US 12,187,776 B2
Pooling signaling and costimulatory domains in B7H6 chimeric antigen receptor
David Gilham, Mont-Saint-Guibert (BE); Jennifer Bolsée, Mont-Saint-Guibert (BE); and Lorraine Springuel, Mont-Saint-Guibert (BE)
Assigned to CELYAD S.A., Mont-Saint-Guibert (BE)
Appl. No. 16/772,183
Filed by CELYAD S.A, Mont-Saint-Guibert (BE)
PCT Filed Dec. 14, 2018, PCT No. PCT/EP2018/085089
§ 371(c)(1), (2) Date Jun. 12, 2020,
PCT Pub. No. WO2019/115818, PCT Pub. Date Jun. 20, 2019.
Claims priority of application No. 17207309 (EP), filed on Dec. 14, 2017.
Prior Publication US 2021/0070831 A1, Mar. 11, 2021
Int. Cl. C07K 14/725 (2006.01); A61K 39/00 (2006.01); A61P 35/00 (2006.01); C07K 14/705 (2006.01); C07K 16/28 (2006.01); C12N 5/0783 (2010.01)
CPC C07K 14/7051 (2013.01) [A61K 39/4611 (2023.05); A61K 39/4631 (2023.05); A61K 39/464429 (2023.05); A61P 35/00 (2018.01); C07K 14/705 (2013.01); C07K 16/2827 (2013.01); C12N 5/0636 (2013.01); C07K 2317/622 (2013.01); C07K 2319/02 (2013.01); C07K 2319/03 (2013.01)] 12 Claims
 
1. A combination of nucleic acid molecules which when co-expressed result in a functional chimeric receptor, which comprises:
(a) a first nucleic acid molecule which comprises a nucleic acid encoding an antigen binding domain, a hinge region, a nucleic acid encoding the NKG2D transmembrane (TM) domain or a nucleic acid encoding a polylysine of the same length as NKG2D TM which has been modified to comprise an arginine at position 11 or 12, wherein sequence numbering corresponds to that of the native NKG2D transmembrane domain, a nucleic acid encoding the cytoplasmic (CYP) domain of NKG2D, and a CD3zeta signaling domain and (b) a second nucleic acid molecule which encodes at least the TM region of DAP10 and further comprises the endogenous DAP10 signaling domain or a CD28 costimulatory domain;
wherein the TM domain encoded by the first nucleic acid associates with the TM domain of DAP10 encoded by the second nucleic acid; thereby resulting in a functional CAR, and wherein the antigen binding domain encoded by the first nucleic acid binds to B7H6.
 
3. A combination of nucleic acid molecules which comprises:
(a) a first nucleic acid molecule which comprises a nucleic acid encoding an antigen binding domain, a CD28 or CD8a hinge, a nucleic acid encoding the NKp44™, a nucleic acid encoding the cytoplasmic (CYP) domain of NK44, and a CD3zeta signaling domain; and
(b) a second nucleic acid molecule which encodes at least the TM region of DAP12 and further comprises the endogenous DAP12 signaling domain or a CD28 costimulatory domain;
wherein the TM domain encoded by the first nucleic acid of (a) associates with the TM domain of the DAP12 accessory protein encoded by the second nucleic acid of (b), thereby resulting in a functional CAR, and wherein the antigen binding domain encoded by the first nucleic acid binds to B7H6.