Genetically-modified cells comprising a modified human T cell receptor alpha constant region gene
Disclosed herein is a genetically-modified cell comprising in its genome a modified human T cell receptor alpha constant region gene, wherein the cell has reduced cell-surface expression of the endogenous T cell receptor. The present disclosure further relates to methods for producing such a genetically-modified cell, and to methods of using such a cell for treating a disease in a subject.
1. A genetically-modified human T cell comprising in its genome a modified human T cell receptor (TCR) alpha constant region gene, wherein said modified human TCR alpha constant region gene comprises from 5′ to 3′:
(a) a 5′ region of a human TCR alpha constant region gene which is endogenous to said T cell;
(b) an exogenous nucleic acid sequence encoding a chimeric antigen receptor; and
(c) a 3′ region of said human TCR alpha constant region gene which is endogenous to said T cell;
wherein said exogenous nucleic acid sequence is inserted in said TCR alpha constant region gene which is endogenous to said T cell at a position within SEQ ID NO: 3, and wherein said chimeric antigen receptor comprises a ligand-binding domain having specificity for an antigen present on a cancer cell, and wherein said genetically-modified human T cell expresses said chimeric antigen receptor and exhibits reduced cell-surface expression of the endogenous T cell receptor when compared to unmodified control cells.
2. The cell of claim 1 , wherein said exogenous nucleic acid sequence comprises a promoter sequence that drives expression of said chimeric antigen receptor.
3. The cell of claim 1 , wherein said chimeric antigen receptor comprises an intracellular cytoplasmic signaling domain having at least 95% sequence identity to SEQ ID NO: 113.
4. The cell of claim 1 , wherein the ligand-binding domain of said chimeric antigen receptor is specific for CD19.