR4). SEQ ID NO: 10 is an embodiment nucleic acid sequence of a transmembrane domain derived from toll-like receptor 4 (TLR4). SEQ ID NO: 11 is an embodiment amino acid sequence of an intracellular signaling domain derived from toll-like receptor 4 (TLR4). SEQ ID NO: 12 is an embodiment nucleic acid sequence of an intracellular signaling domain derived from toll-like receptor 4 (TLR4). SEQ ID NO: 13 is an embodiment amino acid sequence of a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). SEQ ID NO: 14 is an embodiment nucleic acid sequence of a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4).SEQ ID NO: 15 is an embodiment amino acid sequence of a hinge or spacer region derived from cluster of differentiation 28 (CD28). SEQ ID NO: 16 is an embodiment nucleic acid sequence of a hinge or spacer region derived from cluster of differentiation 28 (CD28). SEQ ID NO: 17 is an embodiment amino acid sequence of a transmembrane domain derived from cluster of differentiation 28 (CD28). SEQ ID NO: 18 is an embodiment nucleic acid sequence of a transmembrane domain derived from cluster of differentiation 28 (CD28).

[0158] SEQ ID NO: 19 is an embodiment amino acid sequence of an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2). SEQ ID NO: 20 is an embodiment nucleic acid sequence of an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2). SEQ ID NO: 21 is an embodiment amino acid sequence of an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2). SEQ ID NO: 22 is an embodiment nucleic acid sequence of an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0159] SEQ ID NO: 23 is an embodiment amino acid sequence of a hinge or spacer region derived from cluster of differentiation 8 (CD8). SEQ ID NO: 24 is an embodiment nucleic acid sequence of a hinge or spacer region derived from cluster of differentiation 8 (CD8).

[0160] SEQ ID NO: 25 is an embodiment amino acid sequence of a transmembrane domain derived from interferon-gamma receptor 1 (IFNgRl). SEQ ID NO: 26 is an embodiment nucleic acid sequence of a transmembrane domain derived from interferon-gamma receptor 1 (IFNgRl).

[0161] SEQ ID NO: 27 is an embodiment amino acid sequence of a signal peptide derived from immunoglobulin K (IgK). SEQ ID NO: 28 is an embodiment nucleic acid sequence of a signal peptide derived from immunoglobulin K (IgK).

[0162] SEQ ID NO: 29 is an embodiment amino acid sequence of an antibody fragment. SEQ ID NO: 30 is an embodiment nucleic acid sequence of an antibody fragment. SEQ ID NO: 31 is an embodiment amino acid sequence of an antibody fragment. SEQ ID NO: 32 is an embodiment nucleic acid sequence of an antibody fragment.

[0163] SEQ ID NO: 33 is an embodiment amino acid sequence of a signal peptide derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR). SEQ ID NO: 34 is an embodiment nucleic acid sequence of a signal peptide derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR).SEQ ID NO: 35 is an embodiment amino acid sequence of a signal peptide derived from cluster of differentiation 8 (CD8). SEQ ID NO: 36 is an embodiment nucleic acid sequence of a signal peptide derived from cluster of differentiation 8 (CD8).

[0164] SEQ ID NO: 37 is an embodiment amino acid sequence of an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl). SEQ ID NO: 38 is an embodiment nucleic acid sequence of an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl). SEQ ID NO: 39 is an embodiment amino acid sequence of an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2). SEQ ID NO: 40 is an embodiment nucleic acid sequence of an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2).

[0165] SEQ ID NOs: 41-46 are examples of embodiment amino acid and nucleic acid sequences of GS linkers. SEQ ID NOs: 47 and 48 an embodiment amino acid sequence of FLAG tags.

[0166] SEQ ID NOs: 49-64 are examples of embodiment amino acid and nucleic acid sequences of CARs as disclosed herein.

[0167] In embodiments, the CAR is of Formula I, or the nucleic acid comprises a nucleic acid sequence encoding a CAR of the formula:

[0168] Formula I. R1 - R2 - R3 - R4

[0169] wherein:

[0170] R1 comprises an extracellular binding domain;

[0171] R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4);

[0172] R3 is a transmembrane domain derived from toll-like receptor 4 (TLR4); and

[0173] R4 comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4).

[0174] In embodiments, R1 of Formula I comprises an extracellular binding domain. In embodiments, R1 of Formula I comprises an antibody or antibody fragment. In embodiments, R1 of Formula I comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R1 of Formula I comprises an scFv. In embodiments, R1 of Formula I comprises an scFv that binds HER2 or ALK. In embodiments, R1 of Formula I comprises SEQ ID NO: 1 or SEQ ID NO: 3.

[0175] In embodiments, R1 of Formula I is an scFv. In embodiments, R1 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 1. In embodiments, R1 is an amino acid sequence comprising SEQ ID NO: 1 or SEQ ID NO: 3. In embodiments, R1 is an aminoacid sequence consisting essentially of SEQ ID NO: 1 or SEQ ID NO: 3. In embodiments, R1 is an amino acid sequence consisting of SEQ ID NO: 1 or SEQ ID NO: 3.

[0176] In embodiments, R2 of Formula I is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). In embodiments, R1 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 7. In embodiments, R1 is an amino acid sequence comprising SEQ ID NO: 7. In embodiments, R1 is an amino acid sequence consisting essentially of SEQ ID NO: 7. In embodiments, R1 is an amino acid sequence consisting of SEQ ID NO: 7.

[0177] In embodiments, R3 of Formula I is a transmembrane domain derived from toll-like receptor 4 (TLR4). In embodiments, R3 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 9. In embodiments, R3 is an amino acid sequence comprising SEQ ID NO: 9. In embodiments, R3 is an amino acid sequence consisting essentially of SEQ ID NO: 9. In embodiments, R3 is an amino acid sequence consisting of SEQ ID NO: 9.

[0178] In embodiments, R4 of Formula I comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4). In embodiments, R4 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 11. In embodiments, R4 is an amino acid sequence comprising SEQ ID NO: 11. In embodiments, R4 is an amino acid sequence consisting essentially of SEQ ID NO: 11. In embodiments, R4 is an amino acid sequence consisting of SEQ ID NO: 11.

[0179] In embodiments, R1 of Formula I comprises an extracellular binding domain of the formula: Formula XI. R20 - R21

[0180] wherein:

[0181] R20 is a signal peptide derived from immunoglobulin K (IgK); and

[0182] R21 is an antibody fragment.

[0183] In embodiments, R20 of Formula XI is a signal peptide derived from immunoglobulin K (IgK). In embodiments, R20 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence comprising SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence consisting essentially of SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence consisting of SEQ ID NO: 27.

[0184] In embodiments, R21 of Formula XI is an antibody fragment. In embodiments, R21 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 29. In embodiments, R21 is an amino acid sequence comprising SEQ ID NO: 29. In embodiments, R21 isan amino acid sequence consisting essentially of SEQ ID NO: 29. In embodiments, R21 is an amino acid sequence consisting of SEQ ID NO: 29.

[0185] In embodiments, R1 of Formula I comprises an extracellular binding domain of the formula: Formula XIII. R22 -R21

[0186] wherein:

[0187] R22 is a signal peptide derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR); and

[0188] R21 is an antibody fragment.

[0189] In embodiments, R22 of Formula XIII is a signal peptide derived from granulocytemacrophage colony-stimulating factor receptor (GM-CSFR). In embodiments, R22 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 33. In embodiments, R22 is an amino acid sequence comprising SEQ ID NO: 33. In embodiments, R22 is an amino acid sequence consisting essentially of SEQ ID NO: 33. In embodiments, R22 is an amino acid sequence consisting of SEQ ID NO: 33.

[0190] In embodiments, R21 of Formula XIII is an antibody fragment. In embodiments, R21 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence comprising SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence consisting essentially of SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence consisting of SEQ ID NO: 31.

[0191] In embodiments, the CAR of Formula I is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 49. In embodiments, Formula I is an amino acid sequence comprising SEQ ID NO: 49. In embodiments, Formula I is an amino acid sequence consisting essentially of SEQ ID NO: 49. In embodiments, Formula I is an amino acid sequence consisting of SEQ ID NO: 49.

[0192] In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula:

[0193] Formula II. R5 - R2 - R6 - R7

[0194] wherein:

[0195] R5 comprises an extracellular domain means for binding HER2;

[0196] R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4);

[0197] R6 is a protein means derived from toll-like receptor 4 (TLR4); and

[0198] R7 is an intracellular means derived from toll-like receptor 4 (TLR4).In embodiments, R5 of Formula II comprises an extracellular domain means for binding HER2. In embodiments, R5 of Formula II comprises an antibody or antibody fragment. In embodiments, R5 of Formula II comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R5 of Formula II comprises an scFv. In embodiments, R5 of Formula II comprises an scFv that binds HER2. In embodiments, R5 of Formula II comprises SEQ ID NO: 1.

[0199] In embodiments, R5 of Formula II is an scFv. In embodiments, R5 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence comprising SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence consisting essentially of SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence consisting of SEQ ID NO: 1.

[0200] In embodiments, R2 of Formula II is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). In embodiments, R2 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 7. In embodiments, R2 is an amino acid sequence comprising SEQ ID NO: 7. In embodiments, R2 is an amino acid sequence consisting essentially of SEQ ID NO: 7. In embodiments, R2 is an amino acid sequence consisting of SEQ ID NO: 7.

[0201] In embodiments, R6 of Formula II is a protein means derived from toll-like receptor 4 (TLR4). In embodiments, R6 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence comprising SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence consisting essentially of SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence consisting of SEQ ID NO: 9.

[0202] In embodiments, R7 of Formula II is an intracellular means derived from toll-like receptor 4 (TLR4). In embodiments, R7 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence comprising SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence consisting essentially of SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence consisting of SEQ ID NO: 11.

[0203] In embodiments, R5 of Formula II comprises an extracellular domain means for binding HER.2 of the formula:

[0204] Formula XII. R23 - R25

[0205] wherein:R23 is a protein means derived from immunoglobulin K (IgK); and

[0206] R25 an antibody fragment means for binding HERZ.

[0207] In embodiments, R23 of Formula XII is a protein means derived from immunoglobulin K (IgK). In embodiments, R23 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence comprising SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence consisting essentially of SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence consisting of SEQ ID NO: 27.

[0208] In embodiments, R25 of Formula XII an antibody fragment means for binding HER2. In embodiments, R25 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence comprising SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence consisting essentially of SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence consisting of SEQ ID NO: 29.

[0209] In embodiments, the CAR of Formula II is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 49. In embodiments, Formula II is an amino acid sequence comprising SEQ ID NO: 49. In embodiments, Formula II is an amino acid sequence consisting essentially of SEQ ID NO: 49. In embodiments, Formula II is an amino acid sequence consisting of SEQ ID NO: 49.

[0210] In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula:

[0211] Formula III. R1 - R8 - R3 - R4

[0212] wherein:

[0213] R1 comprises an extracellular binding domain;

[0214] R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R3 is a transmembrane domain derived from toll-like receptor 4 (TLR4); and

[0215] R4 comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4). In embodiments, R1 of Formula III comprises an extracellular binding domain. In embodiments, R1 of Formula III comprises an antibody or antibody fragment. In embodiments, R1 of Formula III comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R1 of Formula III comprises an scFv. In embodiments, R1 of Formula III comprises an scFv that binds HER2 or ALK. In embodiments, R1 of Formula III comprises SEQ ID NO: 1 or SEQ ID NO: 3.In embodiments, R1 of Formula III is an scFv. In embodiments, R1 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 1 or SEQ ID NO: 3. In embodiments, R1 is an amino acid sequence comprising SEQ ID NO: 1 or SEQ ID NO: 3. In embodiments, R1 is an amino acid sequence consisting essentially of SEQ ID NO: 1 or SEQ ID NO: 3. In embodiments, R1 is an amino acid sequence consisting of SEQ ID NO: 1 or SEQ ID NO: 3.

[0216] In embodiments, R8 of Formula III is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). In embodiments, R8 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence comprising SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence consisting essentially of SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence consisting of SEQ ID NO: 13.

[0217] In embodiments, R3 of Formula III is a transmembrane domain derived from toll-like receptor 4 (TLR4). In embodiments, R3 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 9. In embodiments, R3 is an amino acid sequence comprising SEQ ID NO: 9. In embodiments, R3 is an amino acid sequence consisting essentially of SEQ ID NO: 9. In embodiments, R3 is an amino acid sequence consisting of SEQ ID NO: 9.

[0218] In embodiments, R4 of Formula III comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4). In embodiments, R4 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 11. In embodiments, R4 is an amino acid sequence comprising SEQ ID NO: 11. In embodiments, R4 is an amino acid sequence consisting essentially of SEQ ID NO: 11. In embodiments, R4 is an amino acid sequence consisting of SEQ ID NO: 11.

[0219] In embodiments, R1 of Formula III comprises an extracellular binding domain of the formula:

[0220] Formula XI. R20 - R21

[0221] wherein:

[0222] R20 is a signal peptide derived from immunoglobulin K (IgK); and

[0223] R21 is an antibody fragment.

[0224] In embodiments, R20 of Formula XI is a signal peptide derived from immunoglobulin K (IgK). In embodiments, R20 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence comprising SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence consisting essentially of SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence consisting of SEQ ID NO: 27.In embodiments, R21 of Formula XI is an antibody fragment. In embodiments, R21 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 29. In embodiments, R21 is an amino acid sequence comprising SEQ ID NO: 29. In embodiments, R21 is an amino acid sequence consisting essentially of SEQ ID NO: 29. In embodiments, R21 is an amino acid sequence consisting of SEQ ID NO: 29.

[0225] In embodiments, R1 of Formula III comprises an extracellular binding domain of the formula:

[0226] Formula XIII. R22 -R21

[0227] wherein:

[0228] R22 is a signal peptide derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR); and

[0229] R21 is an antibody fragment.

[0230] In embodiments, R22 of Formula XIII is a signal peptide derived from granulocytemacrophage colony-stimulating factor receptor (GM-CSFR). In embodiments, R22 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 33. In embodiments, R22 is an amino acid sequence comprising SEQ ID NO: 33. In embodiments, R22 is an amino acid sequence consisting essentially of SEQ ID NO: 33. In embodiments, R22 is an amino acid sequence consisting of SEQ ID NO: 33.

[0231] In embodiments, R21 of Formula XIII is an antibody fragment. In embodiments, R21 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence comprising SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence consisting essentially of SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence consisting of SEQ ID NO: 31.

[0232] In embodiments, the CAR of Formula III is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 51 or SEQ ID NO: 59. In embodiments, Formula III is an amino acid sequence comprising SEQ ID NO: 51 or SEQ ID NO: 59. In embodiments, Formula III is an amino acid sequence consisting essentially of SEQ ID NO: 51 or SEQ ID NO: 59. In embodiments, Formula III is an amino acid sequence consisting of SEQ ID NO: 51 or SEQ ID NO: 59.In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula

[0233] Formula IV. R5 - R8 - R6 - R7

[0234] wherein:

[0235] R5 comprises an extracellular domain means for binding HER2;

[0236] R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R6 is a protein means derived from toll-like receptor 4 (TLR4); and

[0237] R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0238] In embodiments, R5 of Formula IV comprises an extracellular domain means for binding HERZ. In embodiments, R5 of Formula IV comprises an antibody or antibody fragment. In embodiments, R5 of Formula IV comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R5 of Formula IV comprises an scFv. In embodiments, R5 of Formula IV comprises an scFv that binds HERZ. In embodiments, R5 of Formula IV comprises SEQ ID NO: 1.

[0239] In embodiments, R5 of Formula IV is an scFv. In embodiments, R5 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence comprising SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence consisting essentially of SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence consisting of SEQ ID NO: 1.

[0240] In embodiments, R8 of Formula IV is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). In embodiments, R8 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence comprising SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence consisting essentially of SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence consisting of SEQ ID NO: 13.

[0241] In embodiments, R6 of Formula IV is a protein means derived from toll-like receptor 4 (TLR4). In embodiments, R6 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence comprising SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence consisting essentially of SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence consisting of SEQ ID NO: 9.In embodiments, R7 of Formula IV is an intracellular means derived from toll-like receptor 4 (TLR4). In embodiments, R7 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence comprising SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence consisting essentially of SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence consisting of SEQ ID NO: 11.

[0242] In embodiments, R5 of Formula IV comprises an extracellular domain means for binding HER.2 of the formula:

[0243] Formula XII. R23 - R25

[0244] wherein:

[0245] R23 is a protein means derived from immunoglobulin K (IgK); and

[0246] R25 an antibody fragment means for binding HER2.

[0247] In embodiments, R23 of Formula XII is a protein means derived from immunoglobulin K (IgK). In embodiments, R23 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence comprising SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence consisting essentially of SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence consisting of SEQ ID NO: 27.

[0248] In embodiments, R25 of Formula XII an antibody fragment means for binding HER2. In embodiments, R25 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence comprising SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence consisting essentially of SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence consisting of SEQ ID NO: 29.

[0249] In embodiments, the CAR of Formula IV is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 51. In embodiments, Formula IV is an amino acid sequence comprising SEQ ID NO: 51. In embodiments, Formula IV is an amino acid sequence consisting essentially of SEQ ID NO: 51. In embodiments, Formula IV is an amino acid sequence consisting of SEQ ID NO: 51.

