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Mol Ther. 2017 Apr 5;25(4):949-961. doi: 10.1016/j.ymthe.2017.02.005. Epub 2017 Feb 23.

Integration of a CD19 CAR into the TCR Alpha Chain Locus Streamlines Production of Allogeneic Gene-Edited CAR T Cells.

Author information

1
Precision BioSciences, Durham, NC 27701, USA. Electronic address: dan.macleod@precisionbiosciences.com.
2
Precision BioSciences, Durham, NC 27701, USA.
3
Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Ophthalmology, University of North Carolina, Chapel Hill, NC 27599; USA.
4
Precision BioSciences, Durham, NC 27701, USA. Electronic address: bruce.mccreedy@precisionbiosciences.com.

Abstract

Adoptive cellular therapy using chimeric antigen receptor (CAR) T cell therapies have produced significant objective responses in patients with CD19+ hematological malignancies, including durable complete responses. Although the majority of clinical trials to date have used autologous patient cells as the starting material to generate CAR T cells, this strategy poses significant manufacturing challenges and, for some patients, may not be feasible because of their advanced disease state or difficulty with manufacturing suitable numbers of CAR T cells. Alternatively, T cells from a healthy donor can be used to produce an allogeneic CAR T therapy, provided the cells are rendered incapable of eliciting graft versus host disease (GvHD). One approach to the production of these cells is gene editing to eliminate expression of the endogenous T cell receptor (TCR). Here we report a streamlined strategy for generating allogeneic CAR T cells by targeting the insertion of a CAR transgene directly into the native TCR locus using an engineered homing endonuclease and an AAV donor template. We demonstrate that anti-CD19 CAR T cells produced in this manner do not express the endogenous TCR, exhibit potent effector functions in vitro, and mediate clearance of CD19+ tumors in an in vivo mouse model.

KEYWORDS:

chimeric antigen receptor; gene editing; homology-directed repair

PMID:
28237835
PMCID:
PMC5383629
DOI:
10.1016/j.ymthe.2017.02.005
[Indexed for MEDLINE]
Free PMC Article

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