Altering gene expression in modified T cells and uses thereof
AssigneeThe Trustees of the University of Pennsylvania
InventorsYangbing Zhao, Jiangtao Ren, Xiaojun Liu, Carl H. June
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Abstract
The present invention relates to compositions and methods for generating a modified T cell with a nucleic acid capable of downregulating endogenous gene expression selected from the group consisting of TCR α chain, TCR β chain, beta-2 microglobulin and FAS further comprising a nucleic acid encoding a modified T cell receptor (TCR) comprising affinity for a surface antigen on a target cell or an electroporated nucleic acid encoding a chimeric antigen receptor (CAR). Also included are methods and pharmaceutical compositions comprising the modified T cell for adoptive therapy and treating a condition, such as an autoimmune disease.
Claims19
A method of treating a condition associated with enhanced immunity in a subject, comprising administering to the subject a therapeutically 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 gene locus causing a downregulation of the endogenous gene expression, wherein the endogenous gene is selected from the group consisting of TCR α chain (TRAC), TCR β chain (TRBC), beta-2 microglobulin (B2M), a HLA molecule, CTLA-4, PD1, and FAS;
- wherein the CRISPR-mediated insertion or deletion is within:
- (i) the TCR α chain gene locus and the TCR β chain gene locus, causing the downregulation of the TCR α chain gene expression and the TCR β chain gene expression, and wherein the CRISPR-mediated insertion or deletion in the TCR α chain gene locus is within a TCR α chain nucleotide sequence comprising SEQ ID NO:
- 1;
- and
- (ii) the beta-2 microglobulin (B2M) gene locus, causing the downregulation of the B2M gene expression;
- and wherein the CRISPR-mediated insertion or deletion is within a B2M gene nucleotide sequence comprising SEQ ID NO:
- 3, 44, 77, or 78;
- and
- (b) a nucleic acid encoding an exogenous T cell receptor (TCR) having an affinity for a surface antigen on a target cell, wherein the exogenous TCR comprises a TCR alpha chain and TCR beta chain.
A method of treating a condition associated with enhanced immunity in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising:
- (a) a CRISPR-modified T cell, wherein the CRISPR-modified T cell comprises:
- (i) a CRISPR-mediated insertion or deletion in an endogenous TCR α chain (TRAC) gene locus and an endogenous TCR β chain (TRBC) gene locus, causing downregulation of the endogenous TCR α chain (TRAC) and TCR β chain (TRBC) genes expression, wherein the CRISPR-mediated insertion or deletion in the TRAC gene locus is are within a TRAC nucleotide sequence comprising SEQ ID NO:
- 1;
- and
- (ii) a CRISPR-mediated insertion or deletion in an endogenous beta-2 microglobulin (B2M) gene locus, causing downregulation of the B2M gene expression;
- and wherein the CRISPR-mediated insertion or deletion is within a B2M nucleotide sequence comprising SEQ ID NO:
- 3, 44, 77, or 78;
- and
- (b) a nucleic acid encoding a modified T cell receptor (TCR) having an affinity for NY-ESO-1 on a target cell.
Dependencies are inferred from the claim text.