Staurosporine Increases Lentiviral Vector Transduction Efficiency of Human Hematopoietic Stem and Progenitor Cells.

ABSTRACT: Lentiviral vector (LVV)-mediated transduction of human CD34⁺ hematopoietic stem and progenitor cells (HSPCs) holds tremendous promise for the treatment of monogenic hematological diseases. This approach requires the generation of a sufficient proportion of gene-modified cells. We identified staurosporine, a serine/threonine kinase inhibitor, as a small molecule that could be added to the transduction process to increase the proportion of genetically modified HSPCs by overcoming a LVV entry barrier. Staurosporine increased vector copy number (VCN) approximately 2-fold when added to mobilized peripheral blood (mPB) CD34⁺ cells prior to transduction. Limited staurosporine treatment did not affect viability of cells post-transduction, and there was no difference in in vitro colony formation compared to vehicle-treated cells. Xenotransplantation studies identified a statistically significant increase in VCN in engrafted human cells in mouse bone marrow at 4 months post-transplantation compared to vehicle-treated cells. Prostaglandin E₂ (PGE₂) is known to increase transduction efficiency of HSPCs through a different mechanism. Combining staurosporine and PGE₂ resulted in further enhancement of transduction efficiency, particularly in short-term HSPCs. The combinatorial use of small molecules, such as staurosporine and PGE₂, to enhance LVV transduction of human CD34⁺ cells is a promising method to improve transduction efficiency and subsequent potential therapeutic benefit of gene therapy drug products.

SUBMITTER: Lewis G 

PROVIDER: S-EPMC6054695 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature