Chimeric antigen receptors, compositions, and methods
This disclosure describes chimeric antigen receptors for expression in a Natural Killer (NK) cell, pharmaceutical compositions that include NK cells (and/or iPSCs) modified to express a chimeric antigen receptor, and methods involving such chimeric antigen receptors. Generally, the chimeric antigen receptor includes an ectodomain that includes an antigen recognition region, a transmembrane domain linked to the ectodomain, and an endodomain linked to the transmembrane domain. The endodomain can include a signaling peptide that activates an NK cell.
1. A chimeric antigen receptor molecule comprising, in an N-terminus to C-terminus orientation: an ectodomain comprising an antigen recognition region; a hinge; a transmembrane domain comprising a transmembrane region of NKG2D (cluster of differentiation 314 or CD314) in reverse orientation as compared to native NKG2D that natively has an extracellular C-terminus; and an endodomain comprising a functional signaling domain that activates a Natural Killer (NK) cell, wherein said functional signaling domain is selected from the group consisting of:
(a) a 2B4 (cluster of differentiation 244 or CD244) intracellular domain (ICD) and a CD3 zeta (CD3ζ) signaling domain;
(b) a 41BB (cluster of differentiation 137 or CD137) ICD and a CD3ζ signaling domain;
(c) a DAP12 (DNAX activation protein of 12 kDa) ICD and a CD3ζ signaling domain;
(d) a 2B4 ICD, a 41BB ICD and a CD3ζ signaling domain;
(e) a IL21R ICD and a CD3ζ signaling domain; and
(f) a 41BB ICD, a 2B4 ICD and a CD3ζ signaling domain.
2. The chimeric antigen receptor of claim 1 wherein the antigen recognition domain specifically binds an antigen associated with a disease.
3. The chimeric antigen receptor of claim 1 wherein the antigen recognition domain specifically binds a tumor antigen.
4. The chimeric antigen receptor of claim 1 wherein the ectodomain further comprises a signal peptide or leader sequence.
5. The chimeric antigen receptor of claim 1 , wherein said hinge is a CD8α hinge.
6. A pharmaceutical composition comprising an NK cell that expresses the chimeric antigen receptor of claim 5 .
7. The pharmaceutical composition of claim 6 , wherein the NK cell is derived from an iPSC.