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Integrating HDAd5/35++ vectors as a new platform for HSC gene therapy of hemoglobinopathies

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Item Type:Article
Title:Integrating HDAd5/35++ vectors as a new platform for HSC gene therapy of hemoglobinopathies
Creators Name:Li, C., Psatha, N., Wang, H., Singh, M., Samal, H.B., Zhang, W., Ehrhardt, A., Izsvák, Z., Papayannopoulou, T. and Lieber, A.
Abstract:We generated an integrating, CD46-targeted, helper-dependent adenovirus HDAd5/35++ vector system for hematopoietic stem cell (HSC) gene therapy. The ∼12-kb transgene cassette included a β-globin locus control region (LCR)/promoter driven human γ-globin gene and an elongation factor alpha-1 (EF1α)-mgmtP140K expression cassette, which allows for drug-controlled increase of γ-globin-expressing erythrocytes. We transduced bone marrow lineage-depleted cells from human CD46-transgenic mice and transplanted them into lethally irradiated recipients. The percentage of γ-globin-positive cells in peripheral blood erythrocytes in primary and secondary transplant recipients was stable and greater than 90%. The γ-globin level was 10%–20% of adult mouse globin. Transgene integration, mediated by a hyperactive Sleeping Beauty SB100x transposase, was random, without a preference for genes. A second set of studies was performed with peripheral blood CD34+ cells from mobilized donors. 10 weeks after transplantation of transduced cells, human cells were harvested from the bone marrow and differentiated ex vivo into erythroid cells. Erythroid cells expressed γ-globin at a level of 20% of adult α-globin. Our studies suggest that HDAd35++ vectors allow for efficient transduction of long-term repopulating HSCs and high-level, almost pancellular γ-globin expression in erythrocytes. Furthermore, our HDAd5/35++ vectors have a larger insert capacity and a safer integration pattern than currently used lentivirus vectors.
Keywords:Hematopoietic Stem Cells, Sleeping Beauty Transposase, beta-Thalassemia, Sickle Cell Disease, gamma Globin, Integration, Gene Addition, Animals, Mice
Source:Molecular Therapy - Methods and Clinical Development
Publisher:Cell Press
Page Range:142-152
Date:15 June 2018
Official Publication:https://doi.org/10.1016/j.omtm.2018.02.004
PubMed:View item in PubMed

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