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Potential genotoxicity from integration sites in CLAD dogs treated successfully with gammaretroviral vector-mediated gene therapy

Gene Therapy volume 15, pages 1067–1071 (2008)Cite this article

Abstract

Integration site analysis was performed on six dogs with canine leukocyte adhesion deficiency (CLAD) that survived greater than 1 year after infusion of autologous CD34+ bone marrow cells transduced with a gammaretroviral vector expressing canine CD18. A total of 387 retroviral insertion sites (RIS) were identified in the peripheral blood leukocytes from the six dogs at 1 year postinfusion. A total of 129 RIS were identified in CD3+ T-lymphocytes and 102 RIS in neutrophils from two dogs at 3 years postinfusion. RIS occurred preferentially within 30 kb of transcription start sites, including 40 near oncogenes and 52 near genes active in hematopoietic stem cells. Integrations clustered around common insertion sites more frequently than random. Despite potential genotoxicity from RIS, to date there has been no progression to oligoclonal hematopoiesis and no evidence that vector integration sites influenced cell survival or proliferation. Continued follow-up in disease-specific animal models such as CLAD will be required to provide an accurate estimate of the genotoxicity using gammaretroviral vectors for hematopoietic stem cell gene therapy.

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Acknowledgements

We thank Dr William Telford and Veena Kapoor for assistance with flow cytometry. This research was supported by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research. Presented in part in abstract form at the 47th and 48th annual meetings of the American Society of Hematology, Atlanta, GA, December 12, 2005 and Orlando, FL, December 9, 2006, and at the 9th and 10th annual meetings of the American Society of Gene Therapy, Baltimore, MD, May 31, 2006 and Seattle, WA, May 30, 2007.

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Author notes

  1. M Hai, R L Adler and T R Bauer: These authors contributed equally to this work.

Authors and Affiliations

  1. Experimental Transplantation and Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA

    M Hai, R L Adler, T R Bauer Jr, L M Tuschong, Y-C Gu & D D Hickstein

  2. Laboratory of Molecular Technology, Scientific Applications International Corporation-Frederick, National Cancer Institute-Frederick, Frederick, Maryland, USA

    X Wu

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  1. M Hai
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  2. R L Adler
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  3. T R Bauer Jr
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  4. L M Tuschong
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  5. Y-C Gu
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  6. X Wu
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  7. D D Hickstein
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Correspondence to D D Hickstein.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Hai, M., Adler, R., Bauer, T. et al. Potential genotoxicity from integration sites in CLAD dogs treated successfully with gammaretroviral vector-mediated gene therapy. Gene Ther 15, 1067–1071 (2008). https://doi.org/10.1038/gt.2008.52

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