[0250] In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula:

[0251] Formula V. R9 - R2 - R6 - R7

[0252] wherein:

[0253] R9 comprises an extracellular domain means for binding ALK;R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4);

[0254] R6 is a protein means derived from toll-like receptor 4 (TLR4); and

[0255] R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0256] In embodiments, R9 of Formula V comprises an extracellular domain means for binding ALK. In embodiments, R9 of Formula V comprises an antibody or antibody fragment. In embodiments, R9 of Formula V comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R9 of Formula V comprises an scFv. In embodiments, R9 of Formula V comprises an scFv that binds ALK. In embodiments, R9 of Formula V comprises SEQ ID NO: 3.

[0257] In embodiments, R9 of Formula V is an scFv. In embodiments, R9 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 3. In embodiments, R9 is an amino acid sequence comprising SEQ ID NO: 3. In embodiments, R9 is an amino acid sequence consisting essentially of SEQ ID NO: 3. In embodiments, R9 is an amino acid sequence consisting of SEQ ID NO: 3.

[0258] In embodiments, R2 of Formula V is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). In embodiments, R2 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 7. In embodiments, R2 is an amino acid sequence comprising SEQ ID NO: 7. In embodiments, R2 is an amino acid sequence consisting essentially of SEQ ID NO: 7. In embodiments, R2 is an amino acid sequence consisting of SEQ ID NO: 7.

[0259] In embodiments, R6 of Formula V is a protein means derived from toll-like receptor 4 (TLR4). In embodiments, R6 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence comprising SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence consisting essentially of SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence consisting of SEQ ID NO: 9.

[0260] In embodiments, R7 of Formula V is an intracellular means derived from toll-like receptor 4 (TLR4). In embodiments, R7 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence comprising SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence consisting essentially of SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence consisting of SEQ ID NO: 11.In embodiments, R9 of Formula V comprises an extracellular domain means for binding ALK of the formula:

[0261] Formula XIV. R24 - R26

[0262] wherein:

[0263] R24 is a protein means derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR); and

[0264] R26 an antibody fragment means for binding ALK.

[0265] In embodiments, R24 of Formula XIV is a protein means derived from granulocytemacrophage colony-stimulating factor receptor (GM-CSFR). In embodiments, R24 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 33. In embodiments, R24 is an amino acid sequence comprising SEQ ID NO: 33. In embodiments, R24 is an amino acid sequence consisting essentially of SEQ ID NO: 33. In embodiments, R24 is an amino acid sequence consisting of SEQ ID NO: 33.

[0266] In embodiments, R26 of Formula XIV an antibody fragment means for binding ALK. In embodiments, R26 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence comprising SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence consisting essentially of SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence consisting of SEQ ID NO: 31.

[0267] In embodiments, the CAR of Formula V is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 57. In embodiments, Formula V is an amino acid sequence comprising SEQ ID NO: 57. In embodiments, Formula V is an amino acid sequence consisting essentially of SEQ ID NO: 57. In embodiments, Formula V is an amino acid sequence consisting of SEQ ID NO: 57.

[0268] In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula:

[0269] Formula VI. R9 - R8 - R6 - R7

[0270] wherein:

[0271] R9 comprises an extracellular domain means for binding ALK;

[0272] R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R6 is a protein means derived from toll-like receptor 4 (TLR4); and

[0273] R7 is an intracellular means derived from toll-like receptor 4 (TLR4).In embodiments, R9 of Formula VI comprises an extracellular domain means for binding ALK. In embodiments, R9 of Formula VI comprises an antibody or antibody fragment. In embodiments, R9 of Formula VI comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R9 of Formula VI comprises an scFv. In embodiments, R9 of Formula VI comprises an scFv that binds ALK. In embodiments, R9 of Formula VI comprises SEQ ID NO: 3.

[0274] In embodiments, R9 of Formula VI is an scFv. In embodiments, R9 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 1. In embodiments, R9 is an amino acid sequence comprising SEQ ID NO: 3. In embodiments, R9 is an amino acid sequence consisting essentially of SEQ ID NO: 3. In embodiments, R9 is an amino acid sequence consisting of SEQ ID NO: 3.

[0275] In embodiments, R8 of Formula VI is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4). In embodiments, R8 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence comprising SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence consisting essentially of SEQ ID NO: 13. In embodiments, R8 is an amino acid sequence consisting of SEQ ID NO: 13.

[0276] In embodiments, R6 of Formula VI is a protein means derived from toll-like receptor 4 (TLR4). In embodiments, R6 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence comprising SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence consisting essentially of SEQ ID NO: 9. In embodiments, R6 is an amino acid sequence consisting of SEQ ID NO: 9.

[0277] In embodiments, R7 of Formula VI is an intracellular means derived from toll-like receptor 4 (TLR4). In embodiments, R7 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence comprising SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence consisting essentially of SEQ ID NO: 11. In embodiments, R7 is an amino acid sequence consisting of SEQ ID NO: 11.

[0278] In embodiments, R9 of Formula VI comprises an extracellular domain means for binding ALK of the formula:

[0279] Formula XIV. R24 - R26

[0280] wherein:R24 is a protein means derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR); and

[0281] R26 an antibody fragment means for binding ALK.

[0282] In embodiments, R24 of Formula XIV is a protein means derived from granulocytemacrophage colony-stimulating factor receptor (GM-CSFR). In embodiments, R24 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 33. In embodiments, R24 is an amino acid sequence comprising SEQ ID NO: 33. In embodiments, R24 is an amino acid sequence consisting essentially of SEQ ID NO: 33. In embodiments, R24 is an amino acid sequence consisting of SEQ ID NO: 33.

[0283] In embodiments, R26 of Formula XIV an antibody fragment means for binding ALK. In embodiments, R26 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence comprising SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence consisting essentially of SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence consisting of SEQ ID NO: 31.

[0284] In embodiments, the CAR of Formula VI is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 59. In embodiments, Formula VI is an amino acid sequence comprising SEQ ID NO: 59. In embodiments, Formula VI is an amino acid sequence consisting essentially of SEQ ID NO: 59. In embodiments, Formula VI is an amino acid sequence consisting of SEQ ID NO: 59.

[0285] In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula:

[0286] Formula VII. R1 -R10 -R11 - R12

[0287] wherein:

[0288] R1 comprises an extracellular binding domain;

[0289] RIO is a hinge or spacer region derived from cluster of differentiation 28 (CD28);

[0290] R11 is a transmembrane domain derived from cluster of differentiation 28 (CD28); and R12 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0291] In embodiments, R1 of Formula VII comprises an extracellular binding domain. In embodiments, R1 of Formula VII comprises an antibody or antibody fragment. In embodiments, R1 of Formula VII comprises an antibody fragment. In embodiments, the antibody fragment comprisesa single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R1 of Formula VII comprises an scFv. In embodiments, R1 of Formula VII comprises an scFv that binds HER2. In embodiments, R1 of Formula VII comprises SEQ ID NO: 1.

[0292] In embodiments, R1 of Formula VII is an scFv. In embodiments, R1 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 1. In embodiments, R1 is an amino acid sequence comprising SEQ ID NO: 1. In embodiments, R1 is an amino acid sequence consisting essentially of SEQ ID NO: 1. In embodiments, R1 is an amino acid sequence consisting of SEQ ID NO: 1.

[0293] In embodiments, RIO of Formula VII is a hinge or spacer region derived from cluster of differentiation 28 (CD28). In embodiments, RIO is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 15. In embodiments, RIO is an amino acid sequence comprising SEQ ID NO: 15. In embodiments, RIO is an amino acid sequence consisting essentially of SEQ ID NO: 15. In embodiments, RIO is an amino acid sequence consisting of SEQ ID NO: 15.

[0294] In embodiments, R11 of Formula VII is a transmembrane domain derived from cluster of differentiation 28 (CD28). In embodiments, R11 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 17. In embodiments, R11 is an amino acid sequence comprising SEQ ID NO: 17. In embodiments, Rll is an amino acid sequence consisting essentially of SEQ ID NO: 17. In embodiments, R11 is an amino acid sequence consisting of SEQ ID NO: 17.

[0295] In embodiments, R12 of Formula VII comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2). In embodiments, R12 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R12 is an amino acid sequence comprising SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R12 is an amino acid sequence consisting essentially of SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R12 is an amino acid sequence consisting of SEQ ID NO: 19 or SEQ ID NO: 21.

[0296] In embodiments, R1 of Formula VII comprises an extracellular binding domain of the formula:

[0297] Formula XI. R20 - R21

[0298] wherein:

[0299] R20 is a signal peptide derived from immunoglobulin K (IgK); and

[0300] R21 is an antibody fragment.In embodiments, R20 of Formula XI is a signal peptide derived from immunoglobulin K (IgK). In embodiments, R20 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence comprising SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence consisting essentially of SEQ ID NO: 27. In embodiments, R20 is an amino acid sequence consisting of SEQ ID NO: 27.

[0301] In embodiments, R21 of Formula XI is an antibody fragment. In embodiments, R21 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 29. In embodiments, R21 is an amino acid sequence comprising SEQ ID NO: 29. In embodiments, R21 is an amino acid sequence consisting essentially of SEQ ID NO: 29. In embodiments, R21 is an amino acid sequence consisting of SEQ ID NO: 29.

[0302] In embodiments, R12 of Formula VII comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) of the formula:

[0303] Formula XVII. R29 - R30

[0304] wherein:

[0305] R29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl); and

[0306] R30 comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2).

[0307] In embodiments, R29 of Formula XVII comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R29 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence comprising SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence consisting essentially of SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence consisting of SEQ ID NO: 37.

[0308] In embodiments, R30 of Formula XVII comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2). In embodiments, R30 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence comprising SEQ ID NO: 39. In embodiments, R30 is an amino acid sequenceconsisting essentially of SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence consisting of SEQ IDNO: 39.

[0309] In embodiments, the linkage between R29 and R30 of Formula XVII is optionally a GS linker. In embodiments, a GS linker has 3 or 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 45).

[0310] In embodiments, R12 of Formula VII comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) of the formula:

[0311] Formula XVIII. R30 - R29

[0312] wherein:

[0313] R30 comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2); and

[0314] R29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl).

[0315] In embodiments, R30 of Formula XVIII comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2). In embodiments, R30 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence comprising SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence consisting essentially of SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence consisting of SEQ IDNO: 39.

[0316] In embodiments, R29 of Formula XVIII comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R29 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence comprising SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence consisting essentially of SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence consisting of SEQ ID NO: 37.

[0317] In embodiments, the linkage between R30 and R29 of Formula XVIII is optionally a GS linker. In embodiments, a GS linker has 3 or 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 3 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGS GGGGS GGGGS (SEQ ID NO: 43).In embodiments, the CAR of Formula VII is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 53 or SEQ ID NO: 55. In embodiments, Formula VII is an amino acid sequence comprising SEQ ID NO: 53 or SEQ ID NO: 55. In embodiments, Formula VII is an amino acid sequence consisting essentially of SEQ ID NO: 53 or SEQ ID NO: 55. In embodiments, Formula VII is an amino acid sequence consisting of SEQ ID NO: 53 or SEQ ID NO: 55.

[0318] In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula:

[0319] Formula VIII. R5 - R13 - R14 - R15

[0320] wherein:

[0321] R5 comprises an extracellular domain means for binding HER2;

[0322] R13 is a protein means derived from cluster of differentiation 28 (CD28) for linking an extracellular means for binding HER2 and a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane;

[0323] R14 is a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane and

[0324] R15 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0325] In embodiments, R5 of Formula VIII comprises an extracellular domain means for binding HER2. In embodiments, R5 of Formula VIII comprises an antibody or antibody fragment. In embodiments, R5 of Formula VIII comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R5 of Formula VIII comprises an scFv. In embodiments, R5 of Formula VIII comprises an scFv that binds HER2. In embodiments, R5 of Formula VIII comprises SEQ ID NO: 1.

[0326] In embodiments, R5 of Formula VIII is an scFv. In embodiments, R5 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence comprising SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence consisting essentially of SEQ ID NO: 1. In embodiments, R5 is an amino acid sequence consisting of SEQ ID NO: 1.In embodiments, R13 of Formula VIII is a protein means derived from cluster of differentiation 28 (CD28) for linking an extracellular means for binding HER2 and a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane. In embodiments, R13 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 15. In embodiments, R13 is an amino acid sequence comprising SEQ ID NO: 15. In embodiments, R13 is an amino acid sequence consisting essentially of SEQ ID NO: 15. In embodiments, R13 is an amino acid sequence consisting of SEQ ID NO: 15.

[0327] In embodiments, R14 of Formula VIII is a protein means derived from cluster of differentiation 28 (CD28) fortraversing a cell membrane . In embodiments, R14 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 17. In embodiments, R14 is an amino acid sequence comprising SEQ ID NO: 17. In embodiments, R14 is an amino acid sequence consisting essentially of SEQ ID NO: 17. In embodiments, R14 is an amino acid sequence consisting of SEQ ID NO: 17.

[0328] In embodiments, R15 of Formula VIII comprises a protein means derived from interferongamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2). In embodiments, R15 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R15 is an amino acid sequence comprising SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R15 is an amino acid sequence consisting essentially of SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R15 is an amino acid sequence consisting of SEQ ID NO: 19 or SEQ ID NO: 21.

[0329] In embodiments, R5 of Formula VIII comprises an extracellular domain means for binding HER.2 of the formula:

[0330] Formula XII. R23 - R25

[0331] wherein:

[0332] R23 is a protein means derived from immunoglobulin K (IgK); and

[0333] R25 an antibody fragment means for binding HER2.

[0334] In embodiments, R23 of Formula XII is a protein means derived from immunoglobulin K (IgK). In embodiments, R23 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence comprising SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence consisting essentially of SEQ ID NO: 27. In embodiments, R23 is an amino acid sequence consisting of SEQ ID NO: 27.In embodiments, R25 of Formula XII an antibody fragment means for binding HER2. In embodiments, R25 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence comprising SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence consisting essentially of SEQ ID NO: 29. In embodiments, R25 is an amino acid sequence consisting of SEQ ID NO: 29.

[0335] In embodiments, R15 of Formula VIII comprises a protein means derived from interferongamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) of the formula:

[0336] Formula XIX. R31 - R32

[0337] wherein:

[0338] R31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl); and R32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2).

[0339] In embodiments, R31 of Formula XIX comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R31 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence comprising SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence consisting essentially of SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence consisting of SEQ ID NO: 37.

[0340] In embodiments, R32 of Formula XIX comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2). In embodiments, R32 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence comprising SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence consisting essentially of SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence consisting of SEQ ID NO: 39.

[0341] In embodiments, the linkage between R31 and R32 of Formula XIX is optionally a GS linker. In embodiments, a GS linker has 3 or 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 45).

[0342] In embodiments, R15 of Formula VIII comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) of the formula:

[0343] Formula XX. R32 - R31

[0344] wherein:R32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2); and R31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl).

[0345] In embodiments, R32 of Formula XX comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2). In embodiments, R32 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence comprising SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence consisting essentially of SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence consisting of SEQ ID NO: 39.

[0346] In embodiments, R31 of Formula XX comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R31 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence comprising SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence consisting essentially of SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence consisting of SEQ ID NO: 37.

[0347] In embodiments, the linkage between R32 and R31 of Formula XX is optionally a GS linker. In embodiments, a GS linker has 3 or 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 3 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGS GGGGS GGGGS (SEQ ID NO: 43).

[0348] In embodiments, the CAR of Formula VIII is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 53 or SEQ ID NO: 55. In embodiments, Formula VIII is an amino acid sequence comprising SEQ ID NO: 53 or SEQ ID NO: 55. In embodiments, Formula VIII is an amino acid sequence consisting essentially of SEQ ID NO: 53 or SEQ ID NO: 55. In embodiments, Formula VIII is an amino acid sequence consisting of SEQ ID NO: 53 or SEQ ID NO: 55.

[0349] In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula:

[0350] Formula IX. R1 - R16 - R17 - R12

[0351] wherein:

[0352] R1 comprises an extracellular binding domain;

[0353] R16 is a hinge or spacer region derived from cluster of differentiation 8 (CD8);

[0354] R17 is a transmembrane domain derived from interferon-gamma receptor 1 (IFNgRl); andR12 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0355] In embodiments, R1 of Formula IX comprises an extracellular binding domain. In embodiments, R1 of Formula IX comprises an antibody or antibody fragment. In embodiments, R1 of Formula IX comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R1 of Formula IX comprises an scFv. In embodiments, R1 of Formula IX comprises an scFv that binds ALK. In embodiments, R1 of Formula IX comprises SEQ ID NO: 5.

[0356] In embodiments, R1 of Formula IX is an scFv. In embodiments, R1 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 1. In embodiments, R1 is an amino acid sequence comprising SEQ ID NO: 5. In embodiments, R1 is an amino acid sequence consisting essentially of SEQ ID NO: 5. In embodiments, R1 is an amino acid sequence consisting of SEQ ID NO: 5.

[0357] In embodiments, R16 of Formula IX is a hinge or spacer region derived from cluster of differentiation 8 (CD8). In embodiments, R16 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 23. In embodiments, R16 is an amino acid sequence comprising SEQ ID NO: 23. In embodiments, R16 is an amino acid sequence consisting essentially of SEQ ID NO: 23. In embodiments, R16 is an amino acid sequence consisting of SEQ ID NO: 23.

[0358] In embodiments, R17 of Formula IX is a transmembrane domain derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R17 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 25. In embodiments, R17 is an amino acid sequence comprising SEQ ID NO: 25. In embodiments, R17 is an amino acid sequence consisting essentially of SEQ ID NO: 25. In embodiments, R17 is an amino acid sequence consisting of SEQ ID NO: 25.

