CD133-targeted Gene Transfer Into Long-term Repopulating Hematopoietic Stem Cells
Authors: Christian Brendel, Benjamin Goebel, Abriss Daniela, Martijn Brugman, Sabrina Kneissl, Joachim Schwäble, Kerstin B Kaufmann, Uta Müller-Kuller, Hana Kunkel, Linping Chen-Wichmann, Tobias Abel, Hubert Serve, Leonid Bystrykh, Christian J Buchholz, Manuel Grez
Journal: Molecular Therapy (2014)

Gene therapy for hematological disorders relies on the genetic modification of CD34+ cells, a heterogeneous cell population containing about 0.01% long-term repopulating cells. Here, we show that the lentiviral vector CD133-LV, which uses a surface marker on human primitive hematopoietic stem cells (HSCs) as entry receptor, transfers genes preferentially into cells with high engraftment capability. Transduction of unstimulated CD34+ cells with CD133-LV resulted in gene marking of cells with competitive proliferative advantage in vitro and in immunodeficient mice. The CD133-LV-transduced population contained significantly more cells with repopulating capacity than cells transduced with vesicular stomatitis virus (VSV)-LV, a lentiviral vector pseudotyped with the vesicular stomatitis virus G protein. Upon transfer of a barcode library, CD133-LV-transduced cells sustained gene marking in vivo for a prolonged period of time with a 6.7-fold higher recovery of barcodes compared to transduced control cells. Moreover, CD133-LV-transduced cells were capable of repopulating secondary recipients. Lastly, we show that this targeting strategy can be used for transfer of a therapeutic gene into CD34+ cells obtained from patients suffering of X-linked chronic granulomatous disease. In conclusion, direct gene transfer into CD133+ cells allows for sustained long-term engraftment of gene corrected cells. Gene therapy for hematological disorders relies on the genetic modification of CD34+ cells, a heterogeneous cell population containing about 0.01% long-term repopulating cells. Here, we show that the lentiviral vector CD133-LV, which uses a surface marker on human primitive hematopoietic stem cells (HSCs) as entry receptor, transfers genes preferentially into cells with high engraftment capability. Transduction of unstimulated CD34+ cells with CD133-LV resulted in gene marking of cells with competitive proliferative advantage in vitro and in immunodeficient mice. The CD133-LV-transduced population contained significantly more cells with repopulating capacity than cells transduced with vesicular stomatitis virus (VSV)-LV, a lentiviral vector pseudotyped with the vesicular stomatitis virus G protein. Upon transfer of a barcode library, CD133-LV-transduced cells sustained gene marking in vivo for a prolonged period of time with a 6.7-fold higher recovery of barcodes compared to transduced control cells. Moreover, CD133-LV-transduced cells were capable of repopulating secondary recipients. Lastly, we show that this targeting strategy can be used for transfer of a therapeutic gene into CD34+ cells obtained from patients suffering of X-linked chronic granulomatous disease. In conclusion, direct gene transfer into CD133+ cells allows for sustained long-term engraftment of gene corrected cells.