Journal Article

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

Hux, Jo Ann, Beard, Clayton W., Hirsch, Matthew L., Turner, Caitlin A., Lape, Janel, Wetzel, Keith J., Nicholson, Michael G., Hekele, Armin, Pham, Christina D., Brown, Audrey E., McCreedy, Bruce, Martin, Aaron J., Antony, Jeyaraj, Moser, Rachel J., Triggiano, Melissa A., Smith, Jeff, Jantz, Derek, Bartsevich, Victor V., MacLeod, Daniel T., Kirkland, Samantha

2017

Overview

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. MacLeod and colleagues use a homing endonuclease and an AAV vector to insert an anti-CD19 CAR into the TRAC gene to create universal allogeneic CAR T cells. Targeted insertion of a CAR transgene may allow the generation of more homogeneous CAR T cell products with more predictable safety and efficacy profiles.

• Publisher: Elsevier Inc,Elsevier Limited,American Society of Gene & Cell Therapy
• Subject: Alleles / Animals / Antigens / Antigens, CD19 - genetics / Autografts / Batch Cell Culture Techniques / CD19 antigen / Cell Engineering / chimeric antigen receptor / Chimeric antigen receptors / Clinical trials / CRISPR / Deoxyribonucleic acid / Dependovirus - genetics / Disease Models, Animal / DNA / Endonuclease / Experiments / Gene Editing / Gene Expression / Gene Knockout Techniques / Gene Order / Genetic Loci / Genetic Vectors - genetics / Genome editing / Genomes / Graft-versus-host reaction / Hematology / Homing endonuclease / homology-directed repair / Humans / Immunotherapy, Adoptive / Lymphocytes / Lymphocytes T / Lymphoma - genetics / Lymphoma - immunology / Lymphoma - therapy / Mice / Neoplasms / Original / Receptors, Antigen, T-Cell - genetics / Receptors, Antigen, T-Cell, alpha-beta - genetics / T cell receptors / T-Lymphocytes - immunology / T-Lymphocytes - metabolism / Transduction, Genetic / Tumors