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Lentivirus-mediated gene transfer into hematopoietic repopulating cells in baboons

Gene Therapy volume 9, pages 1464–1471 (2002)Cite this article

Abstract

Efficient transduction of hematopoietic stem cells is a prerequisite for successful hematopoietic stem cell gene therapy. Oncoretroviral vectors are the most widely used vectors for hematopoietic gene therapy studies. However, these vectors require cell division, and thus efficient transduction of quiescent stem cells has been difficult to achieve. Lentiviral vectors can transduce non-dividing cells and therefore may be more efficient in transducing quiescent hematopoietic stem cells. We have used a competitive repopulation assay in the baboon to compare transduction of hematopoietic repopulating cells by lentiviral and oncoretroviral vectors. Baboon CD34-enriched marrow cells were transduced in the presence or absence of multiple hematopoietic growth factors using a short, 2-day, transduction protocol. Here, we show that efficient lentiviral transduction of hematopoietic repopulating cells was only achieved when cells were transduced in the presence of multiple growth factors. Using these conditions, up to 8.6% of hematopoietic repopulating cells were genetically modified by the lentiviral vector more than 1 year after transplant. Interestingly, the number of lentivirally marked cells increased over time in three of four animals. In conclusion, these results suggest that lentiviral vectors are able to tranduce multilineage hematopoietic stem cells, and thus, may provide an alternative vector system for clinical stem cell gene therapy applications.

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Acknowledgements

The authors wish to thank Melissa C Richman, Laura J Peterson, and Jennifer Potter for their technical assistance, Mike Gough and the staff of the University of Washington Regional Primate Research Center for assistance with the animals, as well as Bonnie Larson and Helen Crawford for their help in preparing the manuscript. This work was supported in part by NIH grants HL54881, HL53750, DK47754, DK56465, and RR00166. PAH is supported by the German Krebshilfe. H-PK is a Markey Molecular Medicine Investigator.

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Authors and Affiliations

  1. Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    P A Horn, J C Morris, A A Bukovsky, R G Andrews, P Kurre & H-P Kiem

  2. Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA

    R G Andrews & P Kurre

  3. Department of Medicine, University of Washington School of Medicine, Seattle, WA, USA

    H-P Kiem

  4. Washington Regional Primate Research Center, University of Washington, Seattle, WA, USA

    R G Andrews & H-P Kiem

  5. Institute for Cancer Research and Treatment, University of Torino, Italy

    L Naldini

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Horn, P., Morris, J., Bukovsky, A. et al. Lentivirus-mediated gene transfer into hematopoietic repopulating cells in baboons. Gene Ther 9, 1464–1471 (2002). https://doi.org/10.1038/sj.gt.3301820

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