Granted Patent B2

US 12,570,982 · App. 17/550,455 · Granted Mar 10, 2026

Inventors: Yangbing Zhao (Lumberton, NJ); Jiangtao Ren (Philadelphia, PA); Xiaojun Liu (Wallingford, PA); Carl H. June (Merion Station, PA)

Assignee: THE TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA

C12N15/1138A61K35/17A61K35/26A61K39/001102A61K40/11A61K40/22A61K40/31A61K40/32A61K40/36A61K40/416A61K40/418A61K40/4211A61K40/4269A61K40/4274A61P35/00A61P37/06C12N5/0636C12N15/85A61K2039/5156A61K2039/5158A61K40/50C12N2310/10C12N2310/20C12N2501/48C12N2501/515C12N2501/599C12N2501/998C12N2510/00

Abstract

Claims (27)

1 . A method for stimulating a T cell-mediated immune response to a target cell or tissue in a subject comprising:

(i) administering to the subject an effective amount of a pharmaceutical composition comprising a CRISPR-modified T cell, wherein the CRISPR-modified T cell comprises:

(a) a CRISPR-mediated insertion or deletion in an endogenous TCR a chain (TRAC) gene locus capable of downregulating gene expression of the endogenous TCR a chain gene, wherein the CRISPR-mediated insertion or deletion is within a TCR a chain nucleotide sequence comprising SEQ ID NO: 1;

(b) a CRISPR-mediated insertion or deletion in an endogenous TCR β chain (TRBC) gene locus capable of downregulating gene expression of the endogenous TCR B chain gene;

(c) a CRISPR-mediated insertion or deletion in an endogenous beta-2-microglobulin (B2M) gene locus capable of downregulating gene expression of the endogenous B2M gene, wherein the CRISPR-mediated insertion or deletion is within a B2M nucleotide sequence comprising SEQ ID NO: 3, 44, 77, or 78; and

(d) a nucleic acid encoding an exogenous T cell receptor (TCR) comprising affinity for a tumor associated antigen (TAA) on the target cell or tissue;

and

(ii) inducing lysis of the target cell or tissue by administering to the subject an effective amount of a pharmaceutical composition comprising an antibody that selectively blocks CTLA-4, PD-1 and/or PD-L1.

2 . The method of claim 1 , wherein the induced lysis is antibody-dependent cell-mediated cytotoxicity (ADCC).

3 . The method of claim 1 , wherein the CRISPR-mediated insertion or deletion comprises use of a CRISPR system comprising a pAd5/F35-CRISPR vector.

4 . The method of claim 1 , wherein the exogenous TCR is selected from the group consisting of a wild-type TCR, a modified TCR, and a chimeric TCR.

5 . The method of claim 4 , wherein the modified TCR comprises at least one disulfide bond.

6 . The method of claim 4 , wherein the modified TCR comprises a TCR alpha chain and a TCR beta chain.

7 . The method of claim 6 , wherein the modified TCR comprises a co-stimulatory signaling domain at a C terminus of at least one of the TCR alpha chain and the TCR beta chain.

8 . The method of claim 7 , wherein the co-stimulatory signaling domain is a 4-1BB co-stimulatory signaling domain.

9 . The method of claim 6 , wherein the TCR beta chain comprises at least one N-deglycosylation.

10 . The method of claim 6 , wherein the TCR alpha chain comprises at least one N-deglycosylation.

11 . The method of claim 4 , wherein the modified TCR comprises at least one murine constant region.

12 . The method of claim 4 , wherein the modified TCR has higher affinity for the TAA when compared to the wild-type TCR.

13 . The method of claim 1 , wherein the CRISPR-modified T cell further comprises a CRISPR-mediated insertion or deletion in an endogenous gene locus selected from the group consisting of a HLA molecule, CTLA-4, PD1, and FAS, and wherein the CRISPR-mediated insertion or deletion downregulates the gene expression of the HLA molecule, CTLA-4, PD1, and FAS.

14 . The method of claim 1 , wherein the CRISPR-modified T cell further comprises a CRISPR-mediated insertion or deletion in an endogenous PD1 gene locus capable of downregulating gene expression of the endogenous PD1 gene.

15 . A method for stimulating a T cell-mediated immune response to a target cell or tissue in a subject comprising administering to the subject an effective amount of a pharmaceutical composition comprising a CRISPR-modified T cell, and a pharmaceutical composition comprising an antibody that selectively blocks CTLA-4, PD-1 and/or PD-L1;

wherein the CRISPR-modified T cell comprises:

(a) a CRISPR-mediated insertion or deletion in an endogenous TCR α chain (TRAC) gene locus capable of downregulating gene expression of the endogenous TCR α chain gene, wherein the CRISPR-mediated insertion or deletion is in a TCR α chain nucleotide sequence comprising SEQ ID NO: 1;

(b) a CRISPR-mediated insertion or deletion in an endogenous TCR β chain (TRBC) gene locus capable of downregulating gene expression of the endogenous TCR β chain gene;

(c) a CRISPR-mediated insertion or deletion in an endogenous B2M gene locus capable of downregulating gene expression of the B2M gene, wherein the CRISPR-mediated insertion or deletion is within a B2M nucleotide sequence comprising SEQ ID NO: 3, 44, 77, or 78; and

(d) a nucleic acid encoding a modified NY-ESO-1 T cell receptor (TCR), wherein the modified NY-ESO-1 TCR is an 8F NY-ESO-1 TCR.

Assignments (1)

Continuity (3)

References Cited (242)