CHIMERIC RECEPTORS WITH 4-1BB STIMULATORY SIGNALING DOMAIN
The present invention relates to a chimeric receptor capable of signaling both a primary and a co-stimulatory pathway, thus allowing activation of the co-stimulatory pathway without binding to the natural ligand. The cytoplasmic domain of the receptor contains a portion of the 4-1BB signaling domain. Embodiments of the invention relate to polynucleotides that encode the receptor, vectors and host cells encoding a chimeric receptor, particularly including T cells and natural killer (NK) cells and methods of use.
1 . A chimeric antigen receptor comprising: (a) an extracellular ligand-binding domain comprising an anti-CD 19 single chain variable fragment (scFv) domain; (b) a transmembrane domain; and (c) a cytoplasmic domain comprising a 4-1BB signaling domain and a CD3ζ signaling domain wherein the CD3ζ signaling domain comprises the amino acid sequence of SEQ ID No. 18.
2 . The chimeric antigen receptor of claim 1 , wherein the extracellular binding domain comprises the amino acid sequence of SEQ. ID. No. 10.
3 . The chimeric antigen receptor of claim 1 wherein the transmembrane domain comprises the amino acid sequence of SEQ ID No. 14.
4 . The chimeric antigen receptor of claim 1 wherein the 4-1BB signaling domain comprises the amino acid sequence of SEQ ID No. 16.
5 . The chimeric antigen receptor of claim 1 comprising the amino acid sequence of SEQ. ID. No. 6.
6 . A polynucleotide encoding the CAR of claim 1 .
7 . The polynucleotide of claim 6 , wherein the a cytoplasmic domain is encoded by the nucleic acid sequence of SEQ ID No. 15.
8 . The polynucleotide claim 6 , wherein the extracellular binding domain is encoded by the nucleic acid sequence of SEQ. ID. No. 9.
9 . The polynucleotide of claim 6 , wherein the transmembrane domain is encoded by the nucleic acid sequence of SEQ ID No. 13.
10 . The polynucleotide of claim 6 , wherein the CD3ζ signaling domain is encoded by the nucleic acid sequence of either SEQ ID No. 17.
11 . The polynucleotide of claim 6 comprising the nucleotide sequence of SEQ. ID. No. 5.
12 . A vector comprising the polynucleotide of claim 11 .
13 . The vector of claim 12 which is a retroviral vector.
14 . An isolated host cell comprising the polynucleotide of claim 11 .
15 . The isolated host cell of claim 14 which is a T lymphocyte.
16 . The isolated host cell of claim 14 which is an autologous cell.
17 . The isolated host cell of claim 14 which is isolated from a patient having a cancer of B cell origin.
18 . The isolated host cell of claim 16 , wherein the autologous cell is an autologous T lymphocyte.
19 . The isolated host cell of claim 18 , wherein the autologous T lymphocyte is derived from a blood or tumor sample of a patient having a cancer of B cell origin and activated and expanded in vitro.
20 . The isolated host cell of claim 17 which is isolated from a patient having lymphoblastic leukemia, B-lineage acute lymphoblastic leukemia, B-cell chronic lymphocytic leukemia or B-cell non-Hodgkin's lymphoma.
21 . The isolated host cell of claim 17 , which is isolated from a patient having lung cancer, melanoma, breast cancer, prostate cancer, colon cancer, renal cell carcinoma, ovarian cancer, neuroblastoma, rhabdomyosarcoma, leukemia and lymphoma, acute lymphoblastic leukemia, small cell lung cancer, Hodgkin's lymphoma, or childhood acute lymphoblastic leukemia.
22 . The isolated host cell of claim 18 , wherein the autologous T lymphocyte is derived from a blood or tumor sample of a patient having a cancer of B cell origin and activated and expanded in vitro.
