Development of a non-viral gene delivery platform for CAR-T manufacturing

The use of CAR-T cells as potent therapies for refractory B-cell and plasma cell malignancies poses an immense financial burden. It requires lengthy manufacturing times, especially for GMP-grade vector production.

Non-viral gene delivery methods are therefore a desirable alternative, and the first non-viral cell therapy was recently FDA-approved. Homology-directed recombination (HDR) is a common strategy for gene delivery, but its low frequency yields few CAR-T cells during manufacturing. In contrast, non-homologous end joining (NHEJ) is the primary pathway of dsDNA break repair, and NHEJ-mediated homology-independent targeted insertion (HITI) has been previously shown to more efficiently knock-in targets as compared to HDR in several cell types and pre-clinical studies.

Here, we show HITI-mediated site-directed integration of a therapeutically relevant GD2-CAR transgene into the T cell receptor alpha constant (TRAC) locus using nanoplasmid DNA and CRISPR/Cas9 in primary human T cells. We will compare viral and non-viral platforms for developing a GD2 CAR-T manufacturing process while demonstrating process comparability and non-viral process feasibility. We will provide a deeper understanding of how to improve cell engineering efficiencies, cell viabilities, and scalability. 

• Explore the cutting-edge techniques employed by leading institutions increasing CAR knock-ins in T cells mediated by by homology-directed repair (HDR) or homology-independent targeted insertion (HITI)
• Explore how selecting a clinically-validated manufacturing platform for crucial processes like cell electroporation can mitigate risks in cell therapy production
• Gain a deeper understanding of how to use a non-viral process for CAR-T manufacturing

Steven Feldman

Site Head & Scientific Director of the Laboratory for Cell & Gene Medicine at Stanford Center for Cancer Cell Therapy

Dr Feldman is part of the Stanford Center for Cancer Cell Therapy and serves as the Site Head and Scientific Director of Stanford's GMP Facility. Here, he is leading a team that focuses on the development and manufacture of novel cell therapies for the treatment of cancer.

James Brady

Senior Vice President, Technical Applications & Customer Support at MaxCyte

Dr Brady is an experienced biotechnology industry professional with expertise in cell and gene therapy, biologics, and drug discovery. Previously, he was a Senior Scientist at Genetic Therapy, a Novartis subsidiary, where he worked on lentiviral-based gene therapy treatments, and was a Group Leader at MetaMorphix, managing the company’s transgenic and genetic research programs.

Dr Brady earned a Master of Business Administration degree in finance from The Johns Hopkins University. He completed his postdoctoral fellowship at the National Eye Institute of the National Institutes of Health in Bethesda, Maryland after obtaining a PhD in genetics from Indiana University in Bloomington.

Daniel Nguyen

Senior Manager, Global Sales Development & Inside Sales at MaxCyte

Daniel has extensive experience supporting industry and academic research labs with their electroporation and cell engineering-related needs from basic research to manufacturing. He has spent the last 8 years in the biotechnology field at companies like 23andMe, Synthego & MaxCyte that are focused on supporting scientists in the areas of cell therapy, CRISPR engineering, protein production, and disease modelling.

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SPEAKERS

Steven Feldman

Site Head & Scientific Director of the Laboratory for Cell & Gene Medicine at Stanford Center for Cancer Cell Therapy

James Brady

Senior Vice President, Technical Applications & Customer Support at MaxCyte

Daniel Nguyen

Senior Manager, Global Sales Development & Inside Sales at MaxCyte

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