AI Article Synopsis
- * A new lentiviral transduction protocol was developed to successfully manipulate long-term repopulating quiescent HSC by addressing barriers like vector entry and limited resource availability.
- * This enhanced method, using cyclosporin H and deoxynucleosides, results in effective level of genetic modifications and improved HSC engraftment compared to traditional culture techniques, paving the way for future genetic engineering strategies.
Article Abstract
Quiescent human hematopoietic stem cells (HSC) are ideal targets for gene therapy applications due to their preserved stemness and repopulation capacities; however, they have not been exploited extensively because of their resistance to genetic manipulation. We report here the development of a lentiviral transduction protocol that overcomes this resistance in long-term repopulating quiescent HSC, allowing their efficient genetic manipulation. Mechanistically, lentiviral vector transduction of quiescent HSC was found to be restricted at the level of vector entry and by limited pyrimidine pools. These restrictions were overcome by the combined addition of cyclosporin H (CsH) and deoxynucleosides (dNs) during lentiviral vector transduction. Clinically relevant transduction levels were paired with higher polyclonal engraftment of long-term repopulating HSC as compared with standard ex vivo cultured controls. These findings identify the cell-intrinsic barriers that restrict the transduction of quiescent HSC and provide a means to overcome them, paving the way for the genetic engineering of unstimulated HSC.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10787167 | PMC |
| http://dx.doi.org/10.1016/j.ymthe.2023.11.020 | DOI Listing |
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