Chimeric antigen receptor effector cell switches with humanized targeting moieties and/or optimized chimeric antigen receptor interacting domains and uses thereof
The present disclosure provides compositions, methods, kits, and platforms for selectively activating and deactivating chimeric receptor effector cells using humanized chimeric receptor effector cell switches that comprise a humanized targeting moiety that binds CD19 on a target cell and a chimeric receptor interacting domain that binds to a chimeric receptor effector cell and/or chimeric receptor effector cell switches comprising optimized chimeric receptor interacting domains. Also disclosed are methods of treating disease and conditions with such chimeric receptor effector cells and chimeric receptor effector cell switches.
1. A chimeric antigen receptor (CAR) comprising an extracellular domain, a transmembrane domain, and an intracellular signaling domain; wherein the extracellular domain comprises a humanized anti-GCN4 scFv comprising light chain CDRs 1-3 and heavy chain CDRs 1-3 respectively set forth as GSSTGAVTTSNYAS (SEQ ID NO:425), GTNNRAP (SEQ ID NO:426), VLWYSDHWV (SEQ ID NO:427), GFLLTDYGVN (SEQ ID NO:428), VIWGDGITD (SEQ ID NO:429), and GLFDY (SEQ ID NO:430); and wherein the extracellular domain optionally comprises a hinge domain.
2. The CAR of claim 1 , wherein the scFv comprises the amino acid sequence shown in SEQ ID NO:322.
3. The CAR of claim 1 , wherein the transmembrane domain comprises CD28TM, the intracellular signaling domain comprises CD28, 4-1BB, and CD3, and the hinge domain comprises an IgG4m hinge comprising SEQ ID NO:168.
4. The CAR of claim 1 , wherein the CAR comprises a sequence selected from the group consisting of SEQ ID NOS: 401, 403, 405, 407, 409, 411, 413, and 415.
5. The CAR of claim 1 , wherein the CAR comprises the amino acid sequence SEQ ID NO: 411.
6. The CAR of claim 1 , wherein the extracellular domain comprises a hinge domain comprising a sequence selected from the group consisting of SEQ ID NOS: 93-103 and 165-168.
7. The CAR of claim 1 , wherein the extracellular domain comprises a hinge domain comprising an amino acid sequence: ESKYGPPCPPCPD (SEQ ID NO:424).
8. The CAR of claim 1 , wherein the transmembrane domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 417 and 398.
9. The CAR of claim 1 , wherein the intracellular signaling domain comprises (a) a CD3-zeta domain, and (b) a CD28 domain; a 4-1BB domain; or a CD28 domain and a 4-1BB domain.
10. The CAR of claim 9 , wherein the CD28 domain comprises an amino acid sequence that is at least 95% identical to sequence SEQ ID NO: 418.
11. The CAR of claim 9 , wherein the CD28 domain comprises an amino acid sequence SEQ ID NO: 418.
12. The CAR of claim 9 , wherein the 4-1BB domain comprises an amino acid sequence that is at least 95% identical to sequence SEQ ID NO: 419.
13. The CAR of claim 9 , wherein the 4-1BB domain comprises an amino acid sequence SEQ ID NO: 419.
14. The CAR of claim 9 , wherein the CD3-zeta domain comprises an amino acid sequence that is at least 95% identical to sequence SEQ ID NO: 420.
15. The CAR of claim 9 , wherein the CD3-zeta domain comprises an amino acid sequence SEQ ID NO: 420.
16. The CAR of claim 1 , wherein the transmembrane domain comprises an amino acid sequence that is at least 95% identical to sequence SEQ ID NO: 417.
17. The CAR of claim 1 , wherein the transmembrane domain comprises an amino acid sequence SEQ ID NO: 417.