Chimeric antigen receptors
Provided herein are therapeutic polypeptides, e.g., chimeric antigen receptors, able to direct an immune cell, e.g., a T lymphocyte to a target antigen, and able to cause the T cell to proliferate or to kill cells displaying the antigen when the antigen binds to the polypeptide, wherein the polypeptides comprise a transmembrane domain from a T cell co-inhibitory protein such as CTLA4 or PD-1. Also provided herein are T lymphocytes expressing the polypeptides, and use of such T lymphocytes to treat diseases such as cancer.
1. A polypeptide comprising (i) a transmembrane domain from PD-1, (ii) a CD3ζ intracellular signaling domain, and (iii) an extracellular domain that binds to an antigen on a tumor cell, wherein said extracellular domain is an antibody or an antigen-binding portion thereof, and wherein the intracellular domain and the extracellular domain of said polypeptide are not from PD-1, wherein said polypeptide, when expressed on the surface of a T lymphocyte, directs the T lymphocyte to kill a cell expressing said antigen.
2. The polypeptide of claim 1 , wherein said polypeptide is a chimeric antigen receptor (CAR).
3. The polypeptide of claim 1 , wherein a T lymphocyte expressing said polypeptide is stimulated to proliferate when said polypeptide binds to said antigen.
4. The polypeptide of claim 1 , wherein said antigen is a tumor-associated antigen or a tumor-specific antigen.
5. The polypeptide of claim 4 , wherein said tumor-associated antigen or tumor-specific antigen is Her2, prostate stem cell antigen (PSCA), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen-125 (CA-125), CA19-9, calretinin, MUC-1, epithelial membrane protein (EMA), epithelial tumor antigen (ETA), tyrosinase, melanoma-associated antigen (MAGE), CD34, CD45, CD99, CD117, chromogranin, cytokeratin, desmin, glial fibrillary acidic protein (GFAP), gross cystic disease fluid protein (GCDFP-15), HMB-45antigen, protein melan-A (melanoma antigen recognized by T lymphocytes; MART-1), myo-D1, muscle-specific actin (MSA), neurofilament, neuron-specific enolase (NSE), placental alkaline phosphatase, synaptophysis, thyroglobulin, thyroid transcription factor-1, the dimeric form of the pyruvate kinase isoenzyme type M2 (tumor M2-PK), CD19, CD22, CD27, CD30, CD70, GD2 (ganglioside G2), EGFRvIII (epidermal growth factor variant III), sperm protein 17 (Sp17), mesothelin, PAP (prostatic acid phosphatase), prostein, TARP (T cell receptor gamma alternate reading frame protein), Trp-p8, STEAP1 (six-transmembrane epithelial antigen of the prostate 1), an abnormal ras protein, an abnormal p53 protein, integrin av(33 (CD61), galactin, K-Ras (V-Ki-ras2 Kirsten rat sarcoma viral oncogene), or Ral-B.
6. The polypeptide of claim 1 , wherein said polypeptide additionally comprises one or more co-stimulatory domains, wherein said one or more co-stimulatory domains comprises one or more of: a co-stimulatory CD27 polypeptide sequence, a co-stimulatory CD28 polypeptide sequence, a co-stimulatory OX40 (CD134) polypeptide sequence, a co-stimulatory 4-1BB (CD137) polypeptide sequence, and a co-stimulatory inducible T-cell costimulatory (ICOS) polypeptide sequence.
7. The polypeptide of claim 1 , wherein said polypeptide comprises, in order, from N-terminus to C terminus: (i) the extracellular domain that binds to an antigen on a tumor cell; (ii) a hinge polypeptide sequence from CD28 or CTLA4; (iii) the transmembrane domain from PD-1; (iv) a costimulatory domain; and (v) the CD3ζ intracellular signaling domain.
8. The polypeptide of claim 7 , wherein said polypeptide comprises, in order, from N-terminus to C-terminus: (i) a single-chain Fv domain comprising a V L linked to V H by a flexible linker, wherein said V L and V H are from an antibody that binds said antigen; (ii) a hinge polypeptide sequence from CD28; (iii) the transmembrane domain from PD-1; (iv) a CD28 costimulatory domain; and (v) the CD3ζ intracellular signaling domain.
9. The polypeptide of claim 7 , wherein said polypeptide comprises, in order, from N-terminus to C-terminus: (i) a single-chain Fv domain comprising a V L linked to V H by a flexible linker, wherein said V L and V H are from an antibody that binds said antigen; (ii) a hinge polypeptide sequence from CTLA4; (iii) the transmembrane domain from PD-1; (iv) a CD28 costimulatory domain; and (v) the CD3ζ intracellular signaling domain.
10. A T lymphocyte comprising the polypeptide of claim 1 .