Methods and compositions for cell-surface co-localization of chimeric proteins
AssigneeSenti Biosciences, Inc.
InventorsAlba Gonzalez Junca, Nicholas Frankel, Maelig Goulwen Morvan
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Abstract
The present disclosure provides compositions and methods related to the co-localization of chimeric proteins on a cell surface. In particular, the present disclosure provides methods and compositions for facilitating the cell-surface co-localization of chimeric polypeptides using dimerization domains to promote their functional interaction.
Claims19
A chimeric protein system comprising:
- a first polypeptide comprising a first antigen binding domain, a first transmembrane domain, and a first dimerization domain;
- and a second polypeptide comprising a second antigen binding domain, a second transmembrane domain, and a second dimerization domain;
- wherein the first dimerization domain is capable of binding to the second dimerization domain, optionally wherein the first dimerization domain or the second dimerization domain is selected from the group consisting of CD94, NKG2A, and NKG2C.
The chimeric protein system of claim 1 or 2, wherein the first dimerization domain comprises a dimerization domain from CD94, and the second dimerization domain comprises a dimerization domain from NKG2A or a dimerization domain from NKG2C, optionally wherein the second dimerization domain comprises a sequence selected from the group consisting of SEQ ID NO:
- 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, and 90.
The chimeric protein system of any one of claims 1 to 6, wherein the first polypeptide is a chimeric antigen receptor, optionally wherein the first polypeptide is an activating chimeric antigen receptor or an inhibitory chimeric antigen receptor.
The chimeric protein system of any one of claims 1 to 9, wherein the second polypeptide is a chimeric antigen receptor, optionally wherein the second polypeptide is an activating chimeric antigen receptor or an inhibitory chimeric antigen receptor.
The chimeric protein system of any one of claims 1 to 12, wherein the antigen binding domain comprises a F(ab) fragment, a F(ab′) fragment, a single chain variable fragment (scFv), a single domain antibody, a diabody, a VHH fragment, or a synthetic epitope.
The chimeric protein system of claim 13, wherein the antigen binding domain binds an antigen expressed on a cancer cell, optionally wherein the cancer comprises glioblastoma, neuroblastoma, breast cancer, colorectal cancer, prostate cancer, bladder cancer, liver cancer, lung cancer, pancreatic cancer, ovarian cancer, gastric cancer, endometrial cancer, cervical cancer, leukemia, lymphoma, or myeloma, optionally wherein the antigen binding domain is specific for carcinoembryonic antigen (CEA), Mesothelin, Ax1, GPC3, FLT3, CD33, TROP2, MUC1, MUC16, IL13Ra, ErbB2 (HER2/neu), epithelial cell adhesion molecule (EpCAM), epidermal growth factor receptor (EGFR), EGFR variant III (EGFRvIII), CD19, CD20, CD30, CD40, disialoganglioside GD2, ductal-epithelial mucine, gp36, TAG-72, glycosphingolipids, glioma-associated antigen, alphafetoprotein (AFP), lectin-reactive AFP, thyroglobulin, RAGE 1, MN-CA IX, human telomerase reverse transcriptase, RU1, RU2 (AS), intestinal carboxyl esterase, mut hsp70-2, M-CSF, prostate specific antigen (PSA), PAP, NY-ESO-1, LAGA-1a, p53, prostein, PSMA, prostate-carcinoma tumor antigen-1 (PCTA-1), MAGE, ELF2M, neutrophil elastase, ephrin B2, CD22, insulin growth factor (IGF)-1, IGF-11, IGF-1 receptor, NKG2D, BCMA (CD269, TNFRSF 17), Claudin18.2, B7-H3, or Ror1.
The chimeric protein system of any one of claims 1 to 16, wherein the transmembrane domain is selected from the group consisting of:
- a LAX transmembrane domain, a CD25 transmembrane domain, a CD7 transmembrane domain, a LAT transmembrane domain, a transmembrane domain from a LAT mutant, a BTLA transmembrane domain, a CDS transmembrane domain, a CD28 transmembrane domain, a CD3zeta transmembrane domain, a CD4 transmembrane domain, a 4-IBB transmembrane domain, an OX40 transmembrane domain, an ICOS transmembrane domain, a 2B4 transmembrane domain, a PD-1 transmembrane domain, a CTLA4 transmembrane domain, a BTLA transmembrane domain, a TIM3 transmembrane domain, a LIR1 transmembrane domain, an NKG2A transmembrane domain, a TIGIT transmembrane domain, and a LAG3 transmembrane domain, a LAIR1 transmembrane domain, a GRB-2 transmembrane domain, a Dok-1 transmembrane domain, a Dok-2 transmembrane domain, a SLAP1 transmembrane domain, a SLAP2 transmembrane domain, a CD200R transmembrane domain, an SIRPa transmembrane domain, an HAVR transmembrane domain, a GITR transmembrane domain, a PD-L1 transmembrane domain, a KIR2DL1 transmembrane domain, a KIR2DL2 transmembrane domain, a KIR2DL3 transmembrane domain, a KIR3DL1 transmembrane domain, a KIR3DL2 transmembrane domain, a CD94 transmembrane domain, a KLRG-1 transmembrane domain, a PAG transmembrane domain, a CD45 transmembrane domain, and a CEACAM1 transmembrane domain.
