A Combined In Vivo HSC Transduction/Selection Approach Results in Efficient and Stable Gene Expression in Peripheral Blood Cells in Mice

Hongjie Wang, Maximilian Richter, Nikoletta Psatha, Chang Li, Jiho Kim, Jing Liu, Anja Ehrhardt, Susan K. Nilsson, Benjamin Cao, Donna Palmer, Philip Ng, Zsuzsanna Izsvák, Kevin G. Haworth, Hans Peter Kiem, Thalia Papayannopoulou, André Lieber

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14 Citations (Scopus)

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 O6-methylguanine-DNA methyltransferase (mgmtP140K) gene that confers resistance to O6-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.

Original languageEnglish
Pages (from-to)52-64
Number of pages13
JournalMolecular Therapy - Methods & Clinical Development
Volume8
DOIs
Publication statusPublished - 16 Mar 2018

Keywords

  • adenovirus
  • hematopoietic stem cells
  • mobilization

Cite this

Wang, H., Richter, M., Psatha, N., Li, C., Kim, J., Liu, J., Ehrhardt, A., Nilsson, S. K., Cao, B., Palmer, D., Ng, P., Izsvák, Z., Haworth, K. G., Kiem, H. P., Papayannopoulou, T., & Lieber, A. (2018). A Combined In Vivo HSC Transduction/Selection Approach Results in Efficient and Stable Gene Expression in Peripheral Blood Cells in Mice. Molecular Therapy - Methods & Clinical Development, 8, 52-64. https://doi.org/10.1016/j.omtm.2017.11.004