[0359] In embodiments, R12 of Formula IX comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2). In embodiments, R12 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R12 is an amino acid sequence comprising SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R12 is an amino acid sequence consistingessentially of SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R12 is an amino acid sequence consisting of SEQ ID NO: 19 or SEQ ID NO: 21.

[0360] In embodiments, R1 of Formula IX comprises an extracellular binding domain of the formula:

[0361] Formula XV. R27 - R21

[0362] wherein:

[0363] R27 is a signal peptide derived from cluster of differentiation 8 (CD8); and

[0364] R21 is an antibody fragment.

[0365] In embodiments, R27 of Formula XV is a signal peptide derived from cluster of differentiation 8 (CD8). In embodiments, R27 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 35. In embodiments, R27 is an amino acid sequence comprising SEQ ID NO: 35. In embodiments, R27 is an amino acid sequence consisting essentially of SEQ ID NO: 35. In embodiments, R27 is an amino acid sequence consisting of SEQ ID NO: 35.

[0366] In embodiments, R21 of Formula XI is an antibody fragment. In embodiments, R21 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence comprising SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence consisting essentially of SEQ ID NO: 31. In embodiments, R21 is an amino acid sequence consisting of SEQ ID NO: 31.

[0367] In embodiments, R12 of Formula IX comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) of the formula:

[0368] Formula XVII. R29 - R30

[0369] wherein:

[0370] R29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl); and

[0371] R30 comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2).

[0372] In embodiments, R29 of Formula XVII comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R29 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 37. In embodiments, R29 is anamino acid sequence comprising SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence consisting essentially of SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence consisting of SEQ ID NO: 37.

[0373] In embodiments, R30 of Formula XVII comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2). In embodiments, R30 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence comprising SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence consisting essentially of SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence consisting of SEQ ID NO: 39.

[0374] In embodiments, the linkage between R29 and R30 of Formula XVII is optionally a GS linker. In embodiments, a GS linker has 3 or 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGS GGGGS GGGGS GGGGS (SEQ ID NO: 45).

[0375] In embodiments, R12 of Formula IX comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) of the formula:

[0376] Formula XVIII. R30 - R29

[0377] wherein:

[0378] R30 comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2); and

[0379] R29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl).

[0380] In embodiments, R30 of Formula XVIII comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2). In embodiments, R30 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence comprising SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence consisting essentially of SEQ ID NO: 39. In embodiments, R30 is an amino acid sequence consisting of SEQ ID NO: 39.

[0381] In embodiments, R29 of Formula XVIII comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R29 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence comprising SEQ ID NO: 37. In embodiments, R29 is an amino acid sequenceconsisting essentially of SEQ ID NO: 37. In embodiments, R29 is an amino acid sequence consisting of SEQ IDNO: 37.

[0382] In embodiments, the linkage between R30 and R29 of Formula XVIII is optionally a GS linker. In embodiments, a GS linker has 3 or 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 3 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGS GGGGS GGGGS (SEQ ID NO: 43).

[0383] In embodiments, the CAR of Formula IX is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 61 or SEQ ID NO: 63. In embodiments, Formula IX is an amino acid sequence comprising SEQ ID NO: 61 or SEQ ID NO: 63. In embodiments, Formula IX is an amino acid sequence consisting essentially of SEQ ID NO: 61 or SEQ ID NO: 63. In embodiments, Formula IX is an amino acid sequence consisting of SEQ ID NO: 61 or SEQ ID NO: 63.

[0384] In embodiments, the CAR is of the formula, or the nucleic acids comprise a nucleic acid sequence encoding a CAR of the formula:

[0385] Formula X . R9 - R18 - R19 - R15

[0386] wherein:

[0387] R9 comprises an extracellular domain means for binding ALK;

[0388] R18 is a protein means derived from cluster of differentiation 8 (CD8);

[0389] R19 is a protein means derived from interferon-gamma receptor 1 (IFNgRl); and

[0390] R15 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0391] In embodiments, R9 of Formula X comprises an extracellular domain means for binding ALK. In embodiments, R9 of Formula X comprises an antibody or antibody fragment. In embodiments, R9 of Formula X comprises an antibody fragment. In embodiments, the antibody fragment comprises a single-chain variable region fragment (scFv) or a single heavy chain variable region fragment (also known as a nanobody or VHH). In embodiments, R9 of Formula X comprises an scFv. In embodiments, R9 of Formula X comprises an scFv that binds ALK. In embodiments, R9 of Formula X comprises SEQ ID NO: 1.

[0392] In embodiments, R9 of Formula X is an scFv. In embodiments, R9 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 5. In embodiments, R9 is an amino acid sequence comprising SEQ ID NO: 5. In embodiments, R9 is an amino acid sequenceconsisting essentially of SEQ ID NO: 5. In embodiments, R9 is an amino acid sequence consisting of SEQ IDNO: 5.

[0393] In embodiments, R18 of Formula X is a protein means derived from cluster of differentiation 8 (CD8). In embodiments, R18 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 23. In embodiments, R18 is an amino acid sequence comprising SEQ ID NO: 23. In embodiments, R18 is an amino acid sequence consisting essentially of SEQ ID NO: 23. In embodiments, R18 is an amino acid sequence consisting of SEQ ID NO: 23.

[0394] In embodiments, R19 of Formula X is a protein means derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R19 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 25. In embodiments, R19 is an amino acid sequence comprising SEQ ID NO: 25. In embodiments, R19 is an amino acid sequence consisting essentially of SEQ ID NO: 25. In embodiments, R19 is an amino acid sequence consisting of SEQ ID NO: 25.

[0395] In embodiments, R15 of Formula X comprises a protein means derived from interferongamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2). In embodiments, R15 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R15 is an amino acid sequence comprising SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R15 is an amino acid sequence consisting essentially of SEQ ID NO: 19 or SEQ ID NO: 21. In embodiments, R15 is an amino acid sequence consisting of SEQ ID NO: 19 or SEQ ID NO: 21.

[0396] In embodiments, R9 of Formula X comprises an extracellular domain means for binding ALK of the formula:

[0397] Formula XVI. R28 - R26

[0398] wherein:

[0399] R28 is a protein means derived from cluster of differentiation 8 (CD8); and

[0400] R26 an antibody fragment means for binding ALK.

[0401] In embodiments, R28 of Formula XVI is a protein means derived from cluster of differentiation 8 (CD8). In embodiments, R28 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 35. In embodiments, R28 is an amino acid sequence comprising SEQ ID NO: 35. In embodiments, R28 is an amino acid sequence consisting essentially of SEQ ID NO: 35. In embodiments, R28 is an amino acid sequence consisting of SEQ ID NO: 35.In embodiments, R26 of Formula XVI an antibody fragment means for binding ALK. In embodiments, R26 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence comprising SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence consisting essentially of SEQ ID NO: 31. In embodiments, R26 is an amino acid sequence consisting of SEQ ID NO: 31.

[0402] In embodiments, R15 of Formula X comprises a protein means derived from interferongamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) of the formula:

[0403] Formula XIX. R31 - R32

[0404] wherein:

[0405] R31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl); and R32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2).

[0406] In embodiments, R31 of Formula XIX comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R31 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence comprising SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence consisting essentially of SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence consisting of SEQ ID NO: 37.

[0407] In embodiments, R32 of Formula XIX comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2). In embodiments, R32 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence comprising SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence consisting essentially of SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence consisting of SEQ ID NO: 39.

[0408] In embodiments, the linkage between R31 and R32 of Formula XIX is optionally a GS linker. In embodiments, a GS linker has 3 or 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 45).

[0409] In embodiments, R15 of Formula X comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2) of the formula:

[0410] Formula XX. R32 - R31

[0411] wherein:R32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2); and R31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl).

[0412] In embodiments, R32 of Formula XIX comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2). In embodiments, R32 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence comprising SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence consisting essentially of SEQ ID NO: 39. In embodiments, R32 is an amino acid sequence consisting of SEQ ID NO: 39.

[0413] In embodiments, R31 of Formula XIX comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl). In embodiments, R31 is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence comprising SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence consisting essentially of SEQ ID NO: 37. In embodiments, R31 is an amino acid sequence consisting of SEQ ID NO: 37.

[0414] In embodiments, the linkage between R32 and R31 of Formula XX is optionally a GS linker. In embodiments, a GS linker has 3 or 4 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker has 3 repeats of the sequence GGGGS (SEQ ID NO: 41). In embodiments, a GS linker is GGGGSGGGGSGGGGS (SEQ ID NO: 43).

[0415] In embodiments, the CAR of Formula X is an amino acid sequence comprising, consisting essentially of, or consisting of SEQ ID NO: 61 or SEQ ID NO: 63. In embodiments, Formula X is an amino acid sequence comprising SEQ ID NO: 61 or SEQ ID NO: 63. In embodiments, Formula X is an amino acid sequence consisting essentially of SEQ ID NO: 61 or SEQ ID NO: 63. In embodiments, Formula X is an amino acid sequence consisting of SEQ ID NO: 61 or SEQ ID NO: 63.

[0416] In embodiments, chimeric antigen receptors provided herein are encoded by a vector, such as a vector comprising a nucleic acid or nucleic acids provided herein. In embodiments, chimeric antigen receptors or vectors comprising the same provided herein are encoded with codon-optimized nucleic acids. In embodiments, chimeric antigen receptors provided herein are encoded by a plasmid vector, such as a plasmid vector comprising a nucleic acid or nucleic acids provided herein. In embodiments, chimeric antigen receptors provided herein are encoded by a lentiviral plasmid vector,such as a lentiviral plasmid vector comprising a nucleic acid or nucleic acids provided herein. Other exemplary vectors are understood in the art, such as adeno-associated virus vectors.

[0417] Provided herein are engineered macrophages and pharmaceutical compositions thereof. Engineered macrophages and pharmaceutical compositions are made in a variety of ways understood by a person of ordinary skill in the art. In embodiments, a vector (e.g., a vector comprising a nucleic acid or nucleic acids provided herein) is transduced into a macrophage ex vivo, such as by a viral or retroviral vector (e.g., a viral or retroviral vector comprising the nucleic acids provided herein). In embodiments, a lentiviral vector (e.g., a lentiviral vector comprising a nucleic acid or nucleic acids provided herein) is transduced ex vivo directly into a primary monocyte or macrophage (see, e.g., U.S. Pat. App. Pub. No. 2024 / 0066126, incorporated herein by reference in its entirety on March 25, 2025; Sloas et al. (2021) Engineered CAR Macrophages as Adoptive Immunotherapies for Solid Tumors. Front. Immunol. 12:783305. doi: 10.3389 / fimmu.2021.783305; Zhang et al., Engineering CAR-T cells, Biomark Res. 5:22 (2017)). Other methods of engineering macrophages are understood in the art, such as in vivo methods of engineering (see, e.g., U.S. Pat. App. Pub. No. 2024 / 0066126, incorporated herein by reference in its entirety on March 25, 2025; Sloas et al. (2021) Engineered CAR Macrophages as Adoptive Immunotherapies for Solid Tumors. Front. Immunol. 12:783305. doi: 10.3389 / fimmu.2021.783305; Dabiri et al., Site-specific transgene integration in chimeric antigen receptor (CAR) T cell therapies, Biomarker Research 11 : 67 (2023)). In embodiments, pharmaceutical compositions provided herein comprise engineered macrophages provided herein and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0418] Provided herein are engineered macrophages and pharmaceutical compositions. A person of ordinary skill in the art understands that engineered macrophages or pharmaceutical compositions provided herein can be formulated and administered in a variety of ways. In embodiments, engineered macrophages and pharmaceutical compositions provided herein are formulated for administration by any suitable route, such as intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, subcutaneous, intratumoral, epidural, nasal, oral, vaginal, rectal, topical, local, systemic, otic, inhalational, buccal (e.g., sublingual), and transdermal and can include additional biologically active agents, can facilitate or enhance delivery, or can control release. Other biologically active agents can also be administered sequentially, intermittently or simultaneously, such as in the same composition. Controlled release formulations and devices, such as by pump, are contemplated. Additional means of formulation, administration, storage, preparation, and manufacturing are contemplated (e.g., U.S. Pat. App. Pub. No. 2024 / 0066126, incorporated hereinby reference in its entirety on March 25, 2025; U.S. Pat. App. Pub. No. 2020 / 0215123, incorporated herein by reference in its entirety on March 25, 2025).

[0419] In embodiments, engineered macrophages or pharmaceutical compositions provided herein can be formulated into suitable pharmaceutical preparations, such as solutions, suspensions, tablets, dispersible tablets, pills, capsules, powders, patches, or sustained release formulations. In embodiments, engineered macrophages or pharmaceutical compositions provided herein are formulated in a dried or liquid form. In embodiments, engineered cells or pharmaceutical compositions provided herein are formulated in a liquid form, for example, as a suspension for injection (direct administration) or frozen suspension that is thawed prior to use, dried soluble form, and emulsion. In embodiments, injection or infusion administration is contemplated; for example, subcutaneous, intramuscular, intratumoral, intravenous, or intradermal administration is contemplated. In embodiments, excipients, such as water, saline, dextrose, or glycerol, and carriers, such as diluents, adjuvants, anti-adherents, binders, coatings, fillers, flavors, colors, lubricants, glidants, preservatives, detergents, sorbents or combinations thereof, are contemplated. In embodiments, aqueous vehicles, non-aqueous vehicles, isotonic agents, buffers, antioxidants, local anesthetics, suspending agents, dispersing agents, emulsifying agents, sequestering agents, chelating agents, or combinations thereof are contemplated herein as excipients and carriers. In embodiments, a therapeutically effective amount of engineered macrophages or pharmaceutical compositions provided herein are formulated, for example, as single-unit or multi-unit dosage formulations.

[0420] Additional formulations and modes of administration are contemplated (e.g., U.S. Pat. App. Pub. No.

[0421] 2024 / 0066126, incorporated herein by reference in its entirety on March 25, 2025; U.S. Pat. App. Pub. No. 2020 / 0215123, incorporated herein by reference in its entirety on March 25, 2025).

[0422] Methods

[0423] Provided herein are engineered macrophages and pharmaceutical compositions comprising the engineered macrophages disclosed herein and one or more acceptable carriers, diluents, or excipients. Further provided herein are methods of treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, nonsmall cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma,rhabdomyosarcoma, renal cell carcinoma) a pharmaceutical composition provided herein. Further provided herein are methods of treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein. Particularly, provided herein are methods of treating cancer selected from the list consisting of: breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of a pharmaceutical composition comprising engineered macrophage provided herein. Particularly, the provided herein are methods of treating cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of: breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma,rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein.

[0424] In embodiments, provided herein are methods of treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer, such as neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) a pharmaceutical composition provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Further provided herein are methods of treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer, such as neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, provided herein are methods of treating cancer selected from the list consisting of: neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of a pharmaceutical composition comprising engineered macrophage provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, the provided herein are methods of treating cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer,squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of: neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31).

[0425] In embodiments, provided herein are methods of treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) a pharmaceutical composition provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Further provided herein are methods of treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer, such as breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein are methods of treating cancer selected from the list consisting of: breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of a pharmaceutical composition comprising engineered macrophage provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, the provided herein are methods of treating cancer selected from the list consisting of breast cancer, bladdercancer, pancreatic cancer, ovarian cancer, and stomach cancer comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of: breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29).

[0426] Further provided herein are pharmaceutical compositions provided herein for use in treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) a pharmaceutical composition provided herein. Further provided herein are engineered macrophages provided herein for use in treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein. Particularly, provided herein are pharmaceutical compositions provided herein for use in treating cancer selected from the list consisting breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large celllymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of a pharmaceutical composition comprising engineered macrophage provided herein. Particularly, provided herein are pharmaceutical compositions provided herein for use in treating cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein.

[0427] Further provided herein are pharmaceutical compositions provided herein for use in treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) a pharmaceutical composition provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e g., SEQ ID NO: 31). Further provided herein are engineered macrophages provided herein for use in treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, provided herein are pharmaceutical compositions provided herein for use in treatingcancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of a pharmaceutical composition comprising engineered macrophage provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, provided herein are pharmaceutical compositions provided herein for use in treating cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31).

[0428] Further provided herein are pharmaceutical compositions provided herein for use in treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) a pharmaceutical composition provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HERZ or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Further provided herein are engineered macrophages provided herein for use in treating cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladdercancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein are pharmaceutical compositions provided herein for use in treating cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of a pharmaceutical composition comprising engineered macrophage provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein are pharmaceutical compositions provided herein for use in treating cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29).

[0429] Further provided herein are uses of a pharmaceutical composition provided herein for the treatment of cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) a pharmaceutical composition provided herein. Further provided herein are uses of an engineered macrophageprovided herein for the treatment of cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein. Particularly, provided herein are uses of a pharmaceutical composition provided herein for the treatment of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of a pharmaceutical composition comprising an engineered macrophage provided herein. Particularly, provided herein are uses of an engineered macrophage provided herein for the treatment of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma,rhabdomyosarcoma, renal cell carcinoma) an effective amount of an engineered macrophage provided herein.

[0430] Further provided herein are uses of a pharmaceutical composition provided herein for the treatment of cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) a pharmaceutical composition provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Further provided herein are uses of an engineered macrophage provided herein for the treatment of cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, provided herein are uses of a pharmaceutical composition provided herein for the treatment of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of a pharmaceutical composition comprising an engineered macrophage provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31).

[0431] Particularly, provided herein are uses of an engineered macrophage provided herein for the treatment of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplasticlarge cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of an engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31).