The chimeric protein system of any one of claims 1 to 17, wherein the first polypeptide and/or the second polypeptide further comprises one or more intracellular signaling domains, optionally wherein the one or more intracellular signaling domains are selected from the group consisting of:
- a CD3zeta-chain intracellular signaling domain, a CD97 intracellular signaling domain, a CD11a-CD18 intracellular signaling domain, a CD2 intracellular signaling domain, an ICOS intracellular signaling domain, a CD27 intracellular signaling domain, a CD154 intracellular signaling domain, a CDS intracellular signaling domain, an OX40 intracellular signaling domain, a 4-1BB intracellular signaling domain, a CD28 intracellular signaling domain, a ZAP40 intracellular signaling domain, a CD30 intracellular signaling domain, a GITR intracellular signaling domain, an HVEM intracellular signaling domain, a DAP10 intracellular signaling domain, a DAP12 intracellular signaling domain, a MyD88 intracellular signaling domain, a 2B4 intracellular signaling domain, a CD16a intracellular signaling domain, a DNAM-1 intracellular signaling domain, a KIR2DS1 intracellular signaling domain, a KIR3DS1 intracellular signaling domain, a NKp44 intracellular signaling domain, a NKp46 intracellular signaling domain, a FceR1g intracellular signaling domain, a NKG2D intracellular signaling domain, and an EAT-2 intracellular signaling domain, optionally wherein the one or more intracellular signaling domains comprises a co-stimulatory domain selected from the group consisting of:
- a CD97 intracellular signaling domain, a CD11a-CD18 intracellular signaling domain, a CD2 intracellular signaling domain, an ICOS intracellular signaling domain, a CD27 intracellular signaling domain, a CD154 intracellular signaling domain, a CDS intracellular signaling domain, an OX40 intracellular signaling domain, a 4-1BB intracellular signaling domain, a CD28 intracellular signaling domain, a ZAP40 intracellular signaling domain, a CD30 intracellular signaling domain, a GITR intracellular signaling domain, an HVEM intracellular signaling domain, a DAP10 intracellular signaling domain, a DAP12 intracellular signaling domain, a MyD88 intracellular signaling domain, a 2B4 intracellular signaling domain, a CD16a intracellular signaling domain, a DNAM-1 intracellular signaling domain, a KIR2DS1 intracellular signaling domain, a KIR3DS1 intracellular signaling domain, a NKp44 intracellular signaling domain, a NKp46 intracellular signaling domain, a FceR1g intracellular signaling domain, a NKG2D intracellular signaling domain, and an EAT-2 intracellular signaling domain.
The chimeric protein system of any one of claims 1 to 20, wherein the first polypeptide and/or the second polypeptide comprises a hinge domain located between the antigen binding domain and the transmembrane domain.
The chimeric protein system of any one of claims 1 to 21, wherein the first polypeptide and/or the second polypeptide comprises one or more linkers, optionally wherein the one or more linkers comprise a GSG linker, a Whitlow linker, an eGK linker, or any derivative thereof.
An engineered polynucleotide encoding the first polynucleotide and/or the second polynucleotide of any one of claims 1 to 23.
An expression vector comprising the engineered polynucleotide of claim 24.
An engineered cell comprising the engineered polynucleotide of claim 25, the vector of claim 25, or the first and/or second polypeptides of any one of claims 1 to 24.
The engineered cell of claim 26, wherein the cell is selected from the group consisting of:
- a T cell, a CDS+ T cell, a CD4+ T cell, a gamma-delta T cell, a cytotoxic T lymphocyte (CTL), a regulatory T cell, a viral-specific T cell, a Natural Killer T (NKT) cell, a Natural Killer (NK) cell, a B cell, a tumor-infiltrating lymphocyte (TIL), an innate lymphoid cell, a mast cell, an eosinophil, a basophil, a neutrophil, a myeloid cell, a macrophage, a monocyte, a dendritic cell, an ESC-derived cell, and an iPSC-derived cell, optionally wherein the cell is engineered to express an effector molecule, optionally wherein the engineered cell of claim 28, wherein the cell is a Natural Killer (NK) cell, optionally wherein the cell is allogeneic or autologous.
A pharmaceutical composition comprising the engineered cell of any one of claims 26 to 31, and a pharmaceutically acceptable carrier and/or a pharmaceutically acceptable excipient.
A method of treating a subject having cancer, the method comprising administering to the subject a therapeutically effective dose of the composition of claim 32, or a cell of any one of claims 26 to 31 to the subject, optionally wherein the cancer comprises glioblastoma, neuroblastoma, breast cancer, colorectal cancer, prostate cancer, bladder cancer, liver cancer, lung cancer, pancreatic cancer, ovarian cancer, gastric cancer, endometrial cancer, cervical cancer, leukemia, lymphoma, or myeloma.
A method of enhancing immune cell-mediated killing of a cancer cell in a subject in need thereof, the method comprising administering a therapeutically effective dose of the composition of claim 32, or a cell of any one of claims 26 to 31, to the subject, optionally wherein the cancer comprises a solid tumor, optionally wherein the cancer comprises glioblastoma, neuroblastoma, breast cancer, colorectal cancer, prostate cancer, bladder cancer, liver cancer, lung cancer, pancreatic cancer, ovarian cancer, gastric cancer, endometrial cancer, cervical cancer, leukemia, lymphoma, or myeloma.
A method of reducing off-target killing of a healthy cell in a subject, the method comprising administering a therapeutically effective dose of the composition of claim 32, or a cell of any one of claims 26 to 31, to the subject, wherein the subject has been diagnosed with cancer, optionally wherein the cancer comprises a solid tumor, optionally wherein the cancer comprises glioblastoma, neuroblastoma, breast cancer, colorectal cancer, prostate cancer, bladder cancer, liver cancer, lung cancer, pancreatic cancer, ovarian cancer, gastric cancer, endometrial cancer, cervical cancer, leukemia, lymphoma, or myeloma.
Dependencies are inferred from the claim text.