Assessing a single-cell multi-omic analytic platform to characterize ex vivo -engineered T-cell therapy products

• Published in: Frontiers in bioengineering and biotechnology 2024-08, Vol.12, p.1417070
• Main Authors: Moshref, Maryam, Lo, Jerry Hung-Hao, McKay, Andrew, Camperi, Julien, Schroer, Joseph, Ueno, Norikiyo, Wang, Shu, Gulati, Saurabh, Tarighat, Somayeh, Durinck, Steffen, Lee, Ho Young, Chen, Dayue
• Format: Article
• Language: English
• Subjects: Bioengineering and Biotechnology; clustered regularly interspaced short palindromic repeats; engineered CD8+ T cells; final cell product; heterogeneity; multi-omics; single-cell
• ISSN: 2296-4185; 2296-4185
• DOI: 10.3389/fbioe.2024.1417070

Genetically engineered CD8 T cells are being explored for the treatment of various cancers. Analytical characterization represents a major challenge in the development of genetically engineered cell therapies, especially assessing the potential off-target editing and product heterogeneity. As conventional sequencing techniques only provide information at the bulk level, they are unable to detect off-target CRISPR translocation or editing events occurring in minor cell subpopulations. In this study, we report the analytical development of a single-cell multi-omics DNA and protein assay to characterize genetically engineered cell products for safety and genotoxicity assessment. We were able to quantify on-target edits, off-target events, and potential translocations at the targeting loci with per-cell granularity, providing important characterization data of the final cell product. Conclusion: A single-cell multi-omics approach provides the resolution required to understand the composition of cellular products and identify critical quality attributes (CQAs).