[0432] Further provided herein are uses of a pharmaceutical composition provided herein for the treatment of cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) a pharmaceutical composition provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Further provided herein are uses of an engineered macrophage provided herein for the treatment of cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein are uses of a pharmaceutical composition provided herein for the treatment of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of a pharmaceutical composition comprising an engineered macrophage provided herein,such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein are uses of an engineered macrophage provided herein for the treatment of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of an engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29).

[0433] Further provided herein are pharmaceutical compositions for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) a pharmaceutical composition provided herein. Further provided herein are engineered macrophages for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein. Particularly, provided herein pharmaceutical compositions for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiatedanaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of a pharmaceutical composition comprising an engineered macrophage provided herein. Particularly, provided herein are engineered macrophages for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein.

[0434] Further provided herein are pharmaceutical compositions for use in therapy, comprising administering to a patient or subject in need thereof (e g., a patient or subject with or at risk of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) a pharmaceutical composition provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Further provided herein are engineered macrophages for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, provided herein pharmaceutical compositions for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of a pharmaceutical composition comprising an engineered macrophage provided herein, such as a pharmaceutical composition comprising anengineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, provided herein are engineered macrophages for use in therapy, comprising administering to a patient or subject in need thereof (eg., a patient or subject with or at risk of cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31).

[0435] Further provided herein are pharmaceutical compositions for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) a pharmaceutical composition provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Further provided herein are engineered macrophages for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein pharmaceutical compositions for use in therapy, comprising administering to a patient or subject in need thereof (e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of a pharmaceutical composition comprising an engineered macrophage provided herein, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein are engineered macrophages for use in therapy, comprising administering to a patient or subject in need thereof(e.g., a patient or subject with or at risk of cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer) an effective amount of engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29).

[0436] Further provided herein are uses of pharmaceutical compositions provided herein in the manufacture of a medicament for treating cancer (e.g., a cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma). Further provided herein are uses of engineered macrophages provided herein in the manufacture of a medicament for treating cancer (e.g., a cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma). Particularly, provided herein are uses of pharmaceutical compositions provided herein in the manufacture of a medicament for treating cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma. Particularly, provided herein are uses of engineered macrophages provided herein in the manufacture of a medicament for treating cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma.Further provided herein are uses of pharmaceutical compositions provided herein in the manufacture of a medicament for in treating cancer (e.g., a cancer selected from the list consisting neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma), such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Further provided herein are uses of engineered macrophages provided herein in the manufacture of a medicament for in treating cancer (e.g., a cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma), such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, provided herein are uses of pharmaceutical compositions provided herein in the manufacture of a medicament for in treating cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Particularly, provided herein are uses of engineered macrophages provided herein in the manufacture of a medicament for in treating cancer selected from the list consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31).

[0437] Further provided herein are uses of pharmaceutical compositions provided herein in the manufacture of a medicament for in treating cancer (e.g., a cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervicalcancer), such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Further provided herein are uses of engineered macrophages provided herein in the manufacture of a medicament for in treating cancer (e g., a cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer), such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein are uses of pharmaceutical compositions provided herein in the manufacture of a medicament for in treating cancer selected from the list consisting breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, such as a pharmaceutical composition comprising an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Particularly, provided herein are uses of engineered macrophages provided herein in the manufacture of a medicament for in treating cancer selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29).

[0438] Further provided herein are pharmaceutical compositions for use in the treatment of cancer (e.g., breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma), comprising an engineered macrophage provided herein. Further provided herein are pharmaceutical compositions for use in the treatment of cancer selected from the listed consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, neuroblastoma, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiatedanaplastic thyroid carcinoma, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising an engineered macrophage provided herein.

[0439] Further provided herein are pharmaceutical compositions for use in the treatment of cancer (e.g., neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma), comprising an engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31). Further provided herein are pharmaceutical compositions for use in the treatment of cancer selected from the listed consisting of neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma, comprising an engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds ALK or an scFv means for binding ALK (e.g., SEQ ID NO: 31).

[0440] Further provided herein are pharmaceutical compositions for use in the treatment of cancer (e.g., breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer), comprising an engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29). Further provided herein are pharmaceutical compositions for use in the treatment of cancer selected from the listed consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer, comprising an engineered macrophage provided herein, such as an engineered macrophage comprising a nucleic acid encoding an scFv that binds HER2 or an scFv means for binding HER2 (e.g., SEQ ID NO: 29).

[0441] EXAMPLES

[0442] The EXAMPLES show embodiments that include chimeric antigen receptor (CAR) variations as disclosed herein. The reporter cell line in the EXAMPLES is used to detect CAR-driven activation of various signaling pathways, e.g., nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB), interferon regulatory factor 1 (IRF1), and nuclear factor of activated T-cells (NFAT) pathways (FIGS. 1, 2A-2C, and 3A).First, a human myelomonocytic cell line was established. The cell line was named U937 and contained fluorescent reporters for the activation of key signaling pathways, e.g., nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB), interferon regulatory factor 1 fIR.FI), and nuclear factor of activated T-cells (NF AT), which are central in shaping the macrophages’ immune response (FIGS. 1, 2A-2C, and 3 A). The functionality and the dynamic range of the reporter system was assessed by flow cytometry after activation with specific synthetic stimuli, including ionomycin (NF AT activation), interferon-gamma (IFNg, IRF1 activation) and lipopolysaccharide (LPS, NFkB activation)(FIG.l). The level of activation was expressed as “fold change” (FC) in mean fluorescence intensity (MFI) over unstimulated control.

[0443] U937 reporter cells were transduced with lentiviral vectors encoding ALK-CARs with the different intracellular signaling domain configurations. Transduced U937c cells were mixed with ALK-expressing target cells as a specific CAR-stimulant or with ALK -negative cells as control to assess background activation in the absence of specific CAR activation (tonic signaling) (FIGs. 2A, 2B, and 2C). CAR-TLR4 variants were identified as potent activators of the NFkB pathway, although the activation did not depend on specific activation by ALK (FIG. 2B). Two CAR variants containing IFNg derived signaling domains also showed moderate, but specific activation of the NFkB or NF AT pathways. To address the tonic signaling by the TLR4-intracellular signaling domain (ICD), a variant including a portion of the extracellular domain (aa545TLR4) was tested. This variant achieved strong and antigen-dependent activation of the NFkB pathway (FIGs. 3A and 3B).

[0444] Next, human CD34+ hematopoietic stem cells were transduced with lentiviral vectors encoding the ALK-CAR variants, followed by in vitro differentiation into macrophages with G-CSF and GM-CSF. Mature CAR-macrophages were incubated with NB1 ALK-expressing neuroblastoma cells for CAR-stimulation. The physiological consequences on macrophage polarization was assessed via staining for surface markers indicative of pro- and anti-inflammatory phenotypes and on antigen-presentation. Consistent upregulation of markers associated with antigen presentation was observed across tested variants (FIG. 4). Some IFNg-ICD variants additionally showed strong upregulation of inflammatory Ml-type associated markers. A subset of variants was tested in the context of a CAR targeting the cancer antigen HER2. Stimulation with HER2 -positive target cells again resulted in strong activation of antigen presentation associated markers and IFNg-signaling showed induction of an inflammatory phenotype (FIG. 5).Next, the ability of CAR-mac to phagocytose and eliminate tumor cells was tested. Human CD34+ hematopoietic stem cells were transduced with lentiviral vectors encoding the ALK-CAR or HER2-CAR variants, followed by in vitro differentiation into macrophages with G-CSF and GM-CSF. Mature CAR-macrophages were incubated with fluorescent tumor cells expressing the ALK or HERZ antigen, respectively. Phagocytic uptake of NB cells was assessed by flow cytometry. Very high phagocytosis rates were observed for HER2-d542 and d545 variants, and to a lesser extent also for IFNg-ICD variants IFNg9 and 10 (FIG. 6).

[0445] In sum, the EXAMPLES show assays demonstrating CAR functionality in primary macrophages by co-culturing a neuroblastoma cell line with dNGFR-PGK-aALKCAR-ICD lentiviral transfected human CD34 progenitor (hCD34+) cells. CAR-mediated polarization based on surface marker expression is then assayed in CARs targeting tumor antigens anaplastic lymphoma kinase (ALK) (FIGS. 4 and 6) and human epidermal growth factor receptor 2 (HER2) (FIGS. 5 and 6).

[0446] EMBODIMENTS

[0447] Embodiment l is a macrophage chimeric antigen receptor (CAR) of the formula:

[0448] Formula I. R1 - R2 - R3 - R4

[0449] wherein: R1 comprises an extracellular binding domain; R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R3 is a transmembrane domain derived from tolllike receptor 4 (TLR4); and R4 comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4).

[0450] Embodiment 2 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0451] Formula II. R5 - R2 - R6 - R7

[0452] wherein: R5 comprises an extracellular domain means for binding HER2; R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R6 is a protein means derived from toll-like receptor 4 (TLR4); and R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0453] Embodiment 3 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0454] Formula III. R1 - R8 - R3 - R4

[0455] wherein: R1 comprises an extracellular binding domain; R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R3 is a transmembrane domain derived from toll-like receptor 4 (TLR4); and R4 comprises an intracellular signaling domain derived from tolllike receptor 4 (TLR4).Embodiment 4 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0456] Formula IV. R5 - R8 - R6 - R7

[0457] wherein: R5 comprises an extracellular domain means for binding HER2; R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R6 is a protein means derived from toll-like receptor 4 (TLR4); and R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0458] Embodiment 5 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0459] Formula V. R9 - R2 - R6 - R7

[0460] wherein: R9 comprises an extracellular domain means for binding ALK; R2 is a truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R6 is a protein means derived from toll-like receptor 4 (TLR4); and R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0461] Embodiment 6 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0462] Formula VI. R9 - R8 - R6 - R7

[0463] wherein: R9 comprises an extracellular domain means for binding ALK; R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R6 is a protein means derived from toll-like receptor 4 (TLR4); and R7 is an intracellular means derived from toll-like receptor 4 (TLR4).

[0464] Embodiment 7 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0465] Formula VII. R1 - RIO - R11 - R12

[0466] wherein: R1 comprises an extracellular binding domain; RIO is a hinge or spacer region derived from cluster of differentiation 28 (CD28); R11 is a transmembrane domain derived from cluster of differentiation 28 (CD28); and R12 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0467] Embodiment 8 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0468] Formula VIII. R5 - R13 - R14 - R15

[0469] wherein: R5 comprises an extracellular domain means for binding HER2; R13 is a protein means derived from cluster of differentiation 28 (CD28) for linking an extracellular means for binding HER2 and a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane; R14 is a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane and R15 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).Embodiment 9 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0470] Formula IX. R1 - R16 - R17 - R12

[0471] wherein: R1 comprises an extracellular binding domain; R16 is a hinge or spacer region derived from cluster of differentiation 8 (CD8); R17 is a transmembrane domain derived from interferon-gamma receptor 1 (IFNgRl); and R12 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0472] Embodiment 10 is a macrophage chimeric antigen receptor (CAR) of the formula:

[0473] Formula X. R9 - R18 - R19 - R15

[0474] wherein: R9 comprises an extracellular domain means for binding ALK; R18 is a protein means derived from cluster of differentiation 8 (CD8); R19 is a protein means derived from interferon-gamma receptor 1 (IFNgRl ); and R15 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2).

[0475] Embodiment 11 is the macrophage CAR of any one of embodiments 1, 3, or 7, wherein R1 is of the formula:

[0476] Formula XI R20 - R21

[0477] wherein: R20 is a signal peptide derived from immunoglobulin K (IgK); and R21 is an antibody fragment.

[0478] Embodiment 12 is the macrophage CAR of embodiment 1 or embodiment 3, wherein R1 is of the formula:

[0479] Formula XIII. R22 - R21

[0480] wherein: R22 is a signal peptide derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR); and R21 is an antibody fragment.

[0481] Embodiment 13 is the macrophage CAR of any one of embodiments 2, 4, or 8, wherein R5 is of the formula:

[0482] Formula XII. R23 - R25

[0483] wherein: R23 is a protein means derived from immunoglobulin K (IgK); and R25 an antibody fragment means for binding HER2.

[0484] Embodiment 14 is the macrophage CAR of embodiment 5 or embodiment 6, wherein R9 is of the formula:

[0485] Formula XIV. R24 - R26wherein: R24 is a protein means derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR); and R26 an antibody fragment means for binding ALK.

[0486] Embodiment 15 is the macrophage CAR of embodiment 7 or embodiment 9, wherein R12 is of the formula:

[0487] Formula XVII. R29 - R30

[0488] wherein: R29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl); and R30 comprises an intracellular signaling domain derived from interferongamma receptor 2 (IFNgR2).

[0489] Embodiment 16 is the macrophage CAR of embodiment 7 or embodiment 9, wherein R12 is of the formula:

[0490] Formula XVIII. R30 - R29

[0491] wherein: R30 comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2); and R29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl).

[0492] Embodiment 17 is the macrophage CAR of embodiment 8 or embodiment 10, wherein R15 is of the formula:

[0493] Formula XIX. R31 - R32

[0494] wherein: R31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl); and R32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2).

[0495] Embodiment 18 is the macrophage CAR of embodiment 8 or embodiment 10, wherein R15 is of the formula:

[0496] Formula XX. R32 - R31

[0497] wherein: R32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2); and R31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl).

[0498] Embodiment 19 is the macrophage CAR of embodiment 9, wherein R1 is of the formula: Formula XV R27 - R21

[0499] wherein: R27 is a signal peptide derived from cluster of differentiation 8 (CD8); and R21 is an antibody fragment.

[0500] Embodiment 20 is the macrophage CAR of embodiment 10, wherein R9 is of the formula: Formula XVI. R28 - R26

[0501] wherein: R28 is a protein means derived from cluster of differentiation 8 (CD8); and R26 an antibody fragment means for binding ALK.Embodiment 21 is the macrophage CAR of any one of embodiments 1, 3, or 7, wherein R1 comprises, consists essentially of, or consists of SEQ ID NO: 1.

[0502] Embodiment 22 is the macrophage CAR of any one of embodiments 2, 4, or 8, wherein R5 comprises, consists essentially of, or consists of SEQ ID NO: 1.

[0503] Embodiment 23 is the macrophage CAR of embodiment 1 or embodiment 3, wherein R1 comprises, consists essentially of, or consists of SEQ ID NO: 3.

[0504] Embodiment 24 is the macrophage CAR of embodiment 9, wherein R1 comprises, consists essentially of, or consists of SEQ ID NO: 5

[0505] Embodiment 25 is the macrophage CAR of embodiment 5 or embodiment 6, wherein R9 comprises, consists essentially of, or consists of SEQ ID NO: 3.

[0506] Embodiment 26 is the macrophage CAR of embodiment 10, wherein R9 comprises, consists essentially of, or consists of SEQ ID NO: 5.

[0507] Embodiment 27 is the macrophage CAR of embodiment 11, wherein R21 comprises, consists essentially of, or consists of SEQ ID NO: 29.

[0508] Embodiment 28 is the macrophage CAR of embodiment 12, wherein R21 comprises, consists essentially of, or consists of SEQ ID NO: 31.

[0509] Embodiment 29 is the macrophage CAR of embodiment 13, wherein R25 comprises, consists essentially of, or consists of SEQ ID NO: 29.

[0510] Embodiment 30 is the macrophage CAR of embodiment 14, wherein R26 comprises, consists essentially of, or consists of SEQ ID NO: 31.

[0511] Embodiment 31 is the macrophage CAR of embodiment 14, wherein R26 comprises, consists essentially of, or consists of SEQ ID NO: 31.

[0512] Embodiment 32 is the macrophage CAR of embodiment 7 or embodiment 9, wherein R12 comprises, consists essentially of, or consists of SEQ ID NO: 19 or SEQ ID NO: 21.

[0513] Embodiment 33 is the macrophage CAR of embodiment 8 or embodiment 10, wherein R15 comprises, consists essentially of, or consists of SEQ ID NO: 19 or SEQ ID NO: 21.

[0514] Embodiment 34 is the macrophage CAR of any one of embodiments 1-2, and 5, wherein R2 comprises, consists essentially of, or consists of SEQ ID NO: 7.

[0515] Embodiment 35 is the macrophage CAR of embodiment 1 or embodiment 3, wherein R3 comprises, consists essentially of, or consists of SEQ ID NO: 9.

[0516] Embodiment 36 is the macrophage CAR of embodiment 1 or embodiment 3, wherein R4 comprises, consists essentially of, or consists of SEQ ID NO: 11.Embodiment 37 is the CAR of any one of embodiments 2 and 4-6, wherein R6 comprises, consists essentially of, or consists of SEQ ID NO: 9.

[0517] Embodiment 38 is the macrophage CAR of any one of embodiments 2 and 4-6, wherein R7 comprises, consists essentially of, or consists of SEQ ID NO: 11.

[0518] Embodiment 39 is the macrophage CAR of any one of embodiments 3, 4, and 6, wherein R8 comprises, consists essentially of, or consists of SEQ ID NO: 13.

[0519] Embodiment 40 is the macrophage CAR of embodiment 7, wherein RIO comprises, consists essentially of, or consists of SEQ ID NO: 15.

[0520] Embodiment 41 is the macrophage CAR of embodiment 7, wherein R11 comprises, consists essentially of, or consists of SEQ ID NO: 17.

[0521] Embodiment 42 is the macrophage CAR of embodiment 8, wherein R13 comprises, consists essentially of, or consists of SEQ ID NO: 15.

[0522] Embodiment 43 is the macrophage CAR of embodiment 8, wherein R14 comprises, consists essentially of, or consists of SEQ ID NO: 17.

[0523] Embodiment 44 is the macrophage CAR of embodiment 9, wherein R16 comprises, consists essentially of, or consists of SEQ ID NO: 23.

