Vector genetically engineered with chimeric antigen receptor and against two or more targets and application thereof
Provided are a vector genetically engineered with a chimeric antigen receptor and against two or more targets for combined treatment of disease such as human tumors, a related immune cell and application thereof. The vector genetically engineered with a chimeric antigen receptor and the related immune cell have enhanced ability to kill target cells.
1 . A nucleic acid molecule expressing at least two types of chimeric antigen receptors (CAR), comprising a nucleic sequence encoding a leader peptide, a nucleic acid sequence encoding a first antigen-binding domain, a nucleic sequence encoding a first non-antigen binding function domain, a nucleic sequence encoding a cleavable peptide, a nucleic sequence encoding a leader sequence, a nucleic acid sequence encoding a second antigen-binding domain, and a nucleic sequence encoding a first non-antigen binding function domain;
wherein said non-antigen binding function domain comprises a hinge region, a transmembrane region, a co-stimulatory domain and an intracellular signaling domain, wherein the nucleic acid molecule comprises a nucleotide sequence as set forth in SEQ ID NO: 83 or SEQ ID NO: 84.
2 . The nucleic acid molecule according to claim 1 , comprising a nucleic acid sequence encoding a promoter.
3 . The nucleic acid molecule according to claim 2 , wherein said promoter is selected from the group consisting of: EF1α, CMV, MSCV, and UbC.
4 . The nucleic acid molecule according to claim 1 , comprising a nucleic acid sequence encoding WPRE, wherein said nucleic acid sequence encoding WPRE is located downstream of said nucleic acid sequence encoding the non-antigen-binding function domain.
5 . The nucleic acid molecule according to claim 1 , comprising a nucleic acid sequence encoding a backbone vector, wherein said nucleic acid sequence encoding the backbone vector is located upstream of said nucleic acid sequence encoding the promoter.
6 . The nucleic acid molecule according to claim 5 , wherein said backbone vector comprises a viral vector.
7 . The nucleic acid molecule according to claim 5 , wherein said backbone vector comprises an HIV packaging vector.
8 . The nucleic acid molecule according to claim 1 , comprising a 5′ LTR sequence located at a 5′-terminal of said nucleic acid molecule, and a 3′ LTR sequence located at a 3′-terminal of said nucleic acid molecule.
9 . The nucleic acid molecule according to claim 1 , which expresses two types of chimeric antigen receptors.
10 . A plasmid, comprising the nucleic acid molecule of claim 1 .
11 . The plasmid according to claim 10 , comprising a viral plasmid.
12 . A cell, comprising the nucleic acid molecule of claim 1 .
13 . The cell according to claim 12 , comprising a cell selected from the group consisting of: a T cell and a NK cell.