NKP30 receptor targeted therapeutics
The invention is directed to T cells and other cells that express chimeric NK-p30 receptors (“chimeric NKp30 T cells”), methods of making and using chimeric NKp30 T cells, and methods of using these chimeric NKp30 T cells, isolated populations thereof, and compositions comprising the same. In another aspect, said chimeric NKp30 T cells are further designed to express a functional non-TCR receptor. The disclosure also pertains to methods of making said chimeric NKp30 T cells, and methods of reducing or ameliorating, or preventing or treating, diseases and disorders using said chimeric NKp30 T cells, populations thereof, or compositions comprising the same.
1. A nucleic acid construct that encodes a chimeric receptor, or a cell containing said nucleic acid construct, wherein said nucleic acid construct comprises a promoter operably linked to a nucleic acid sequence that encodes a chimeric NKp30 receptor comprising an NKp30 extracellular domain, a transmembrane domain, and at least one signaling domain, wherein (i) said transmembrane domain comprises a transmembrane domain of a polypeptide selected from the group consisting of NKp30, CD28, CD8, CD3ζ, CD27, 41BB, OX40, and DAP12, and (ii) said at least one signaling domain comprises a signaling domain of a polypeptide selected from the group consisting of NKp30, CD28, CD8, CD3ζ, DAP10, CD27, 41BB, OX40, and DAP12, wherein at least one transmembrane (TM) domain or at least one signaling domain is from a polypeptide other than NKp30.
2. A chimeric polypeptide encoded by the nucleic acid construct of claim 1 .
3. The chimeric polypeptide of claim 2 , comprising
(i) a NKp30 extracellular domain, a CD28 transmembrane (TM) domain, a CD28 signaling domain, and a CD3ζ signaling domain;
(ii) a chimeric NKp30-CD28-CD3ζ receptor comprising the amino acid sequence of SEQ ID NO:61;
(iii) a NKp30 extracellular domain, a CD3ζ transmembrane domain, and a CD3ζ signaling domain;
(iv) a chimeric NKp30-CD3ζ receptor comprising the amino acid sequence of SEQ ID NO:60;
(v) a NKp30 extracellular domain, a CD28 transmembrane (TM) domain, a DAP10 signaling domain, and a CD3ζ signaling domain;
(vi) a chimeric NKp30-CD28(TM)-DAP10-CD3ζ receptor comprising the amino acid sequence of SEQ ID NO: 65;
(vii) a NKp30 extracellular domain, a CD28 transmembrane (TM) domain, a DAP10 signaling domain, and a CD3ζ signaling domain;
(viii) a chimeric NKp30-CD28(TM)-CD3ζ-DAP10 receptor comprising the amino acid sequence of SEQ ID NO: 67;
(ix) a NKp30 extracellular domain, a CD28 transmembrane (TM) domain, a CD27 signaling domain, and a CD3ζ signaling domain;
(x) a chimeric NKp30-CD28(TM)-CD27-CD3ζ receptor comprising the amino acid sequence of SEQ ID NO: 69;
(xi) a NKp30 extracellular domain, a CD28 transmembrane (TM) domain, and a CD3ζ signaling domain;
(xii) a chimeric NKp30-CD28(TM)-CD3ζ receptor comprising the amino acid sequence of SEQ ID NO: 62;
(xiii) a NKp30 extracellular domain, a CD8 transmembrane (TM) domain, and a CD3ζ signaling domain; or
(xiv) a chimeric NKp30-CD8(TM)-CD3ζ receptor comprising the amino acid sequence of SEQ ID NO: 63.
4. The nucleic acid construct according to claim 1 , which encodes
(i) a chimeric NKp30-CD3ζ receptor, said nucleic acid construct comprising the nucleic acid sequence of SEQ ID NO:124;
(iii) a chimeric NKp30-CD28(TM)-CD3ζ-DAP10 receptor, said nucleic acid construct comprising the nucleic acid sequence of SEQ ID NO:66;
(iv) a chimeric NKp30-CD28(TM)-CD27-CD3ζ receptor, said nucleic acid construct comprising the nucleic acid sequence of SEQ ID NO: 68;
(v) a chimeric NKp30-CD8(TM)-CD3ζ receptor, said nucleic acid construct comprising the nucleic acid sequence or SEQ ID NO: 127;
(vi) a chimeric NKp30-CD28-CD3ζ receptor, said nucleic acid construct comprising the nucleic acid sequence of SEQ ID NO:125; or
(vii) a chimeric NKp30-CD28(TM)-CD3ζ receptor, said nucleic acid construct comprising the nucleic acid sequence of SEQ ID NO: 126.
5. A chimeric receptor polypeptide expressed by the nucleic acid construct of claim 4 .
6. A T cell which comprises a nucleic acid construct according to claim 1 .
7. A T cell which comprises a nucleic acid construct according to claim 4 .
8. The T cell of claim 6 which comprises a primary human T cell.
9. The T cell of claim 7 which comprises a primary human T cell.
10. A composition containing a T cell according to claim 6 .
11. A composition containing a T cell according to claim 7 .
12. The chimeric polypeptide of claim 2 , which further comprises an additional signaling domain comprising a signaling domain of a polypeptide selected from the group consisting of NKp30, CD28, CD8, CD3ζ, DAP10, CD27, 41BB, OX40, and DAP12.
13. The chimeric polypeptide of claim 2 , which further comprises an additional CD28 signaling domain.
14. The chimeric polypeptide of claim 2 , comprising at least one of the following:
(i) a NKp30 extracellular domain as shown in FIG. 16 , FIG. 17 , FIG. 18 , FIG. 20 , FIG. 21 , or FIG. 22 ;
(ii) a CD3ζ transmembrane (TM) domain as shown in FIG. 16 ;
(iii) a CD28 transmembrane (TM) domain as shown in FIG. 18 , FIG. 20 , FIG. 21 , or FIG. 22 ;
(iv) a CD3ζ signaling domain that comprises the CD3ζ cytoplasmic domain as shown in FIG. 18 , FIG. 20 , FIG. 21 , or FIG. 22 ;
(v) a DAP10 signaling domain that comprises the DAP10 cytoplasmic domain as shown in FIG. 20 or FIG. 21 ; or
(vi) a CD27 signaling domain that comprises the CD27 cytoplasmic domain shown in FIG. 22 .
15. The nucleic acid construct, or cell containing said nucleic acid construct, according to claim 1 , wherein the nucleic acid construct further comprises a suicide gene.