[0524] Embodiment 45 is the macrophage CAR of embodiment 9, wherein R17 comprises, consists essentially of, or consists of SEQ ID NO: 25.

[0525] Embodiment 46 is the macrophage CAR of embodiment 10, wherein R18 comprises, consists essentially of, or consists of SEQ ID NO: 23.

[0526] Embodiment 47 is the macrophage CAR of embodiment 10, wherein R19 comprises, consists essentially of, or consists of SEQ ID NO: 25.

[0527] Embodiment 48 is the macrophage CAR of embodiment 11, wherein R20 comprises, consists essentially of, or consists of SEQ ID NO: 27.

[0528] Embodiment 49 is the macrophage CAR of embodiment 12, wherein R22 comprises, consists essentially of, or consists of SEQ ID NO: 33.

[0529] Embodiment 50 is the macrophage CAR of embodiment 13, wherein R23 comprises, consists essentially of, or consists of SEQ ID NO: 27.

[0530] Embodiment 51 is the macrophage CAR of embodiment 14, wherein R24 comprises, consists essentially of, or consists of SEQ ID NO: 33.

[0531] Embodiment 52 is the macrophage CAR of embodiment 19, wherein R27 comprises, consists essentially of, or consists of SEQ ID NO: 35.Embodiment 53 is the macrophage CAR of embodiment 20, wherein R28 comprises, consists essentially of, or consists of SEQ ID NO: 35.

[0532] Embodiment 54 is the macrophage CAR of embodiment 15 or embodiment 16, wherein R29 comprises, consists essentially of, or consists of SEQ ID NO: 37.

[0533] Embodiment 55 is the macrophage CAR of embodiment 15 or embodiment 16, wherein R30 comprises, consists essentially of, or consists of SEQ ID NO: 39.

[0534] Embodiment 56 is the macrophage CAR of embodiment 17 or embodiment 18, wherein R31 comprises, consists essentially of, or consists of SEQ ID NO: 37.

[0535] Embodiment 57 is the macrophage CAR of embodiment 17 or embodiment 18, wherein R32 comprises, consists essentially of, or consists of SEQ ID NO: 39.

[0536] Embodiment 58 is the macrophage CAR of embodiment 12, wherein R22 comprises, consists essentially of, or consists of SEQ ID NO: 33.

[0537] Embodiment 59 is the macrophage CAR of embodiment 12, wherein R22 comprises, consists essentially of, or consists of SEQ ID NO: 33.

[0538] Embodiment 60 is the macrophage CAR of embodiment 12, wherein R22 comprises, consists essentially of, or consists of SEQ ID NO: 33.

[0539] Embodiment 61 is a macrophage CAR comprising SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, or SEQ ID NO: 63.

[0540] Embodiment 62 is a vector for engineering a macrophage, wherein the vector is a retroviral vector, comprising a nucleic acid encoding the macrophage CAR of any one of embodiments 1-61.

[0541] Embodiment 63 is the vector of embodiment 62, wherein the retroviral vector is a lentiviral vector.

[0542] Embodiment 64 is an engineered macrophage, comprising the CAR of any one of embodiments 1-61 or the vector of embodiment 62 or 63.

[0543] Embodiment 65 is a pharmaceutical composition, comprising: the engineered macrophage of embodiment 64; and a pharmaceutically acceptable carrier.

[0544] Embodiment 66 is a method of treating cancer, comprising administering to a subject with cancer: the engineered macrophage of embodiment 64; or the pharmaceutical composition of embodiment 65.

[0545] Embodiment 67 is the method of embodiment 66, wherein the cancer is ALK-positive.

[0546] Embodiment 68 is the method of embodiment 66 or embodiment 67, wherein the cancer is selected from the list consisting of: neuroblastoma, non-small cell lung cancer, anaplastic large celllymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma.

[0547] Embodiment 69 is the method of embodiment 66, wherein the cancer is HER2 -positive. Embodiment 70 is the method of embodiment 66 or embodiment 69, wherein the cancer is selected from the list consisting of: breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer.

[0548] Embodiment 71 is an engineered macrophage of embodiment 64, for use in therapy, in particular for treating cancer.

[0549] Embodiment 72 is an engineered macrophage of embodiment 64, for use in treating cancer. Embodiment 73 is the engineered macrophage of embodiment 71 or embodiment 72, wherein the cancer is ALK-positive cancer.

[0550] Embodiment 74 is the engineered macrophage of any one of embodiments 71-73, wherein the cancer is selected from the list consisting of: neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma.

[0551] Embodiment 75 is the engineered macrophage of embodiment 71 or embodiment 72, wherein the cancer is HER2-positive cancer.

[0552] Embodiment 76 is the engineered macrophage of any one of embodiments 71-72 or embodiment 75, wherein the cancer is selected from the list consisting of: breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer.

[0553] Embodiment 77 is a use of the engineered macrophage of embodiment 64 in the manufacture of a medicament for treating cancer.

[0554] Embodiment 78 is the use of embodiment 77, wherein the cancer is ALK-positive cancer. Embodiment 79 is the use of embodiment 77 or embodiment 78, wherein the cancer is selected from the list consisting of: neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma.Embodiment 80 is the use of embodiment 77, wherein the cancer is HER2 -positive cancer. Embodiment 81 is the use of embodiment 77 or embodiment 80, wherein the cancer is selected from the list consisting of breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer.

[0555] Embodiment 82 is a pharmaceutical composition for the treatment of cancer, comprising: the engineered macrophage of embodiment 64; or the pharmaceutical composition of embodiment 65.

[0556] Embodiment 83 is the pharmaceutical composition of embodiment 82, wherein the cancer is ALK-positive.

[0557] Embodiment 84 is the pharmaceutical composition of embodiment 82 or embodiment 83, wherein the cancer is selected from the list consisting of: neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma.

[0558] Embodiment 85 is the pharmaceutical composition of embodiment 82, wherein the cancer is HER2-positive.

[0559] Embodiment 86 is the pharmaceutical composition of embodiment 82 or embodiment 85, wherein the HER2-positive cancer is selected from the list consisting of: breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer.

[0560] SEQUENCE LISTING FREE TEXT SEQ ID NO Sequence

[0561] 1 MDFQVQIFSFLLISASVIMSRGDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQ KPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQ GTKVEIKRTGSTSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHW VRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVY YCSRWGGDGFYAMDVWGQGTLVTVSS

[0562] 2 ATGGACTTCCAGGTGCAGATTTTCAGCTTCCTGCTGATCAGCGCCTCTGTGATCATGA GCCGGGGCGATATCCAGATGACACAGAGCCCATCTTCTCTGAGCGCCAGCGTGGGCG ATAGAGTGACAATCACATGTAGAGCCTCCCAGGATGTGAACACCGCTGTGGCCTGGT ATCAGCAGAAGCCTGGAAAAGCCCCTAAGCTGCTGATCTACAGCGCCTCTTTTCTCTA CAGCGGTGTTCCTAGCAGATTCTCCGGAAGCAGAAGCGGCACCGATTTTACCCTGAC CATTTCTAGCCTCCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGCACTACACA ACCCCTCCTACATTCGGCCAAGGCACCAAGGTGGAAATCAAGAGAACCGGCTCCACC AGCGGCTCCGGGAAGCCCGGCTCCGGAGAGGGCTCTGAAGTGCAACTGGTGGAAAG CGGCGGCGGCCTGGTGCAGCCTGGCGGCTCCCTGCGGCTGTCCTGCGCCGCTTCTGGT TTTAACATCAAGGACACATACATCCATTGGGTGCGGCAGGCCCCTGGCAAAGGCCTG GAATGGGTGGCCAGAATCTACCCCACAAACGGCTATACCAGATACGCCGATAGCGTG

[0563]

[0564] AAGGGCCGGTTCACCATCAGCGCCGACACCAGCAAGAACACAGCTTATCTGCAGATGAACAGCCTGAGAGCTGAGGATACCGCCGTGTACTACTGCAGCAGATGGGGCGGCGA CGGCTTCTACGCtATGGACGTGTGGGGACAGGGCACCCTGGTGACCGTGTCCAGC MLLLVTSLLLCELPHPAFLLIPQVQLQQSGAELAKPGASVKLSCKASGYTFTSYWMHWV KQRPGQGLEWIGYIKPSSGYTKYNQKFKDKATLTADKSSSTAYMQLSSLTYEDSAVYYC ARDYYGSSSWFAYWGQGTLVTVSAGGGGSGGGGSDIVMTQSQRFMSTSVGDRVSVTCK ASQNVGTNVAWYQQKPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLA EYFCQQYNSYPYMYTFGGGTKLE ATGCTCTTGTTGGTGACGAGTCTCCTGCTGTGTGAACTGCCGCACCCAGCATTTCTTTT GATTCCGCAGGTTCAGCTGCAGCAGTCTGGTGCCGAACTGGCTAAACCTGGCGCCTC TGTGAAGCTGAGCTGTAAAGCCAGCGGCTACACCTTCACCAGCTACTGGATGCACTG GGTCAAGCAGAGGCCTGGACAGGGACTCGAGTGGATCGGCTACATCAAGCCTAGCA GCGGCTATACCAAGTACAACCAGAAGTTCAAGGACAAGGCCACACTGACCGCCGAC AAGAGCAGCAGCACAGCCTACATGCAGCTGAGCAGCCTGACCTACGAGGACAGCGC CGTGTACTACTGCGCCAGAGATTACTACGGCAGCAGCAGTTGGTTCGCCTATTGGGG CCAGGGCACACTGGTTACAGTTTCTGCTGGTGGCGGAGGATCTGGCGGAGGCGGATC TGATATCGTGATGACCCAGAGCCAGCGGTTCATGAGCACAAGCGTGGGCGATAGAGT GTCCGTGACATGCAAGGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCA GAAGCCTGGGCAGTCTCCCAAGGCTCTGATCTACAGCGCCAGCTACAGATACAGCGG CGTGCCCGATAGATTCACAGGCAGCGGCTCTGGCACCGACTTCACCCTGACAATCAG CAACGTGCAGAGCGAGGACCTGGCCGAGTACTTCTGCCAGCAGTACAACAGCTACCC CTACATGTACACCTTCGGCGGAGGCACCAAGCTGGAA MALPVTALLLPLALLLHAARPQVQLQQSGAELAKPGASVKLSCKASGYTFTSYWMHWV KQRPGQGLEWIGYIKPSSGYTKYNQKFKDKATLTADKSSSTAYMQLSSLTYEDSAVYYC ARDYYGSSSWFAYWGQGTLVTVSAGGGGSGGGGSDIVMTQSQRFMSTSVGDRVSVTCK ASQNVGTNVAWYQQKPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLA EYFCQQYNSYPYMYTFGGGTKLE ATGGCTCTGCCTGTTACAGCTCTGCTGCTGCCTCTGGCTCTGCTTCTGCATGCTGCTAG ACCTCAGGTTCAGCTGCAGCAGTCTGGTGCCGAACTGGCTAAACCTGGCGCCTCTGT GAAGCTGAGCTGTAAAGCCAGCGGCTACACCTTCACCAGCTACTGGATGCACTGGGT CAAGCAGAGGCCTGGACAGGGACTCGAGTGGATCGGCTACATCAAGCCTAGCAGCG GCTATACCAAGTACAACCAGAAGTTCAAGGACAAGGCCACACTGACCGCCGACAAG AGCAGCAGCACAGCCTACATGCAGCTGAGCAGCCTGACCTACGAGGACAGCGCCGT GTACTACTGCGCCAGAGATTACTACGGCAGCAGCAGTTGGTTCGCCTATTGGGGCCA GGGCACACTGGTTACAGTTTCTGCTGGTGGCGGAGGATCTGGCGGAGGCGGATCTGA TATCGTGATGACCCAGAGCCAGCGGTTCATGAGCACAAGCGTGGGCGATAGAGTGTC CGTGACATGCAAGGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGA AGCCTGGGCAGTCTCCCAAGGCTCTGATCTACAGCGCCAGCTACAGATACAGCGGCG TGCCCGATAGATTCACAGGCAGCGGCTCTGGCACCGACTTCACCCTGACAATCAGCA ACGTGCAGAGCGAGGACCTGGCCGAGTACTTCTGCCAGCAGTACAACAGCTACCCCT ACATGTACACCTTCGGCGGAGGCACCAAGCTGGAA LDTFPYKCLNSLQVLDYSLNHIMTSKKQELQHFPSSLAFLNLTQNDFACTCEHQSFLQWI KDQRQLLVEVERMECATPSDKQGMPVLSLN TTGGATACGTTTCCTTATAAGTGTCTGAACTCCCTCCAGGTTCTTGATTACAGTCTCAA TCACATAATGACTTCCAAAAAACAGGAACTACAGCATTTTCCAAGTAGTCTAGCTTTC TTAAATCTTACTCAGAATGACTTTGCTTGTACTTGTGAACACCAGAGTTTCCTGCAAT GGATCAAGGACCAGAGGCAGCTCTTGGTGGAAGTTGAACGAATGGAATGTGCAACA CCTTCAGATAAGCAGGGCATGCCTGTGCTGAGTTTGAAT ITCQMNKTIIGVSVLSVLWSWAVLVYKFYFHL ATCACCTGTCAGATGAATAAGACCATCATTggtgtgtcggtcctcagtgtgcttgtagtatctgttgtagcagttctg gtctataagttctattttcacctg MLLAGCIKYGRGENIYDAFVIYSSQDEDWVRNELVKNLEEGVPPFQLCLHYRDFIPGVAI AANIIHEGFHKSRKVIVWSQHFIQSRWCIFEYEIAQTWQFLSSRAGIIFIVLQKVEKTLLRQ QVELYRLLSRNTYLEWEDSVLGRHIFWRRLRKALLDG

[0565] atgctcttgctggctgcataaagtatggtagaggtgaaaacatctatgatgcctttgttatctactcaagccaggatgaggactgggtaaggaat gagctagtaaagaatttagaagaaggggtgcctccatttcagctctgccttcactacagagactttattcccggtgtggccattgctgccaacat catccatgaaggtttccataaaagccgaaaggtgattgtgtggtgtcccagcactcatccagagccgctggtgtatctttgaatatgagattgc

[0566]

[0567] tcagacctggcagtttctgagcagtcgtgctggtatcatcttcattgtcctgcagaaggtggagaagaccctgctcaggcagcaggtggagctgtaccgccttctcagcaggaacacttacctggagtgggaggacagtgtcctggggcggcacatcttctggagacgactcagaaaagccctg ctggatggt LNSLQVLDYSLNHIMTSKKQELQHFPSSLAFLNLTQNDFACTCEHQSFLQWIKDQRQLLV EVERMECATPSDKQGMPVLSLN CTGAACTCCCTCCAGGTTCTTGATTACAGTCTCAATCACATAATGACTTCCAAAAAAC AGGAACTACAGCATTTTCCAAGTAGTCTAGCTTTCTTAAATCTTACTCAGAATGACTT TGCTTGTACTTGTGAACACCAGAGTTTCCTGCAATGGATCAAGGACCAGAGGCAGCT CTTGGTGGAAGTTGAACGAATGGAATGTGCAACACCTTCAGATAAGCAGGGCATGCC TGTGCTGAGTTTGAAT IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSK ATCGAGGTGATGTACCCTCCACCATACCTGGACAATGAGAAGTCCAATGGTACCATC ATCCATGTGAAGGGCAAGCACCTGTGTCCTAGCCCCCTGTTCCCCGGCCCCTCTAAA PFWVLVWGGVLACYSLLVTVAFIIFWV CCTTTCTGGGTCCTGGTGGTGGTCGGAGGCGTGCTGGCCTGCTACAGCCTGCTGGTCA CCGTTGCCTTCATCATCTTTTGGGTG LVLKYRGLIKYWFHTPPSIPLQIEEYLKDPTQPILEALDKDSSPKDDVWDSVSIISFPEKEQE DVLQTLGGGGSGGGGSGGGGSCFYIKKINPLKEKSIILPKSLISVVRSATLETKPESKYVSL ITSYQPFSLEKEWCEEPLSPATVPGMHTEDNPGKVEHTEELSSITEWTTEENIPDVVPGS HLTPIERESSSPLSSNQSEPGSIALNSYHSRNCSESDHSRNGFDTDSSCLESHSSLSDSEFPPN NKGEIKTEGQELITVIKAPTSFGYDKPHVLVDLLVDDSGKESLIGYRPTEDSKEFS

[0568] ctggtcctgaaatatagaggcctgattaaatactggttcacactccaccaagcatcccattacagatagaagagtatttaaaagacccaactca gcccatcttagaggccttggacaaggacagctcaccaaaggatgacgtctgggactctgtgtccattatctcgtttccggaaaaggagcaaga agatgttctccaaacgcttGGTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGATCTt gtttttatattaagaaaattaatccattgaaggaaaaaagcataatattacccaagtccttgatctctgtggtaagaagtgctacttagagacaaaa cctgaatcaaaatatgtatcactcatcacgtcataccagccattttccttagaaaaggaggtggtctgtgaagagccgtlgtctccagcaacagtt ccaggcatgcataccgaagacaatccaggaaaagtggaacatacagaagaactttctagtataacagaagtggtgactactgaagaaaatatt cctgacgtggtcccgggcagccatctgactccaatagagagagagagttcttcacctttaagtagtaaccagtctgaacctggcagcatcgctt taaactcgtatcactccagaaattgttctgagagtgatcactccagaaatggtttgatactgattccagctgtctggaatcacatagctccttatct gactcagaatttcccccaaataataaaggtgaaataaaaacagaaggacaagagctcataaccgtaataaaagcccccacctcctttggttatg ataaaccacatgtgctagtggatctactgtggatgatagcggtaaagagtccttgattggttatagaccaacagaagattccaaagaatttca CFYIKKINPLKEKSIILPKSLISWRSATLETKPESKYVSLITSYQPFSLEKEWCEEPLSPAT VPGMHTEDNPGKVEHTEELSS1TEVVTTEEN1PDVVPGSHLTPIERESSSPLSSNQSEPGSIA LNSYHSRNCSESDHSRNGFDTDSSCLESHSSLSDSEFPPNNKGEIKTEGQELITVIKAPTSFG YDKPHVLVDLLVDDSGKESLIGYRPTEDSKEFSGGGGSGGGGSGGGGSGGGGSLVLKYR GLIKYWFHTPPSIPLQIEEYLKDPTQPILEALDKDSSPKDDVWDSVSIISFPEKEQEDVLQTL

