TY - JOUR T1 - Preferential Expansion of Human CD34CD133CD90 Hematopoietic Stem Cells Enhances Gene-Modified Cell Frequency for Gene Therapy. JF - Hum Gene Ther Y1 - 2022 A1 - Christopher, Abisha Crystal A1 - Venkatesan, Vigneshwaran A1 - Karuppusamy, Karthik V A1 - Srinivasan, Saranya A1 - Babu, Prathibha A1 - Azhagiri, Manoj Kumar K A1 - Chambayil, Karthik A1 - Bagchi, Abhirup A1 - Rajendiran, Vignesh A1 - Ravi, Nithin Sam A1 - Kumar, Sanjay A1 - Marepally, Srujan Kumar A1 - Mohankumar, Kumarasamypet Murugesan A1 - Srivastava, Alok A1 - Velayudhan, Shaji R A1 - Thangavel, Saravanabhavan KW - Animals KW - Antigens, CD34 KW - Fetal Blood KW - Genetic Therapy KW - Hematopoietic Stem Cell Transplantation KW - Hematopoietic Stem Cells KW - Humans KW - Mice KW - Mice, Inbred NOD KW - Mice, SCID AB -

CD34CD133CD90 hematopoietic stem cells (HSCs) are responsible for long-term multilineage hematopoiesis, and the high frequency of gene-modified HSCs is crucial for the success of hematopoietic stem and progenitor cell (HSPC) gene therapy. However, the culture and gene manipulation steps of HSPC graft preparation significantly reduce the frequency of HSCs, thus necessitating large doses of HSPCs and reagents for the manipulation. In this study, we identified a combination of small molecules, Resveratrol, UM729, and SR1 that preferentially expands CD34CD133CD90 HSCs over other subpopulations of adult HSPCs in culture. The preferential expansion enriches the HSCs in culture, enhances the adhesion, and results in a sixfold increase in the long-term engraftment in NSG mice. Further, the culture-enriched HSCs are more responsive to gene modification by lentiviral transduction and gene editing, increasing the frequency of gene-modified HSCs up to 10-fold . The yield of gene-modified HSCs obtained by the culture enrichment is similar to the sort-purification of HSCs and superior to Cyclosporin-H treatment. Our study addresses a critical challenge of low frequency of gene modified HSCs in HSPC graft by developing and demonstrating a facile HSPC culture condition that increases the frequency of gene-modified cells . This strategy will improve the outcome of HSPC gene therapy and also simplify the gene manipulation process.

VL - 33 IS - 3-4 ER -