Sort-purification of human CD34+CD90+ cells reduces target cell population and improves lentiviral transduction

Radtke, S.; Pande, D.; Cui, M.; Perez, A. M.; Chan, Y.-Y.; Enstrom, M.; Schmuck, S.; Berger, A.; Eunson, T.; Adair, J. E.; Kiem, H.-P.

doi:10.1101/850479

bioRxiv · 10.1101/850479

Sort-purification of human CD34+CD90+ cells reduces target cell population and improves lentiviral transduction

Radtke, S.·Pande, D.·Cui, M.·Perez, A. M.·Chan, Y.-Y.·Enstrom, M.·Schmuck, S.·Berger, A.·Eunson, T.·Adair, J. E.·Kiem, H.-P.

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Abstract

ABSTRACTHematopoietic stem cell (HSC) gene therapy has the potential to cure many genetic, malignant and infectious diseases. We have shown in a nonhuman primate (NHP) HSC gene therapy and transplantation model that the CD34+CD90+ cell fraction was exclusively responsible for multilineage engraftment and hematopoietic reconstitution. Here we show the translational potential of this HSC-enriched CD34 subset for lentivirus-mediated gene therapy. Alternative HSC-enrichment strategies include the purification of CD133+ cells or CD38low/- subsets of CD34+ cells from human blood products. We directly compared these strategies to the isolation of CD90+ cells using a GMP-grade flow-sorting protocol with clinical applicability. We show that CD90+ cell selection results in 40-fold fewer target cells in comparison to CD133+ or CD38low/- CD34 subsets without compromising the engraftment potential in vivo. Single cell RNA sequencing confirmed nearly complete depletion of lineage committed progenitor cells in CD90+ fractions compared to alternative selections. Importantly, lentiviral transduction efficiency in purified CD90+ cells resulted in up to 3-fold higher levels of engrafted gene-modified blood cells. These studies should have important implications for the manufacturing of patient-specific HSC gene therapy and genome editing products.

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Radtke, S., Pande, D., Cui, M., Perez, A. M., Chan, Y.-Y., Enstrom, M., Schmuck, S., Berger, A., Eunson, T., Adair, J. E., Kiem, H.-P.. 2019-11-26. Sort-purification of human CD34+CD90+ cells reduces target cell population and improves lentiviral transduction. https://doi.org/10.1101/850479

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