[0569] tgtttttatataagaaaattaatccatgaaggaaaaaagcataatatacccaagtccttgatctctgtggtaagaagtgctactttagagacaaa acctgaatcaaaatatgtatcactcatcacgtcataccagccatttccttagaaaaggaggtggtctgtgaagagccgttgtctccagcaacag ttccaggcatgcataccgaagacaatccaggaaaagtggaacatacagaagaactttctagtataacagaagtggtgactactgaagaaaata ttcctgacgtggtcccgggcagccatctgactccaatagagagagagagtctcacctttaagtagtaaccagtctgaacctggcagcatcgc tttaaactcgtatcactccagaaattgttctgagagtgatcactccagaaatggttttgatactgattccagctgtctggaatcacatagctccttatc tgactcagaatttcccccaaataataaaggtgaaataaaaacagaaggacaagagctcataaccgtaataaaagcccccacctcctttggttat gataaaccacatgtgctagtggatctacttgtggatgatagcggtaaagagtccttgattggttatagaccaacagaagattccaaagaatttca GGTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGG ATCTctggtcctgaaatatagaggcctgattaaatactggtttcacactccaccaagcatcccattacagatagaagagtatttaaaagaccc aactcagcccatcttagaggccttggacaaggacagctcaccaaaggatgacgtctgggactctgtgtccattatctcgtttccggaaaagga gcaagaagatgttctccaaacgctt ALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEASRPAAGGAVHTRGLD VDA GCCCTGAGCAACAGCATCATGTACTTCAGCCACTTCGTGCCCGTGTTTCTGCCCGCCA AGCCTACAACAACCCCTGCTCCTAGACCACCTACACCAGCTCCTACAATCGCCAGCC AGCCTCTGTCTCTGAGGCCTGAAGCTTCTAGACCAGCTGCTGGCGGAGCCGTGCATA CCAGAGGACTTGATGTCGACGCT SSLWIPWAALLLFLVLSLVFI AGCtctctttggattccagttgttgctgctttactactctttctagtgcttagcctggtattcatc

[0570] MDFQVQIFSFLL1SASVIMSRG

[0571]

[0572] ATGGACTTCCAGGTGCAGATTTTCAGCTTCCTGCTGATCAGCGCCTCTGTGATCATGA GCCGGGGC DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTGSTSGSGKPGSGE GSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGY TRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDVWGQGT LVTVSS GATATCCAGATGACACAGAGCCCATCTTCTCTGAGCGCCAGCGTGGGCGATAGAGTG ACAATCACATGTAGAGCCTCCCAGGATGTGAACACCGCTGTGGCCTGGTATCAGCAG AAGCCTGGAAAAGCCCCTAAGCTGCTGATCTACAGCGCCTCTTTTCTCTACAGCGGTG TTCCTAGCAGATTCTCCGGAAGCAGAAGCGGCACCGATTTTACCCTGACCATTTCTAG CCTCCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGCACTACACAACCCCTCCT ACATTCGGCCAAGGCACCAAGGTGGAAATCAAGAGAACCGGCTCCACCAGCGGCTC CGGGAAGCCCGGCTCCGGAGAGGGCTCTGAAGTGCAACTGGTGGAAAGCGGCGGCG GCCTGGTGCAGCCTGGCGGCTCCCTGCGGCTGTCCTGCGCCGCTTCTGGTTTTAACAT CAAGGACACATACATCCATTGGGTGCGGCAGGCCCCTGGCAAAGGCCTGGAATGGGT GGCCAGAATCTACCCCACAAACGGCTATACCAGATACGCCGATAGCGTGAAGGGCCG GTTCACCATCAGCGCCGACACCAGCAAGAACACAGCTTATCTGCAGATGAACAGCCT GAGAGCTGAGGATACCGCCGTGTACTACTGCAGCAGATGGGGCGGCGACGGCTTCTA CGCtATGGACGTGTGGGGACAGGGCACCCTGGTGACCGTGTCCAGC QVQLQQSGAELAKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGYIKPSSGYT KYNQKFKDKATLTADKSSSTAYMQLSSLTYEDSAVYYCARDYYGSSSWFAYWGQGTL VTVSAGGGGSGGGGSDIVMTQSQRFMSTSVGDRVSVTCKASQNVGTNVAWYQQKPGQ SPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLAEYFCQQYNSYPYMYTFGG GTKLE CAGGTTCAGCTGCAGCAGTCTGGTGCCGAACTGGCTAAACCTGGCGCCTCTGTGAAG CTGAGCTGTAAAGCCAGCGGCTACACCTTCACCAGCTACTGGATGCACTGGGTCAAG CAGAGGCCTGGACAGGGACTCGAGTGGATCGGCTACATCAAGCCTAGCAGCGGCTAT ACCAAGTACAACCAGAAGTTCAAGGACAAGGCCACACTGACCGCCGACAAGAGCAG CAGCACAGCCTACATGCAGCTGAGCAGCCTGACCTACGAGGACAGCGCCGTGTACTA CTGCGCCAGAGATTACTACGGCAGCAGCAGTTGGTTCGCCTATTGGGGCCAGGGCAC ACTGGTTACAGTTTCTGCTGGTGGCGGAGGATCTGGCGGAGGCGGATCTGATATCGT GATGACCCAGAGCCAGCGGTTCATGAGCACAAGCGTGGGCGATAGAGTGTCCGTGAC ATGCAAGGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGAAGCCTG GGCAGTCTCCCAAGGCTCTGATCTACAGCGCCAGCTACAGATACAGCGGCGTGCCCG ATAGATTCACAGGCAGCGGCTCTGGCACCGACTTCACCCTGACAATCAGCAACGTGC AGAGCGAGGACCTGGCCGAGTACTTCTGCCAGCAGTACAACAGCTACCCCTACATGT ACACCTTCGGCGGAGGCACCAAGCTGGAA MLLLVTSLLLCELPHPAFLLIP ATGCTCTTGTTGGTGACGAGTCTCCTGCTGTGTGAACTGCCGCACCCAGCATTTCTTTT GATTCCG MALPVTALLLPLALLLHAARP ATGGCTCTGCCTGTTACAGCTCTGCTGCTGCCTCTGGCTCTGCTTCTGCATGCTGCTAG ACCT CFYIKKINPLKEKSIILPKSLISWRSATLETKPESKYVSLITSYQPFSLEKEWCEEPLSPAT VPGMHTEDNPGKVEHTEELSSITEWTTEENIPDWPGSHLTPIERESSSPLSSNQSEPGSIA LNSYHSRNCSESDHSRNGFDTDSSCLESHSSLSDSEFPPNNKGEIKTEGQELITVIKAPTSFG YDKPHVLVDLLVDDSGKESLIGYRPTEDSKEFS

[0573] tgttttatattaagaaaattaatccattgaaggaaaaaagcataatattacccaagtccttgatctctgtggtaagaagtgctactttagagacaaa acctgaatcaaaatatgtatcactcatcacgtcataccagccattttccttagaaaaggaggtggtctgtgaagagccgttgtctccagcaacag ttccaggcatgcataccgaagacaatccaggaaaagtggaacatacagaagaactttctagtataacagaagtggtgactactgaagaaaata ttcctgacgtggtcccgggcagccatctgactccaatagagagagagagttcttcacctttaagtagtaaccagtctgaacctggcagcatcgc ttaaactcgtatcactccagaaattgttctgagagtgatcactccagaaatggttttgatactgatccagctgtctggaatcacatagctcctatc tgactcagaatttcccccaaataataaaggtgaaataaaaacagaaggacaagagctcataaccgtaataaaagcccccacctcctttggttat gataaaccacatgtgctagtggatctacttgtggatgatagcggtaaagagtccttgattggttatagaccaacagaagattccaaagaatttca LVLKYRGLIKYWFHTPPSIPLQIEEYLKDPTQPILEALDKDSSPKDDVWDSVSIISFPEKEQE

[0574]

[0575] DVLQTLctggtcctgaaatatagaggcctgattaaatactggtttcacactccaccaagcatcccattacagatagaagagtatttaaaagacccaactca gcccatcttagaggcctggacaaggacagctcaccaaaggatgacgtctgggactctgtgtccattatctcgtttccggaaaaggagcaaga agatgttctccaaacgct

[0576] GGGGS GGGS GGGGSGGGGSGGGGS GGTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGATCT GGGGSGGGGSGGGGSGGGGS GGTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGG ATCT DYKDDDDKAAA GATTACAAGGACGACGATGACAAGGCCGCCGCC MDFQVQIFSFLLISASVIMSRGDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQ KPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQ GTKVEIKRTGSTSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHW VRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVY YCSRWGGDGFYAMDVWGQGTLVTVSSDYKDDDDKAAALDTFPYKCLNSLQVLDYSLN

[0577] H1MTSKKQELQHFPSSLAFLNLTQNDFACTCEHQSFLQWIKDQRQLLVEVERMECATPSD KQGMPVLSLNITCQMNKTIIGVSVLSVLWSWAVLVYKFYFHLMLLAGCIKYGRGENIY DAFVIYSSQDEDWVRNELVKNLEEGVPPFQLCLHYRDFIPGVAIAANIIHEGFHKSRKVIV WSQHFIQSRWCIFEYEIAQTWQFLSSRAGIIFIVLQKVEKTLLRQQVELYRLLSRNTYLE WEDSVLGRHIFWRRLRKALLDG ATGGACTTCCAGGTGCAGATTTTCAGCTTCCTGCTGATCAGCGCCTCTGTGATCATGA GCCGGGGCGATATCCAGATGACACAGAGCCCATCTTCTCTGAGCGCCAGCGTGGGCG ATAGAGTGACAATCACATGTAGAGCCTCCCAGGATGTGAACACCGCTGTGGCCTGGT ATCAGCAGAAGCCTGGAAAAGCCCCTAAGCTGCTGATCTACAGCGCCTCTTTTCTCTA CAGCGGTGTTCCTAGCAGATTCTCCGGAAGCAGAAGCGGCACCGATTTTACCCTGAC CATTTCTAGCCTCCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGCACTACACA ACCCCTCCTACATTCGGCCAAGGCACCAAGGTGGAAATCAAGAGAACCGGCTCCACC AGCGGCTCCGGGAAGCCCGGCTCCGGAGAGGGCTCTGAAGTGCAACTGGTGGAAAG CGGCGGCGGCCTGGTGCAGCCTGGCGGCTCCCTGCGGCTGTCCTGCGCCGCTTCTGGT TTTAACATCAAGGACACATACATCCATTGGGTGCGGCAGGCCCCTGGCAAAGGCCTG GAATGGGTGGCCAGAATCTACCCCACAAACGGCTATACCAGATACGCCGATAGCGTG AAGGGCCGGTTCACCATCAGCGCCGACACCAGCAAGAACACAGCTTATCTGCAGATG AACAGCCTGAGAGCTGAGGATACCGCCGTGTACTACTGCAGCAGATGGGGCGGCGA CGGCTTCTACGCtATGGACGTGTGGGGACAGGGCACCCTGGTGACCGTGTCCAGCGAT TACAAGGACGACGATGACAAGGCCGCCGCCTTGGATACGTTTCCTTATAAGTGTCTG AACTCCCTCCAGGTTCTTGATTACAGTCTCAATCACATAATGACTTCCAAAAAACAGG AACTACAGCATTTTCCAAGTAGTCTAGCTTTCTTAAATCTTACTCAGAATGACTTTGCT TGTACTTGTGAACACCAGAGTTTCCTGCAATGGATCAAGGACCAGAGGCAGCTCTTG GTGGAAGTTGAACGAATGGAATGTGCAACACCTTCAGATAAGCAGGGCATGCCTGTG CTGAGTTTGAATATCACCTGTCAGATGAATAAGACCATCATTggtgtgtcggtcctcagtgtgcttgta gtatctgttgtagcagttctggtctataagttctattttcacctgatgcttcttgctggctgcataaagtatggtagaggtgaaaacatctatgatgcc ttgttatctactcaagccaggatgaggactgggtaaggaatgagctagtaaagaatttagaagaaggggtgcctccatttcagctctgccttca ctacagagacttattcccggtgtggccattgctgccaacatcatccatgaaggtttccataaaagccgaaaggtgattgttgtggtgtcccagc acttcatccagagccgctggtgtatctttgaatatgagattgctcagacctggcagtttctgagcagtcgtgctggtatcatcttcattgtcctgca gaaggtggagaagaccctgctcaggcagcaggtggagctgtaccgccttctcagcaggaacacttacctggagtgggaggacagtgtcct ggggcggcacatcttctggagacgactcagaaaagccctgctggatggttag MDFQVQIFSFLLISASVIMSRGDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQ KPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQ GTKVEIKRTGSTSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHW VRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVY YCSRWGGDGFYAMDVWGQGTLVTVSSDYKDDDDKAAALNSLQVLDYSLNHIMTSKKQ ELQHFPSSLAFLNLTQNDFACTCEHQSFLQWIKDQRQLLVEVERMECATPSDKQGMPVL SLNITCQMNKTIIGVSVLSVLWSWAVLVYKFYFHLMLLAGCIKYGRGENIYDAFVIYSS

[0578]

[0579] QDEDWVRNELVKNLEEGVPPFQLCLHYRDFIPGVAIAANIIHEGFHKSRKVIVWSQHFIQSRWCIFEYEIAQTWQFLSSRAGIIFIVLQKVEKTLLRQQVELYRLLSRNTYLEWEDSVLGR HIFWRRLRKALLDG ATGGACTTCCAGGTGCAGATTTTCAGCTTCCTGCTGATCAGCGCCTCTGTGATCATGA GCCGGGGCGATATCCAGATGACACAGAGCCCATCTTCTCTGAGCGCCAGCGTGGGCG ATAGAGTGACAATCACATGTAGAGCCTCCCAGGATGTGAACACCGCTGTGGCCTGGT ATCAGCAGAAGCCTGGAAAAGCCCCTAAGCTGCTGATCTACAGCGCCTCTTTTCTCTA CAGCGGTGTTCCTAGCAGATTCTCCGGAAGCAGAAGCGGCACCGATTTTACCCTGAC CATTTCTAGCCTCCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGCACTACACA ACCCCTCCTACATTCGGCCAAGGCACCAAGGTGGAAATCAAGAGAACCGGCTCCACC AGCGGCTCCGGGAAGCCCGGCTCCGGAGAGGGCTCTGAAGTGCAACTGGTGGAAAG CGGCGGCGGCCTGGTGCAGCCTGGCGGCTCCCTGCGGCTGTCCTGCGCCGCTTCTGGT TTTAACATCAAGGACACATACATCCATTGGGTGCGGCAGGCCCCTGGCAAAGGCCTG GAATGGGTGGCCAGAATCTACCCCACAAACGGCTATACCAGATACGCCGATAGCGTG AAGGGCCGGTTCACCATCAGCGCCGACACCAGCAAGAACACAGCTTATCTGCAGATG AACAGCCTGAGAGCTGAGGATACCGCCGTGTACTACTGCAGCAGATGGGGCGGCGA CGGCTTCTACGCtATGGACGTGTGGGGACAGGGCACCCTGGTGACCGTGTCCAGCGAT TACAAGGACGACGATGACAAGGCCGCCGCCCTGAACTCCCTCCAGGTTCTTGATTAC AGTCTCAATCACATAATGACTTCCAAAAAACAGGAACTACAGCATTTTCCAAGTAGT CTAGCTTTCTTAAATCTTACTCAGAATGACTTTGCTTGTACTTGTGAACACCAGAGTTT CCTGCAATGGATCAAGGACCAGAGGCAGCTCTTGGTGGAAGTTGAACGAATGGAATG TGCAACACCTTCAGATAAGCAGGGCATGCCTGTGCTGAGTTTGAATATCACCTGTCAG ATGAATAAGACCATCATTggtgtgtcggtcctcagtgtgctgtagtatctgtgtagcagttctggtctataagttctatttcac ctgatgcttcttgctggctgcataaagtatggtagaggtgaaaacatctatgatgcctttgttatctactcaagccaggatgaggactgggtaag gaatgagctagtaaagaatttagaagaaggggtgcctccatttcagctctgccttcactacagagactttattcccggtgtggccattgctgcca acatcatccatgaaggtttccataaaagccgaaaggtgatgttgtggtgtcccagcacttcatccagagccgctggtgtatctttgaatatgaga ttgctcagacctggcagtttctgagcagtcgtgctggtatcatcttcattgtcctgcagaaggtggagaagaccctgctcaggcagcaggtgga gctgtaccgccttctcagcaggaacacttacctggagtgggaggacagtgtcctggggcggcacatctctggagacgactcagaaaagcc ctgctggatggttag MDFQVQ1FSFLL1SASV1MSRGDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQ KPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQ GTKVEIKRTGSTSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHW VRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVY YCSRWGGDGFYAMDVWGQGTLVTVSSDYKDDDDKAAAIEVMYPPPYLDNEKSNGTIIH VKGKHLCPSPLFPGPSKPFWVLWVGGVLACYSLLVTVAFIIFWVLVLKYRGLIKYWFHT PPSIPLQIEEYLKDPTQPILEALDKDSSPKDDVWDSVSIISFPEKEQEDVLQTLGGGGSGGG GSGGGGSCFYIKKINPLKEKSIILPKSLISWRSATLETKPESKYVSLITSYQPFSLEKEWCE EPLSPATVPGMHTEDNPGKVEHTEELSSITEVVTTEENIPDWPGSHLTPIERESSSPLSSNQ SEPGSIALNSYHSRNCSESDHSRNGFDTDSSCLESHSSLSDSEFPPNNKGEIKTEGQELITVI KAPTSFGYDKPHVLVDLLVDDSGKESLIGYRPTEDSKEFS ATGGACTTCCAGGTGCAGATTTTCAGCTTCCTGCTGATCAGCGCCTCTGTGATCATGA GCCGGGGCGATATCCAGATGACACAGAGCCCATCTTCTCTGAGCGCCAGCGTGGGCG ATAGAGTGACAATCACATGTAGAGCCTCCCAGGATGTGAACACCGCTGTGGCCTGGT ATCAGCAGAAGCCTGGAAAAGCCCCTAAGCTGCTGATCTACAGCGCCTCTTTTCTCTA CAGCGGTGTTCCTAGCAGATTCTCCGGAAGCAGAAGCGGCACCGATTTTACCCTGAC CATTTCTAGCCTCCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGCACTACACA ACCCCTCCTACATTCGGCCAAGGCACCAAGGTGGAAATCAAGAGAACCGGCTCCACC AGCGGCTCCGGGAAGCCCGGCTCCGGAGAGGGCTCTGAAGTGCAACTGGTGGAAAG CGGCGGCGGCCTGGTGCAGCCTGGCGGCTCCCTGCGGCTGTCCTGCGCCGCTTCTGGT TTTAACATCAAGGACACATACATCCATTGGGTGCGGCAGGCCCCTGGCAAAGGCCTG GAATGGGTGGCCAGAATCTACCCCACAAACGGCTATACCAGATACGCCGATAGCGTG AAGGGCCGGTTCACCATCAGCGCCGACACCAGCAAGAACACAGCTTATCTGCAGATG AACAGCCTGAGAGCTGAGGATACCGCCGTGTACTACTGCAGCAGATGGGGCGGCGA CGGCTTCTACGCtATGGACGTGTGGGGACAGGGCACCCTGGTGACCGTGTCCAGCGAT TACAAGGACGACGATGACAAGGCCGCCGCCATCGAGGTGATGTACCCTCCACCATAC CTGGACAATGAGAAGTCCAATGGTACCATCATCCATGTGAAGGGCAAGCACCTGTGT CCTAGCCCCCTGTTCCCCGGCCCCTCTAAACCTTTCTGGGTCCTGGTGGTGGTCGGAG GCGTGCTGGCCTGCTACAGCCTGCTGGTCACCGTTGCCTTCATCATCTTTTGGGTGctgg

