Non-viral chimeric antigen receptor (CAR) T cells going viral

H Balke-Want; V Keerthi; A Cadinanos-Garai; C Fowler; N Gkitsas; A K Brown; R Tunuguntla; M Abou-El-Enein; S A Feldman

doi:10.1016/j.iotech.2023.100375

Non-viral chimeric antigen receptor (CAR) T cells going viral

Immunooncol Technol. 2023 Mar 9:18:100375. doi: 10.1016/j.iotech.2023.100375. eCollection 2023 Jun.

Authors

H Balke-Want¹, V Keerthi¹, A Cadinanos-Garai², C Fowler¹, N Gkitsas¹, A K Brown¹, R Tunuguntla¹, M Abou-El-Enein^{2

3}, S A Feldman¹

Affiliations

¹ Stanford Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University, Stanford, USA.
² USC/CHLA Cell Therapy Program, University of Southern California, and Children's Hospital Los Angeles, Los Angeles, USA.
³ Division of Medical Oncology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, USA.

Abstract

Chimeric antigen receptor (CAR) T cell therapy has made significant strides in the treatment of B-cell malignancies, but its application in treating solid tumors still poses significant challenges. Particularly, the widespread use of viral vectors to deliver CAR transgenes into T cells comes with limitations, including high costs and regulatory restrictions, which hinder the translation of novel genetic engineering concepts into clinical applications. Non-viral methods, such as transposon/transposase and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas systems, offer promising alternatives for stable transgene insertion in CAR-T cells. These methods offer the potential to increase accessibility and efficiency in the development and delivery of CAR-T cell therapies. The main challenge in using non-viral methods, however, is their low knock-in efficiency, which leads to low transgene expression levels. In this review, we discuss recent developments in non-viral approaches for CAR-T cell production, the manufacturing requirements for clinical-grade production of non-viral CAR-T cells, and the adjustments needed in quality control for proper characterization of genomic features and evaluation of potential genotoxicity.

Keywords: CRISPR; cell therapy scaling; electroporation; gene delivery; manufacturing; non-viral CAR T cells.

Publication types

Review