Author:
Prochazkova Michaela,Dreyzin Alexandra,Shao Lipei,Garces Pam,Cai Yihua,Shi Rongye,Pelayo Alejandra,Kim Yong Soo,Pham Victoria,Frodigh Sue Ellen,Fenton Shannon,Karangwa Catherine,Su Yan,Martin Kathryn,Zhang Nan,Highfill Steven L.,Somerville Robert P.,Shah Nirali N.,Stroncek David F.,Jin Ping
Abstract
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.
Funder
National Institutes of Health
Publisher
Springer Science and Business Media LLC
Cited by
1 articles.
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