[0580]

[0581] tcctgaaatatagaggcctgattaaatactggtttcacactccaccaagcatcccattacagatagaagagtatttaaaagacccaactcagcccatcttagaggccttggacaaggacagctcaccaaaggatgacgtctgggactctgtgtccattatctcgtttccggaaaaggagcaagaagat gttctccaaacgcttGGTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGATCTtgtttt atattaagaaaattaatccattgaaggaaaaaagcataatattacccaagtccttgatctctgtggtaagaagtgctactttagagacaaaacctg aatcaaaatatgtatcactcatcacgtcataccagccattttccttagaaaaggaggtggtctgtgaagagccgttgtctccagcaacagttcca ggcatgcataccgaagacaatccaggaaaagtggaacatacagaagaacttctagtataacagaagtggtgactactgaagaaaatattcct gacgtggtcccgggcagccatctgactccaatagagagagagagttcttcaccttaagtagtaaccagtctgaacctggcagcatcgctttaa actcgtatcactccagaaatgttctgagagtgatcactccagaaatggtttgatactgattccagctgtctggaatcacatagctccttatctgac tcagaattcccccaaataataaaggtgaaataaaaacagaaggacaagagctcataaccgtaataaaagcccccacctccttggttatgata aaccacatgtgctagtggatctacttgtggatgatagcggtaaagagtccttgattggttatagaccaacagaagattccaaagaattttcatga MDFQVQIFSFLLISASVIMSRGDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQ KPGKAPKLLTYSASFLYSGVPSRFSGSRSGTDFTLTTSSLQPEDFATYYCQQHYTTPPTFGQ GTKVEIKRTGSTSGSGKPGSGEGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHW VRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVY YCSRWGGDGFYAMDVWGQGTLVTVSSDYKDDDDKAAAIEVMYPPPYLDNEKSNGTIIH VKGKHLCPSPLFPGPSKPFWVLVWGGVLACYSLLVTVAFIIFWVCFYIKKINPLKEKSIIL PKSLISWRSATLETKPESKYVSLITSYQPFSLEKEWCEEPLSPATVPGMHTEDNPGKVEH TEELSSITEWTTEENIPDWPGSHLTPIERESSSPLSSNQSEPGSIALNSYHSRNCSESDHSR NGFDTDSSCLESHSSLSDSEFPPNNKGEIKTEGQELITVIKAPTSFGYDKPHVLVDLLVDDS GKESLIGYRPTEDSKEFSGGGGSGGGGSGGGGSGGGGSLVLKYRGLIKYWFHTPPSIPLQI EEYLKDPTQPILEALDKDSSPKDDVWDSVSIISFPEKEQEDVLQTL ATGGACTTCCAGGTGCAGATTTTCAGCTTCCTGCTGATCAGCGCCTCTGTGATCATGA GCCGGGGCGATATCCAGATGACACAGAGCCCATCTTCTCTGAGCGCCAGCGTGGGCG ATAGAGTGACAATCACATGTAGAGCCTCCCAGGATGTGAACACCGCTGTGGCCTGGT ATCAGCAGAAGCCTGGAAAAGCCCCTAAGCTGCTGATCTACAGCGCCTCTTTTCTCTA CAGCGGTGTTCCTAGCAGATTCTCCGGAAGCAGAAGCGGCACCGATTTTACCCTGAC CATTTCTAGCCTCCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGCACTACACA ACCCCTCCTACATTCGGCCAAGGCACCAAGGTGGAAATCAAGAGAACCGGCTCCACC AGCGGCTCCGGGAAGCCCGGCTCCGGAGAGGGCTCTGAAGTGCAACTGGTGGAAAG CGGCGGCGGCCTGGTGCAGCCTGGCGGCTCCCTGCGGCTGTCCTGCGCCGCTTCTGGT TTTAACATCAAGGACACATACATCCATTGGGTGCGGCAGGCCCCTGGCAAAGGCCTG GAATGGGTGGCCAGAATCTACCCCACAAACGGCTATACCAGATACGCCGATAGCGTG AAGGGCCGGTTCACCATCAGCGCCGACACCAGCAAGAACACAGCTTATCTGCAGATG AACAGCCTGAGAGCTGAGGATACCGCCGTGTACTACTGCAGCAGATGGGGCGGCGA CGGCTTCTACGCtATGGACGTGTGGGGACAGGGCACCCTGGTGACCGTGTCCAGCGAT TACAAGGACGACGATGACAAGGCCGCCGCCATCGAGGTGATGTACCCTCCACCATAC CTGGACAATGAGAAGTCCAATGGTACCATCATCCATGTGAAGGGCAAGCACCTGTGT CCTAGCCCCCTGTTCCCCGGCCCCTCTAAACCTTTCTGGGTCCTGGTGGTGGTCGGAG GCGTGCTGGCCTGCTACAGCCTGCTGGTCACCGTTGCCTTCATCATCTTTTGGGTGtgtttt tatattaagaaaattaatccattgaaggaaaaaagcataatatacccaagtccttgatctctgtggtaagaagtgctactttagagacaaaacct gaatcaaaatatgtatcactcatcacgtcataccagccatttccttagaaaaggaggtggtctgtgaagagccgttgtctccagcaacagttcc aggcatgcataccgaagacaatccaggaaaagtggaacatacagaagaactttctagtataacagaagtggtgactactgaagaaaatattcc tgacgtggtcccgggcagccatctgactccaatagagagagagagttcttcacctttaagtagtaaccagtctgaacctggcagcatcgctta aactcgtatcactccagaaattgttctgagagtgatcactccagaaatggttttgatactgattccagctgtctggaatcacatagctccttatctga ctcagaattcccccaaataataaaggtgaaataaaaacagaaggacaagagctcataaccgtaataaaagcccccacctcctttggtatgat aaaccacatgtgctagtggatctacttgtggatgatagcggtaaagagtccttgattggttatagaccaacagaagattccaaagaattttcaG GTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGA TCTctggtcctgaaatatagaggcctgattaaatactggtttcacactccaccaagcatcccattacagatagaagagtattaaaagaccca actcagcccatcttagaggccttggacaaggacagctcaccaaaggatgacgtctgggactctgtgtccattatctcgtttccggaaaaggag caagaagatgttctccaaacgcttga MLLLVTSLLLCELPHPAFLLIPQVQLQQSGAELAKPGASVKLSCKASGYTFTSYWMHWV KQRPGQGLEWIGYIKPSSGYTKYNQKFKDKATLTADKSSSTAYMQLSSLTYEDSAVYYC ARDYYGSSSWFAYWGQGTLVTVSAGGGGSGGGGSDIVMTQSQRFMSTSVGDRVSVTCK ASQNVGTNVAWYQQKPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLA EYFCQQYNSYPYMYTFGGGTKLELDTFPYKCLNSLQVLDYSLNHIMTSKKQELQHFPSSL AFLNLTQNDFACTCEHQSFLQWIKDQRQLLVEVERMECATPSDKQGMPVLSLNITCQMN KTIIGVSVLSVLWSWAVLVYKFYFHLMLLAGCIKYGRGENIYDAFVIYSSQDEDWVRN

[0582]

[0583] ELVKNLEEGVPPFQLCLHYRDFIPGVAIAANIIHEGFHKSRKVIVWSQHFIQSRWCIFEYEIAQTWQFLSSRAGIIFIVLQKVEKTLLRQQVELYRLLSRNTYLEWEDSVLGRHIFWRRLRK ALLDG ATGCTCTTGTTGGTGACGAGTCTCCTGCTGTGTGAACTGCCGCACCCAGCATTTCTTTT GATTCCGCAGGTTCAGCTGCAGCAGTCTGGTGCCGAACTGGCTAAACCTGGCGCCTC TGTGAAGCTGAGCTGTAAAGCCAGCGGCTACACCTTCACCAGCTACTGGATGCACTG GGTCAAGCAGAGGCCTGGACAGGGACTCGAGTGGATCGGCTACATCAAGCCTAGCA GCGGCTATACCAAGTACAACCAGAAGTTCAAGGACAAGGCCACACTGACCGCCGAC AAGAGCAGCAGCACAGCCTACATGCAGCTGAGCAGCCTGACCTACGAGGACAGCGC CGTGTACTACTGCGCCAGAGATTACTACGGCAGCAGCAGTTGGTTCGCCTATTGGGG CCAGGGCACACTGGTTACAGTTTCTGCTGGTGGCGGAGGATCTGGCGGAGGCGGATC TGATATCGTGATGACCCAGAGCCAGCGGTTCATGAGCACAAGCGTGGGCGATAGAGT GTCCGTGACATGCAAGGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCA GAAGCCTGGGCAGTCTCCCAAGGCTCTGATCTACAGCGCCAGCTACAGATACAGCGG CGTGCCCGATAGATTCACAGGCAGCGGCTCTGGCACCGACTTCACCCTGACAATCAG CAACGTGCAGAGCGAGGACCTGGCCGAGTACTTCTGCCAGCAGTACAACAGCTACCC CTACATGTACACCTTCGGCGGAGGCACCAAGCTGGAATTGGATACGTTTCCTTATAAG TGTCTGAACTCCCTCCAGGTTCTTGATTACAGTCTCAATCACATAATGACTTCCAAAA AACAGGAACTACAGCATTTTCCAAGTAGTCTAGCTTTCTTAAATCTTACTCAGAATGA CTTTGCTTGTACTTGTGAACACCAGAGTTTCCTGCAATGGATCAAGGACCAGAGGCA GCTCTTGGTGGAAGTTGAACGAATGGAATGTGCAACACCTTCAGATAAGCAGGGCAT GCCTGTGCTGAGTTTGAATATCACCTGTCAGATGAATAAGACCATCATTggtgtgtcggtcctc agtgtgcttgtagtatctgttgtagcagttctggtctataagttctattttcacctgatgcttcttgctggctgcataaagtatggtagaggtgaaaac atctatgatgcctttgttatctactcaagccaggatgaggactgggtaaggaatgagctagtaaagaatttagaagaaggggtgcctccatttca gctctgccttcactacagagacttattcccggtgtggccattgctgccaacatcatccatgaaggttccataaaagccgaaaggtgattgttgt ggtgtcccagcacttcatccagagccgctggtgtatctttgaatatgagattgctcagacctggcagttctgagcagtcgtgctggtatcatctt cattgtcctgcagaaggtggagaagaccctgctcaggcagcaggtggagctgtaccgccttctcagcaggaacacttacctggagtgggag gacagtgtcctggggcggcacatcttctggagacgactcagaaaagccctgctggatggttag MLLLVTSLLLCELPHPAFLLIPQVQLQQSGAELAKPGASVKLSCKASGYTFTSYWMHWV KQRPGQGLEW1GY1KPSSGYTKYNQKFKDKATLTADKSSSTAYMQLSSLTYEDSAVYYC ARDYYGSSSWFAYWGQGTLVTVSAGGGGSGGGGSDIVMTQSQRFMSTSVGDRVSVTCK ASQNVGTNVAWYQQKPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLA EYFCQQYNSYPYMYTFGGGTKLELNSLQVLDYSLNHDMTSKKQELQHFPSSLAFLNLTQ NDFACTCEHQSFLQWIKDQRQLLVEVERMECATPSDKQGMPVLSLNITCQMNKTIIGVSV LSVLWSWAVLVYKFYFHLMLLAGCIKYGRGENIYDAFVIYSSQDEDWVRNELVKNLE EGVPPFQLCLHYRDFIPGVAIAANIIHEGFHKSRKVIVWSQHFIQSRWCIFEYEIAQTWQF LSSRAGIIFIVLQKVEKTLLRQQVELYRLLSRNTYLEWEDSVLGRHIFWRRLRK ALLDG ATGCTCTTGTTGGTGACGAGTCTCCTGCTGTGTGAACTGCCGCACCCAGCATTTCTTTT GATTCCGCAGGTTCAGCTGCAGCAGTCTGGTGCCGAACTGGCTAAACCTGGCGCCTC TGTGAAGCTGAGCTGTAAAGCCAGCGGCTACACCTTCACCAGCTACTGGATGCACTG GGTCAAGCAGAGGCCTGGACAGGGACTCGAGTGGATCGGCTACATCAAGCCTAGCA GCGGCTATACCAAGTACAACCAGAAGTTCAAGGACAAGGCCACACTGACCGCCGAC AAGAGCAGCAGCACAGCCTACATGCAGCTGAGCAGCCTGACCTACGAGGACAGCGC CGTGTACTACTGCGCCAGAGATTACTACGGCAGCAGCAGTTGGTTCGCCTATTGGGG CCAGGGCACACTGGTTACAGTTTCTGCTGGTGGCGGAGGATCTGGCGGAGGCGGATC TGATATCGTGATGACCCAGAGCCAGCGGTTCATGAGCACAAGCGTGGGCGATAGAGT GTCCGTGACATGCAAGGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCA GAAGCCTGGGCAGTCTCCCAAGGCTCTGATCTACAGCGCCAGCTACAGATACAGCGG CGTGCCCGATAGATTCACAGGCAGCGGCTCTGGCACCGACTTCACCCTGACAATCAG CAACGTGCAGAGCGAGGACCTGGCCGAGTACTTCTGCCAGCAGTACAACAGCTACCC CTACATGTACACCTTCGGCGGAGGCACCAAGCTGGAACTGAACTCCCTCCAGGTTCTT GATTACAGTCTCAATCACATAATGACTTCCAAAAAACAGGAACTACAGCATTTTCCA AGTAGTCTAGCTTTCTTAAATCTTACTCAGAATGACTTTGCTTGTACTTGTGAACACC AGAGTTTCCTGCAATGGATCAAGGACCAGAGGCAGCTCTTGGTGGAAGTTGAACGAA TGGAATGTGCAACACCTTCAGATAAGCAGGGCATGCCTGTGCTGAGTTTGAATATCA CCTGTCAGATGAATAAGACCATCATTggtgtgtcggtcctcagtgtgctgtagtatctgttgtagcagtctggtctata agttctattttcacctgatgcttcttgctggctgcataaagtatggtagaggtgaaaacatctatgatgcctttgttatctactcaagccaggatgag gactgggtaaggaatgagctagtaaagaatttagaagaaggggtgcctccatttcagctctgcctcactacagagactttattcccggtgtgg

[0584]

