Abstract Details

G2 - Immune Targeting and Approaches with Genetically-Modified Cells and Cell Therapies (Including CAR-T, CAR-NK, TCR editing)

830: Metabolic Priming of GD2 TRAC-CAR T Cells During Manufacturing Promotes Memory Phenotypes While Enhancing Persistence

Type: Poster Session

Poster Board Number: 830

Presentation Details

Session Title: Wednesday Posters: Immune Targeting and Approaches with Genetically-Modified Cells and Cell Therapies






Manufacturing Chimeric Antigen Receptor (CAR) T-cell therapies is complex, with a limited understanding of how media composition impacts T cell phenotypes. CRISPR/Cas9 ribonucleoproteins can precisely insert a CAR sequence while disrupting the endogenous T cell receptor alpha constant (TRAC) gene, resulting in TRAC-CAR T cells with an enriched stem cell memory T-cell population, a process that could be further optimized through modifications to the media composition. In this study we generated anti-GD2 TRAC-CAR T cells using ""metabolic priming"" (MP), where the cells were activated in glucose/glutamine low media and then expanded in glucose/glutamine high media. T cell products were evaluated using spectral flow cytometry, metabolic assays, cytokine production, cytotoxicity assays in vitro and potency against human GD2+ xenograft neuroblastoma models in vivo. Compared to standard TRAC-CAR T cells, MP TRAC-CAR T cells showed less glycolysis, higher CCR7/CD62L expression, more bound NAD(P)H activity and reduced IFN-γ, IL-2, IP-10, IL-1β, IL-17, and TGFβ production at the end of manufacturing ex vivo, with increased central memory CAR T cells and better persistence observed in vivo. Metabolic priming with media during CAR T cell biomanufacturing can minimize glycolysis and enrich memory phenotypes ex vivo, which could lead to better responses against solid tumors in vivo.

Plain Language Summary

CAR T cells were manufactured virus-free at scale with CRISPR/Cas9 and ‘metabolically primed’ by attenuating activation for three days in low glucose/glutamine media with further expansion in high glucose/glutamine media. This priming endowed CAR T-cells with properties of stem cell memory subsets, including enriched central memory phenotypes in vivo while lysing solid tumors.

Dan Cappabianca¹, Dan L. Pham¹, Matthew H. Forsberg¹, Anna Tommasi¹, Tony Lauer², Jolanta Vidugiriene², Brooke Hrdlicka¹, Alexandria McHale¹, Quaovi Sodji¹, Melissa C. Skala³, Christian M. Capitini¹, Krishanu Saha<sup>1

1</sup>University of Wisconsin-Madison, Madison, WI,²Promega Corporation, Fitchburg, WI,³Morgridge Institute for Research, Madison, WI"