Journal of Translational Medicine (Apr 2024)

Deciphering the importance of culture pH on CD22 CAR T-cells characteristics

  • Michaela Prochazkova,
  • Alexandra Dreyzin,
  • Lipei Shao,
  • Pam Garces,
  • Yihua Cai,
  • Rongye Shi,
  • Alejandra Pelayo,
  • Yong Soo Kim,
  • Victoria Pham,
  • Sue Ellen Frodigh,
  • Shannon Fenton,
  • Catherine Karangwa,
  • Yan Su,
  • Kathryn Martin,
  • Nan Zhang,
  • Steven L. Highfill,
  • Robert P. Somerville,
  • Nirali N. Shah,
  • David F. Stroncek,
  • Ping Jin

DOI
https://doi.org/10.1186/s12967-024-05197-5
Journal volume & issue
Vol. 22, no. 1
pp. 1 – 15

Abstract

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Abstract Background Chimeric antigen receptor (CAR) T-cells have demonstrated significant efficacy in targeting hematological malignancies, and their use continues to expand. Despite substantial efforts spent on the optimization of protocols for CAR T-cell manufacturing, critical parameters of cell culture such as pH or oxygenation are rarely actively monitored during cGMP CAR T-cell generation. A comprehensive understanding of the role that these factors play in manufacturing may help in optimizing patient-specific CAR T-cell therapy with maximum benefits and minimal toxicity. Methods This retrospective study examined cell culture supernatants from the manufacture of CAR T-cells for 20 patients with B-cell malignancies enrolled in a phase 1/2 clinical trial of anti-CD22 CAR T-cells. MetaFLEX was used to measure supernatant pH, oxygenation, and metabolites, and a Bio-Plex assay was used to assess protein levels. Correlations were assessed between the pH of cell culture media throughout manufacturing and cell proliferation as well as clinical outcomes. Next-generation sequencing was conducted to examine gene expression profiles of the final CAR T-cell products. Results A pH level at the lower range of normal at the beginning of the manufacturing process significantly correlated with measures of T-cell expansion and metabolism. Stable or rising pH during the manufacturing process was associated with clinical response, whereas a drop in pH was associated with non-response. Conclusions pH has potential to serve as an informative factor in predicting CAR T-cell quality and clinical outcomes. Thus, its active monitoring during manufacturing may ensure a more effective CAR T-cell product.

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