[0585] ccattgctgccaacatcatccatgaaggttccataaaagccgaaaggtgattgtgtggtgtcccagcactcatccagagccgctggtgtatctttgaatatgagattgctcagacctggcagtttctgagcagtcgtgctggtatcatcttcatgtcctgcagaaggtggagaagaccctgctcag gcagcaggtggagctgtaccgccttctcagcaggaacacttacctggagtgggaggacagtgtcctggggcggcacatcttctggagacga ctcagaaaagccctgctggatggttag MALPVTALLLPLALLLHAARPQVQLQQSGAELAKPGASVKLSCKASGYTFTSYWMHWV KQRPGQGLEWIGYKPSSGYTKYNQKFKDKATLTADKSSSTAYMQLSSLTYEDSAVYYC ARDYYGSSSWFAYWGQGTLVTVSAGGGGSGGGGSDIVMTQSQRFMSTSVGDRVSVTCK ASQNVGTNVAWYQQKPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLA EYFCQQYNSYPYMYTFGGGTKLEALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIAS QPLSLRPEASRPAAGGAVHTRGLDVDASSLWIPVVAALLLFLVLSLVFILVLKYRGLIKY WFHTPPSIPLQIEEYLKDPTQPILEALDKDSSPKDDVWDSVSIISFPEKEQEDVLQTLGGGG SGGGGSGGGGSCFYTKKTNPLKEKSITLPKSLISWRSATLETKPESKYVSLTTSYQPFSLEKE WCEEPLSPATVPGMHTEDNPGKVEHTEELSSITEWTTEENIPDWPGSHLTPIERESSSP LSSNQSEPGSIALNSYHSRNCSESDHSRNGFDTDSSCLESHSSLSDSEFPPNNKGEIKTEGQ ELITVIKAPTSFGYDKPHVLVDLLVDDSGKESLIGYRPTEDSKEFS ATGGCTCTGCCTGTTACAGCTCTGCTGCTGCCTCTGGCTCTGCTTCTGCATGCTGCTAG ACCTCAGGTTCAGCTGCAGCAGTCTGGTGCCGAACTGGCTAAACCTGGCGCCTCTGT GAAGCTGAGCTGTAAAGCCAGCGGCTACACCTTCACCAGCTACTGGATGCACTGGGT CAAGCAGAGGCCTGGACAGGGACTCGAGTGGATCGGCTACATCAAGCCTAGCAGCG GCTATACCAAGTACAACCAGAAGTTCAAGGACAAGGCCACACTGACCGCCGACAAG AGCAGCAGCACAGCCTACATGCAGCTGAGCAGCCTGACCTACGAGGACAGCGCCGT GTACTACTGCGCCAGAGATTACTACGGCAGCAGCAGTTGGTTCGCCTATTGGGGCCA GGGCACACTGGTTACAGTTTCTGCTGGTGGCGGAGGATCTGGCGGAGGCGGATCTGA TATCGTGATGACCCAGAGCCAGCGGTTCATGAGCACAAGCGTGGGCGATAGAGTGTC CGTGACATGCAAGGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGA AGCCTGGGCAGTCTCCCAAGGCTCTGATCTACAGCGCCAGCTACAGATACAGCGGCG TGCCCGATAGATTCACAGGCAGCGGCTCTGGCACCGACTTCACCCTGACAATCAGCA ACGTGCAGAGCGAGGACCTGGCCGAGTACTTCTGCCAGCAGTACAACAGCTACCCCT ACATGTACACCTTCGGCGGAGGCACCAAGCTGGAAGCCCTGAGCAACAGCATCATGT ACTTCAGCCACTTCGTGCCCGTGTTTCTGCCCGCCAAGCCTACAACAACCCCTGCTCC TAGACCACCTACACCAGCTCCTACAATCGCCAGCCAGCCTCTGTCTCTGAGGCCTGAA GCTTCTAGACCAGCTGCTGGCGGAGCCGTGCATACCAGAGGACTTGATGTCGACGCT AGCtctcttggattccagttgttgctgctttactactctttctagtgcttagcctggtattcatcctggtcctgaaatatagaggcctgattaaatac tggttcacactccaccaagcatcccattacagatagaagagtatttaaaagacccaactcagcccatctagaggccttggacaaggacagct caccaaaggatgacgtctgggactctgtgtccattatctcgtttccggaaaaggagcaagaagatgttctccaaacgcttGGTGGCGG AGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGATCTtgtttttatattaagaaaattaatccattgaagga aaaaagcataatattacccaagtccttgatctctgtggtaagaagtgctactttagagacaaaacctgaatcaaaatatgtatcactcatcacgtc ataccagccattttccttagaaaaggaggtggtctgtgaagagccgttgtctccagcaacagttccaggcatgcataccgaagacaatccagg aaaagtggaacatacagaagaactttctagtataacagaagtggtgactactgaagaaaatattcctgacgtggtcccgggcagccatctgac tccaatagagagagagagttcttcacctttaagtagtaaccagtctgaacctggcagcatcgctttaaactcgtatcactccagaaatgttctga gagtgatcactccagaaatggttttgatactgattccagctgtctggaatcacatagctccttatctgactcagaatttcccccaaataataaaggt gaaataaaaacagaaggacaagagctcataaccgtaataaaagcccccacctcctttggttatgataaaccacatgtgctagtggatctacttg tggatgatagcggtaaagagtccttgattggttatagaccaacagaagattccaaagaattttcatga MALPVTALLLPLALLLHAARPQVQLQQSGAELAKPGASVKLSCKASGYTFTSYWMHWV KQRPGQGLEWIGYIKPSSGYTKYNQKFKDKATLTADKSSSTAYMQLSSLTYEDSAVYYC ARDYYGSSSWFAYWGQGTLVTVSAGGGGSGGGGSDIVMTQSQRFMSTSVGDRVSVTCK ASQNVGTNVAWYQQKPGQSPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISNVQSEDLA EYFCQQYNSYPYMYTFGGGTKLEALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIAS QPLSLRPEASRPAAGGAVHTRGLDVDASSLWIPVVAALLLFLVLSLVFICFYIKKINPLKE KSIILPKSLISWRSATLETKPESKYVSLITSYQPFSLEKEWCEEPLSPATVPGMHTEDNPG KVEHTEELSSITEVVTTEENIPDWPGSHLTPIERESSSPLSSNQSEPGSIALNSYHSRNCSES DHSRNGFDTDSSCLESHSSLSDSEFPPNNKGEIKTEGQELITVIKAPTSFGYDKPHVLVDLL VDDSGKESLIGYRPTEDSKEFSGGGGSGGGGSGGGGSGGGGSLVLKYRGLIKYWFHTPPS IPLQIEEYLKDPTQPILEALDKDSSPKDDVWDSVSIISFPEKEQEDVLQTL ATGGCTCTGCCTGTTACAGCTCTGCTGCTGCCTCTGGCTCTGCTTCTGCATGCTGCTAG ACCTCAGGTTCAGCTGCAGCAGTCTGGTGCCGAACTGGCTAAACCTGGCGCCTCTGT GAAGCTGAGCTGTAAAGCCAGCGGCTACACCTTCACCAGCTACTGGATGCACTGGGT CAAGCAGAGGCCTGGACAGGGACTCGAGTGGATCGGCTACATCAAGCCTAGCAGCG

[0586]

[0587] GCTATACCAAGTACAACCAGAAGTTCAAGGACAAGGCCACACTGACCGCCGACAAGAGCAGCAGCACAGCCTACATGCAGCTGAGCAGCCTGACCTACGAGGACAGCGCCGT GTACTACTGCGCCAGAGATTACTACGGCAGCAGCAGTTGGTTCGCCTATTGGGGCCA GGGCACACTGGTTACAGTTTCTGCTGGTGGCGGAGGATCTGGCGGAGGCGGATCTGA TATCGTGATGACCCAGAGCCAGCGGTTCATGAGCACAAGCGTGGGCGATAGAGTGTC CGTGACATGCAAGGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGA AGCCTGGGCAGTCTCCCAAGGCTCTGATCTACAGCGCCAGCTACAGATACAGCGGCG TGCCCGATAGATTCACAGGCAGCGGCTCTGGCACCGACTTCACCCTGACAATCAGCA ACGTGCAGAGCGAGGACCTGGCCGAGTACTTCTGCCAGCAGTACAACAGCTACCCCT ACATGTACACCTTCGGCGGAGGCACCAAGCTGGAAGCCCTGAGCAACAGCATCATGT ACTTCAGCCACTTCGTGCCCGTGTTTCTGCCCGCCAAGCCTACAACAACCCCTGCTCC TAGACCACCTACACCAGCTCCTACAATCGCCAGCCAGCCTCTGTCTCTGAGGCCTGAA GCTTCTAGACCAGCTGCTGGCGGAGCCGTGCATACCAGAGGACTTGATGTCGACGCT AGCtctctttggattccagttgttgctgctttactactctttctagtgcttagcctggtattcatctgtttttatattaagaaaattaatccattgaagga aaaaagcataatattacccaagtccttgatctctgtggtaagaagtgctactttagagacaaaacctgaatcaaaatatgtatcactcatcacgtc ataccagccatttccttagaaaaggaggtggtctgtgaagagccgttgtctccagcaacagttccaggcatgcataccgaagacaatccagg aaaagtggaacatacagaagaactttctagtataacagaagtggtgactactgaagaaaatattcctgacgtggtcccgggcagccatctgac tccaatagagagagagagttcttcacctttaagtagtaaccagtctgaacctggcagcatcgctttaaactcgtatcactccagaaattgttctga gagtgatcactccagaaatggttttgatactgattccagctgtctggaatcacatagctccttatctgactcagaatttcccccaaataataaaggt gaaataaaaacagaaggacaagagctcataaccgtaataaaagcccccacctcctttggttatgataaaccacatgtgctagtggatctacttg tggatgatagcggtaaagagtccttgattggttatagaccaacagaagattccaaagaattttcaGGTGGCGGAGGATCTGGT GGCGGAGGATCTGGTGGCGGAGGATCTGGTGGCGGAGGATCTctggtcctgaaatatagaggcctg attaaatactggtttcacactccaccaagcatcccattacagatagaagagtattaaaagacccaactcagcccatcttagaggccttggacaa

[0588]

[0589] ggacagctcaccaaaggatgacgtctgggactctgtgtccattatctcgtttccggaaaaggagcaagaagatgttctccaaacgctttga

[0590] EQUIVALENTS

[0591] Various aspects of the present disclosure may be used alone, in combination, or in a variety of arrangements not specifically discussed in the embodiments described in the foregoing and is, therefore, not limited in its application to the details and arrangement of components set forth in the foregoing description or illustrated in the drawings. For example, aspects described in one embodiment may be combined in any manner with aspects described in other embodiments.

Claims

CLAIMSWhat is claimed:

1. A macrophage chimeric antigen receptor (CAR) of the formula:Formula I . R1 - R2 - R3 - R4,Formula II. R5 - R2 - R6 - R7,Formula III. R1 - R8 - R3 - R4,Formula IV. R5 - R8 - R6 - R7,Formula V. R9 - R2 - R6 - R7,Formula VI. R9 - R8 - R6 - R7,Formula VII. R1 -R10 -R11 -R12,Formula VIII. R5 - R13 - R14 - R15,Formula IX. R1 - R16 - R17 - R12, orFormula X. R9 - R18 - R19 - R15;wherein:R1 comprises an extracellular binding domain;R2 is a truncated hinge or spacer region derived from tolldike receptor 4 (TLR4);R3 is a transmembrane domain derived from tolldike receptor 4 (TLR4);R4 comprises an intracellular signaling domain derived from toll-like receptor 4 (TLR4); R5 comprises an extracellular domain means for binding HER2;R6 is a protein means derived from toll-like receptor 4 (TLR4);R7 is an intracellular means derived from toll-like receptor 4 (TLR4);R8 is a short truncated hinge or spacer region derived from toll-like receptor 4 (TLR4); R9 comprises an extracellular domain means for binding ALK;RIO is a hinge or spacer region derived from cluster of differentiation 28 (CD28);R11 is a transmembrane domain derived from cluster of differentiation 28 (CD28);R12 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2);R13 is a protein means derived from cluster of differentiation 28 (CD28) for linking an extracellular means for binding HER2 and a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane;R14 is a protein means derived from cluster of differentiation 28 (CD28) for traversing a cell membrane;R15 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl) and from interferon-gamma receptor 2 (IFNgR2);R16 is a hinge or spacer region derived from cluster of differentiation 8 (CD8);R17 is a transmembrane domain derived from interferon-gamma receptor 1 (IFNgRl);R18 is a protein means derived from cluster of differentiation 8 (CD8); andR19 is a protein means derived from interferon-gamma receptor 1 (IFNgRl).

2. The macrophage CAR of claim 1, wherein R1 is of the formula:Formula XI R20 - R21wherein:R20 is a signal peptide derived from immunoglobulin K (IgK); andR21 is an antibody fragment;Formula XIII. R22 -R21wherein:R22 is a signal peptide derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR); andR21 is an antibody fragment; orFormula XV R27 - R21wherein:R27 is a signal peptide derived from cluster of differentiation 8 (CD8); and R21 is an antibody fragment.

3. The macrophage CAR of claim 1, wherein R5 is of the formula:Formula XII. R23 - R25wherein:R23 is a protein means derived from immunoglobulin K (IgK); andR25 an antibody fragment means for binding HER2.

4. The macrophage CAR of claim 1, wherein R9 is of the formula:Formula XIV. R24 - R26wherein:R24 is a protein means derived from granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR); andR26 an antibody fragment means for binding ALK; orFormula XVI. R28 - R26wherein:R28 is a protein means derived from cluster of differentiation 8 (CD8); and R26 an antibody fragment means for binding ALK.

5. The macrophage CAR of claim 1, wherein R12 is of the formula:Formula XVII. R29 - R30wherein:R29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl); andR30 comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2); orFormula XVIII. R30 - R29wherein:R30 comprises an intracellular signaling domain derived from interferon-gamma receptor 2 (IFNgR2); andR29 comprises an intracellular signaling domain derived from interferon-gamma receptor 1 (IFNgRl).

6. The macrophage CAR of any one of claims 1, 3, or 4, wherein R15 is of the formula:Formula XIX. R31 - R32wherein:R31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl); andR32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2); orFormula XX. R32 - R31wherein:R32 comprises a protein means derived from interferon-gamma receptor 2 (IFNgR2); andR31 comprises a protein means derived from interferon-gamma receptor 1 (IFNgRl).

7. The macrophage CAR of claim 1, wherein:R1 comprises, consists essentially of, or consists of SEQ ID NO: 1, SEQ ID NO: 3, or SEQ ID NO: 5;R5 comprises, consists essentially of, or consists of SEQ ID NO: 1; orR9 comprises, consists essentially of, or consists of SEQ ID NO: 3 or SEQ ID NO: 5.

8. The macrophage CAR of claim 1, wherein:R2 comprises, consists essentially of, or consists of SEQ ID NO: 7;R8 comprises, consists essentially of, or consists of SEQ ID NO: 13;RIO comprises, consists essentially of, or consists of SEQ ID NO: 15;R13 comprises, consists essentially of, or consists of SEQ ID NO: 15;R16 comprises, consists essentially of, or consists of SEQ ID NO: 23; orR18 comprises, consists essentially of, or consists of SEQ ID NO: 23.

9. The macrophage CAR of claim 1, wherein:R3 comprises, consists essentially of, or consists of SEQ ID NO: 9;R6 comprises, consists essentially of, or consists of SEQ ID NO: 9;R11 comprises, consists essentially of, or consists of SEQ ID NO: 17;R14 comprises, consists essentially of, or consists of SEQ ID NO: 17;R17 comprises, consists essentially of, or consists of SEQ ID NO: 25; orR19 comprises, consists essentially of, or consists of SEQ ID NO: 25.

10. The macrophage CAR of claim 1, wherein:R4 comprises, consists essentially of, or consists of SEQ ID NO: 11;R7 comprises, consists essentially of, or consists of SEQ ID NO: 11;R12 comprises, consists essentially of, or consists of SEQ ID NO: 19 or SEQ ID NO: 21; R15 comprises, consists essentially of, or consists of SEQ ID NO: 19 or SEQ ID NO: 21.

11. The macrophage CAR of claim 1, wherein the macrophage CAR is of the formula:Formula I. R1 - R2 - R3 - R4, orFormula III. R1 - R8 - R3 - R4.

12. The macrophage CAR of claim 1, wherein the macrophage CAR is of the formula:Formula VII. R1 - RIO - R11 - R12, orFormula IX. R1 - R16 - R17 - R12.

13. A macrophage CAR comprising SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, or SEQ ID NO: 63.

14. A vector for engineering a macrophage, wherein the vector is a retroviral vector, comprising a nucleic acid encoding the macrophage CAR of any one of claims 1-13.

15. An engineered macrophage, comprising the macrophage CAR of any one of claims 1-13.

16. A pharmaceutical composition, comprising:the engineered macrophage of claim 15; anda pharmaceutically acceptable carrier.

17. A method of treating cancer, comprising administering to a subject with cancer:the engineered macrophage of claim 15; orthe pharmaceutical composition of claim 16.

18. Use of the engineered macrophage of claim 15, in the manufacture of a medicament for treating cancer.

19. The use of claim 18, wherein the cancer is ALK-positive cancer, optionally wherein the cancer is selected from the list consisting of: neuroblastoma, non-small cell lung cancer, anaplastic large cell lymphoma, inflammatory myofibroblastic tumor, diffuse large B-cell lymphoma, undifferentiated anaplastic thyroid carcinoma, breast cancer, colorectal cancer, squamous cell carcinoma, rhabdomyosarcoma, renal cell carcinoma.

20. The use of claim 18, wherein the cancer is HER2-positive cancer, optionally wherein the cancer is selected from the list consisting of: breast cancer, bladder cancer, pancreatic cancer, ovarian cancer, stomach cancer, colorectal cancer, non-small cell lung cancer, head and neck cancer, cholangiocarcinoma, prostate cancer, cervical cancer.

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