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A single cell RNA analytical platform for characterizing cell states in gene therapy drug products

August 11 @ 9:00 am - 10:00 am PDT

Human hematopoietic/stem progenitor (HSPC) CD34+ cells are the core components of ex vivo gene therapy (GT) for the treatment of rare monogenic diseases. Assessment of CD34+ cells for their long-term hematopoietic stem cell (HSC) content before and after ex vivo transduction, as well as in vivo after transplantation, is a critical parameter to establish the efficacy of GT. To overcome the limitations of standard immunophenotyping, we leveraged the high resolution offered by single cell (sc)-RNA-sequencing to capture the heterogeneity of HSPC states and its dynamic changes. Through integrated analysis of the scRNA of individually cultured HSPC subsets and bulk CD34+ before and after in vitro manipulation (total of 176,250 single cells), we unveiled the impact of cell culture conditions on the composition of the GT drug product. We anticipate such information will allow us to generate a robust protocol for the manufacture of transduced CD34+ cells for HSPC GT.