Chimeric NK receptor and methods for treating cancer
The present invention relates to chimeric immune receptor molecules for reducing or eliminating tumors. The chimeric receptors are composed a C-type lectin-like natural killer cell receptor, or a protein associated therewith, fused to an immune signaling receptor containing an immunoreceptor tyrosine-based activation motif. Methods for using the chimeric receptors are further provided.
1. A nucleic acid construct comprising: a first nucleic acid sequence encoding a promoter operably linked to a second nucleic acid sequence encoding a chimeric receptor polypeptide, said second nucleic acid sequence comprising a nucleic acid encoding a C-type lectin-like type II natural killer cell receptor polypeptide fused to a nucleic acid encoding an immune signaling receptor polypeptide comprising SEQ ID NO:1, wherein when the nucleic acid construct is introduced into a T-lymphocyte, said chimeric receptor polypeptide is expressed on the surface of the T-lymphocyte, and said C-type lectin type II natural killer receptor polypeptide binds a ligand and activates said immune signaling receptor polypeptide comprising SEQ ID NO: 1.
2. The nucleic acid construct of claim 1 , wherein the construct is in a vector.
3. The nucleic acid construct of claim 1 , further comprising a suicide gene.
4. An isolated T cell comprising the nucleic acid construct of claim 1 .
5. An isolated T cell comprising the vector of claim 2 .
6. The nucleic acid construct of claim 1 , wherein the C-type lectin-like NK cell type II receptor is at the C-terminus of the chimeric receptor polypeptide and the immune signaling receptor polypeptide is at the N-terminus of the chimeric receptor polypeptide.
7. The nucleic acid construct of claim 1 , wherein the C-type lectin-like NK cell type II receptor is selected from Dectin-1, Mast cell function-associated antigen, HNKR-P1A, LLT1, CD69, CD69 homolog, CD72, CD94, KLRF1, Oxidized LDL receptor, CLEC-1, CLEC-2, NKG2D, NKG2C, NKG2A, NKG2E, and Myeloid DAP12-associating lectin.
8. The nucleic acid construct of claim 1 , wherein the C-type lectin-like NK cell type II receptor is human NKG2D having the sequence set forth in SEQ ID NO:2 or is human NKG2C having the sequence set forth in SEQ ID NO:3.
9. The nucleic acid construct of claim 1 , wherein the immune signaling receptor is CD3-zeta having the sequence set forth in SEQ ID NO:6, or is human Fc epsilon receptor-gamma chain having the sequence set forth in SEQ ID NO:7 or the cytoplasmic domain or a splicing variant thereof.
10. The nucleic acid construct of claim 1 , wherein the chimeric receptor polypeptide is a fusion between NKG2D and CD3-zeta.
11. The isolated T cell of claim 4 , which comprises a primary human T cell.
12. The isolated T cell of claim 5 , which comprises a primary human T cell.
13. The nucleic acid construct of claim 1 , wherein the C-type lectin-like NK cell type II receptor is human NKG2D.
14. The nucleic acid construct of claim 13 , which further comprises a suicide gene.
15. An isolated vector comprising the nucleic acid construct of claim 13 , which further comprises a nucleic acid encoding DAP10 or DAP12.
16. An isolated primary human T cell comprising the nucleic acid construct of claim 10 .
17. An isolated primary human T cell comprising the nucleic acid construct of claim 13 .
18. An isolated primary human T cell comprising the nucleic acid construct of claim 14 .
19. An isolated primary human T cell comprising the vector of claim 15 .