CR3022 chimeric antigen receptors and methods of use
Chimeric antigen receptors (CARs) including an antigen binding domain specifically binding to coronavirus spike protein, nucleic acids encoding the CARs, vectors including nucleic acids encoding the CARs, and immune cells expressing the CARs are provided. Methods of treating a subject with coronavirus, including administering to the subject an immune cell expressing a disclosed CAR are also provided.
1. A chimeric antigen receptor comprising at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 3 or at least 90% sequence identity to amino acids 22-740 of SEQ ID NO: 3,
wherein the chimeric antigen receptor comprises an antigen binding domain that specifically binds a coronavirus spike protein, comprising an amino acid sequence comprising variable heavy chain (VH) domain complementarity determining region 1 (CDR1), CDR2 and CDR3 amino acid sequences of amino acid positions 31-35, 50-66, and 99-108 of SEQ ID NO: 1, respectively, and variable light chain (VL) domain CDR1, CDR2 and CDR3 amino acid sequences of amino acid positions 161-177, 193-199, and 232-240 of SEQ ID NO: 1, respectively.
2. The chimeric antigen receptor of claim 1 , wherein the antigen binding domain comprises:
a VH domain that comprises the amino acid sequence of SEQ ID NO: 5;
a VL domain that comprises the amino acid sequence of SEQ ID NO: 7; or both.
3. The chimeric antigen receptor of claim 1 , wherein the antigen binding domain comprises the amino acid sequence of SEQ ID NO: 1.
4. The chimeric antigen receptor of claim 1 wherein the chimeric antigen receptor comprises the amino acid sequence of SEQ ID NO: 3 or comprises the amino acid sequence of amino acids 22-740 of SEQ ID NO: 3.
5. The chimeric antigen receptor of claim 1 , further comprising one or more additional antigen binding domains.
6. The chimeric antigen receptor of claim 5 , wherein the one or more additional antigen binding domains specifically binds a coronavirus spike protein, a coronavirus envelope protein, a coronavirus membrane protein, or any combination thereof.
7. The chimeric antigen receptor of claim 6 , wherein the antigen binding domains specifically bind a coronavirus spike protein and the antigen binding domains are different.
8. The chimeric antigen receptor of claim 1 , further comprising an inducible suicide molecule.
9. The chimeric antigen receptor of claim 8 , wherein the suicide molecule comprises caspase 9 .
10. A nucleic acid encoding the chimeric antigen receptor of claim 1 , wherein the nucleic acid comprises at least 90% sequence identity to the nucleic acid sequence of SEQ ID NO: 4 or at least 90% sequence identity to nucleotides 64-2223 of SEQ ID NO: 4.
11. The nucleic acid of claim 10 , wherein the nucleic acid comprises the nucleic acid sequence of SEQ ID NO: 4, or comprises the nucleic acid sequence of nucleotides 64-2223 of SEQ ID NO: 4.
12. A vector comprising the nucleic acid of claim 10 .
13. The vector of claim 12 , wherein the vector is a viral vector.
14. A modified immune cell expressing the chimeric antigen receptor of claim 1 .
15. The modified immune cell of claim 14 , wherein the immune cell is a T cell, natural killer (NK) cell, NKT cell, or macrophage.
16. The modified immune cell of claim 15 , wherein the immune cell is an NK cell.
17. The modified immune cell of claim 16 , wherein the NK cell is an NK-92 cell or NK-92MI cell.
18. A method of treating a subject with a coronavirus infection, comprising administering an effective amount of the modified immune cell of claim 14 to the subject.
19. The method of claim 18 , wherein the subject is infected with SARS-COV-1 or SARS-COV-2.