23 . A method of enhancing a T lymphocyte or an NK cell activity in a mammal comprising introducing into the mammal a T lymphocyte or NK cell, which T lymphocyte or NK cell comprises a chimeric receptor comprising: (a) an extracellular ligand-binding domain comprising an anti-CD19 single chain variable fragment (scFv) domain, (b) a hinge and transmembrane domain, and (c) a cytoplasmic domain comprising a 4-1BB signaling domain and a CD3ζ signaling domain comprising the amino acid sequence of SEQ ID No. 18.
24 . A method for treating a mammal suffering from cancer comprising introducing into the mammal a T lymphocyte or an NK cell, which T lymphocyte or NK cell comprises a chimeric receptor comprising: (a) an extracellular ligand-binding domain comprising an anti-CD19 scFv domain; (b) a hinge region and transmembrane domain of CD8α; and (c) a cytoplasmic domain comprising a signaling domain of 4-1BB and a CD3ζ signaling domain comprising the amino acid sequence of SEQ ID No. 18.
25 . A method for stimulating a T cell-mediated immune response to a target cell population or tissue in a mammal comprising administering to a mammal an effective amount of a cell genetically modified to express a chimeric antigen receptor (CAR), said CAR comprising: a) an extracellular ligand-binding domain comprising an anti-CD19 single chain variable fragment (scFv) domain, (b) a hinge and transmembrane domain, and (c) a cytoplasmic domain comprising a 4-1BB signaling domain and a CD3 signaling domain comprising the amino acid sequence of SEQ ID No. 18.
26 . A method of providing an anti-tumor immunity in a mammal, the method comprising administering to the mammal an effective amount of a cell genetically modified to express a CAR, said CAR comprising: a) an extracellular ligand-binding domain comprising an anti-CD19 single chain variable fragment (scFv) domain, (b) a hinge and transmembrane domain, and (c) a cytoplasmic domain comprising a 4-1BB signaling domain and a CD3 signaling domain comprising the amino acid sequence of SEQ ID No. 18.
27 . A method of treating a mammal having a disease, disorder or condition associated with an elevated expression of a tumor antigen, the method comprising administering to the mammal an effective amount of a cell genetically modified to express a CAR, said CAR comprising comprising an antigen binding domain, a costimulatory signaling region, and a CD3 zeta signaling domain comprising the amino acid sequence of SEQ ID No. 18, thereby treating the mammal.
28 . A method of treating a human with chronic lymphocytic leukemia, the method comprising administering to the human a T cell genetically engineered to express a CAR wherein the CAR comprises an antigen binding domain, a costimulatory signaling region, and a CD3 zeta signaling domain comprising the amino acid sequence of SEQ ID No. 18.
29 . A method of generating a persisting population of genetically engineered T cells in a human diagnosed with cancer, said method comprising administering to the human a T cell genetically engineered to express a CAR, said CAR comprising: a) an extracellular ligand-binding domain comprising an anti-CD19 single chain variable fragment (scFv) domain, (b) a hinge and transmembrane domain an antigen binding domain, and (c) a cytoplasmic domain comprising a signaling region comprising a costimulatory domain and a CD3 zeta signaling domain comprising the amino acid sequence of SEQ ID No. 18.
30 . A method of expanding a population of genetically engineered T cells in a human diagnosed with cancer, the method comprising administering to the human aT cell genetically engineered to express a CAR, said CAR comprising: an antigen binding domain, a costimulatory signaling region, and a CD3 zeta signaling domain comprising the amino acid sequence of SEQ ID NO: 18, wherein the administered genetically engineered T cell produces a population of progeny T cells in the human.
31 . The method of any one of claims 24 to 28 , wherein the mammal is suffering from B-lineage acute lymphoblastic leukemia, B-cell chronic lymphocytic leukemia, B-cell non-Hodgkin's lymphoma lung cancer, melanoma, breast cancer, prostate cancer, colon cancer, renal cell carcinoma, ovarian cancer, neuroblastoma, rhabdomyosarcoma, leukemia and lymphoma, acute lymphoblastic leukemia, small cell lung cancer, Hodgkin's lymphoma, or childhood acute lymphoblastic leukemia.