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A combined in vivo HSC transduction/selection approach results in efficient and stable gene expression in peripheral blood cells in mice

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Item Type:Article
Title:A combined in vivo HSC transduction/selection approach results in efficient and stable gene expression in peripheral blood cells in mice
Creators Name:Wang, H. and Richter, M. and Psatha, N. and Li, C. and Kim, J. and Liu, J. and Ehrhardt, A. and Nilsson, S.K. and Cao, B. and Palmer, D. and Ng, P. and Izsvák, Z. and Haworth, K.G. and Kiem, H.P. and Papayannopoulou, T. and Lieber, A.
Abstract:We recently reported on an in vivo hematopoietic stem cell (HSC) gene therapy approach. It involves the subcutaneous injections of G-CSF/AMD3100 to mobilize HSCs from the bone marrow into the peripheral blood stream and the intravenous injection of an integrating helper-dependent adenovirus vector system. HSCs transduced in the periphery homed back to the bone marrow, where they persisted long-term. However, high transgene marking rates found in primitive bone marrow HSCs were not reflected in peripheral blood cells. Here, we tested small-molecule drugs to achieve selective mobilization and transduction of HSCs. We found more efficient GFP marking in bone marrow HSCs but no increased marking in the peripheral blood cells. We then used an in vivo HSC chemo-selection based on a mutant of the O(6)-methylguanine-DNA methyltransferase (mgmt(P140K)) gene that confers resistance to O(6)-BG/BCNU and should give stably transduced HSCs a proliferation stimulus and allow for the selective survival and expansion of progeny cells. Short-term exposure of G-CSF/AMD3100-mobilized, in vivo-transduced mice to relatively low selection drug doses resulted in stable GFP expression in up to 80% of peripheral blood cells. Overall, the further improvement of our in vivo HSC transduction approach creates the basis for a simpler HSC gene therapy.
Keywords:Hematopoietic Stem Cells, Adenovirus, Mobilization, Animals, Mice
Source:Molecular Therapy - Methods and Clinical Development
ISSN:2329-0501
Publisher:Cell Press (U.K)
Volume:8
Page Range:52-64
Date:16 March 2018
Official Publication:https://doi.org/10.1016/j.omtm.2017.11.004
PubMed:View item in PubMed

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