Chimeric antigen receptor with modified hinge region and uses thereof
The present disclosure provides a chimeric antigen receptor (CAR) molecule comprising a modified hinge domain, wherein the hinge domain comprises the amino acid sequence of SEQ ID NO:1. Cells expressing a CAR comprising the modified hinge domain disclosed herein are shown 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 domain, a transmembrane domain, a costimulatory domain, and an intracellular signaling domain, wherein the hinge domain comprising the amino acid sequence of SEQ ID NO: 1.
2 . The isolated CAR of claim 1 , wherein the hinge domain is encoded by a nucleotide sequence comprising the sequence of SEQ ID NO:2.
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 binds to CD19 and comprises a light chain complementary determining region 1 (LC CDR1) having the amino acid sequence of SEQ ID NO:3, a light chain complementary determining region 2 (LC CDR2) having the amino acid sequence of SEQ ID NO:4, a light chain complementary determining region 3 (LC CDR3) having the amino acid sequence of SEQ ID NO:5, and a heavy chain complementary determining region 1 (HC CDR1) having the amino acid sequence of SEQ ID NO:6, a heavy chain complementary determining region 2 (HC CDR2) having the amino acid sequence of SEQ ID NO: 7, and a heavy chain complementary determining region 3 (HC CDR3) having the amino acid sequence of SEQ ID NO: 8.
6 . The isolated CAR of claim 1 , wherein the antigen binding domain comprises a scFv that binds to CD19, said scFv comprises the amino acid sequence of SEQ ID NO:11.
7 . The isolated CAR of claim 1 , wherein the transmembrane domain comprises a transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of T cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154.
8 . 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).
9 . 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.
10 . A nucleic acid construct comprising one or more nucleic acid sequences, said nucleic acid sequences encode the isolated CAR of claim 1 .
11 . An expression vector comprising the nucleic acid construct of claim 10 .
12 . A cell comprising the expression vector of claim 11 .
13 . The cell of claim 12 , wherein the cell is an immune cell.
14 . The cell of claim 13 , wherein the immune cell is a T cell.
15 . A composition comprising the cell of claim 12 and a pharmaceutically acceptable carrier.