CHIMERIC ANTIGEN RECEPTOR WITH MODIFIED HINGE AND TRANSMEMBRANE DOMAIN AND USES THEREOF
The present disclosure provides a chimeric antigen receptor (CAR) molecule comprising a modified hinge and transmembrane domain (HTM-1), wherein the modified hinge and transmembrane domain comprises the amino acid sequence of SEQ ID NO:2. Cells expressing CAR with the modified HTM-1 domain disclosed herein are expected to have enhanced anti-tumor activities with reduced release of pro-inflammatory cytokines.
1 . An isolated chimeric antigen receptor (CAR) molecule comprising an antigen binding domain, a hinge and transmembrane domain, a costimulatory domain, and an intracellular signaling domain, wherein the hinge and transmembrane domain comprises the amino acid sequence of SEQ ID NO:2.
2 . The isolated CAR of claim 1 , wherein the hinge and transmembrane domain is encoded by a nucleotide sequence comprising the sequence of SEQ ID NO:1.
3 . The isolated CAR of claim 1 , wherein the antigen binding domain is a single chain antibody or single chain antibody fragment.
4 . The isolated CAR of claim 1 , wherein the antigen binding domain binds to a target antigen selected from the group consisting of CD19, CD20, CD22, CD33, CD123, BCMA, CLL1, CD7, CS1, CEA, AFP, PSMA, GPC3, GD2, EGFRVIII, NKG2D, Mesothelin, Claudin 18.2, ROR3, and Muc1.
5 . The isolated CAR of claim 1 , wherein the antigen binding domain comprises a scFv that binds to CD19, said scFv comprises the amino acid sequences of SEQ ID NOs:4 and 6.
6 . The isolated CAR of claim 1 , wherein the costimulatory domain comprises a functional signaling domain of a protein selected from the group consisting of OX40, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), and 4-1BB (CD137).
7 . The isolated CAR of claim 1 , wherein the intracellular signaling domain comprises an intracellular signaling domain of CD3 zeta, or FcR gamma, or a functional fragment thereof.
8 . A nucleic acid construct comprising one or more nucleic acid sequences, said nucleic acid sequences encode the isolated CAR of claim 1 , or fragments thereof.
9 . An expression vector comprising the nucleic acid construct of claim 8 .
10 . A cell comprising the expression vector of claim 9 .
11 . The cell of claim 10 , wherein the cell is an immune cell.
12 . The cell of claim 11 , wherein the immune cell is a T cell.
13 . A composition comprising the cell of claim 10 and a pharmaceutically acceptable carrier.