Compound chimeric antigen receptor (cCAR) targeting multiple antigens, compositions and methods of use thereof
In one embodiment, the present disclosure provides an engineered cell having a first chimeric antigen receptor polypeptide including a first antigen recognition domain, a first signal peptide, a first hinge region, a first transmembrane domain, a first costimulatory domain, and a first signaling domain; and a second chimeric antigen receptor polypeptide including a second antigen recognition domain, a second signal peptide, a second hinge region, a second transmembrane domain, a second co-stimulatory domain, and a second signaling domain; wherein the first antigen recognition domain is different than the second antigen recognition domain.
1. An ex vivo engineered T cell or NK cell co-expressing two distinct chimeric antigen receptor (CAR) units at the cell surface, wherein the engineered T cell or NK cell comprises a nucleotide sequence comprising from 5′ to 3′ a first polynucleotide encoding a first chimeric antigen receptor polypeptide (first CAR), a second polynucleotide encoding a second chimeric antigen receptor polypeptide (second CAR), a nucleotide encoding porcine teschovirus-1 2A (P2A), thoseaasigna virus 2A (T2A), FMDV 2A (F2A) or equine rhinitis A virus (ERAV) 2A (E2A1 disposed between the first CAR and second CAR, under the transcriptional control of a single promoter, wherein:
(i.) the first CAR polypeptide comprises a first antigen recognition domain; a first signal peptide; a first hinge region; a first transmembrane domain; a first co-stimulatory domain; and a first signaling domain; and
(ii.) the second CAR comprises a second antigen recognition domain; a second signal peptide; a second hinge region; a second transmembrane domain; a second co-stimulatory domain; and a second signaling domain;
wherein the first antigen recognition domain and the second antigen recognition domain are different;
wherein the engineered T cell or NK cell comprises an enhancer selected from the group consisting of IL-15/IL-15sushi, IL-15/IL-15 sushi anchor, 4-1BBL, and IL-15: and
wherein the engineered T cell or NK cell comprises SEQ ID NO: 1, SEQ ID NO: 28, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 50, SEQ ID NO: 52, or SEQ ID NO: 60.
2. The engineered cell according to claim 1 , wherein the engineered T cell or NK cell comprises SEQ ID NO: 60.
3. The engineered cell according to claim 1 , wherein the enhancer is secreted by the engineered cell.
4. The engineered cell according to claim 1 , wherein the engineered T cell is an NK T cell.
5. A method of treating a cell proliferative disease comprising administering an engineered T cell or NK cell according to claim 1 to a patient in need thereof.
6. The method according to claim 5 , wherein the cell proliferative disease comprises a t-cell malignancy, leukemia, or a lymphoma.
7. The method according to claim 5 , wherein the engineered T cell is an NK T cell.