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Efficient retrovirus-mediated transfer of the multidrug resistance 1 gene into autologous human long-term repopulating hematopoietic stem cells

Nature Medicine volume 6, pages 652–658 (2000)Cite this article

Abstract

Pre-clinical studies indicate that efficient retrovirus-mediated gene transfer into hematopoietic stem cells and progenitor cells can be achieved by co-localizing retroviral particles and target cells on specific adhesion domains of fibronectin. In this pilot study, we used this technique to transfer the human multidrug resistance 1 gene into stem and progenitor cells of patients with germ cell tumors undergoing autologous transplantation. There was efficient gene transfer into stem and progenitor cells in the presence of recombinant fibronectin fragment CH-296. The infusion of these cells was associated with no harmful effects and led to prompt hematopoietic recovery. There was in vivo vector expression, but it may have been limited by the high rate of aberrant splicing of the multidrug resistance 1 gene in the vector. Gene marking has persisted more than a year at levels higher than previously reported in humans.

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Figure 1: Treatment protocol.
Figure 2: Transgene frequency in peripheral blood leukocytes as determined by quantitative real-time PCR.
Figure 3: Transgene integrity.

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Acknowledgements

We thank the patients and their families for their contributions to this study. We thank the Indiana University General Clinical Research Center, Stem Cell Laboratory (V. Graves and J. Baily), Apheresis, and BMT unit staff for excellent technical and clinical care. We thank S. Harker from the General Clinical Research Center nursing staff for her contribution to this study. We thank the members of the Gene Therapy Working Group for their contribution (L. Rubin, J. Croop, M. Dinauer, S. Richeson, L. Reeves, J. Good and S. Holbrook). This work was supported by the National Centers for Research Resources (National Institutes of Health M01 RR00750) (R.A.) and an American Cancer Society grant CRTG-97-042-EDT (R.A.). The National Gene Vector Laboratory (U42 RR/1148 and CA11148) produced the clinical-grade vector. Additional support was provided by the Indiana University Vector Core and the Stem Cell Laboratory Core (2 P30 DK49218 and P01 HL 53586) (K.C. and R.A.). Takara Shuzo (Otsu, Japan) has provided the Gene Therapy Working Group with an unrestricted grant to help conduct clinical gene therapy trials at our center. A.B. has an equity interest in Genetix Pharmaceuticals, which provided the vector-packaging cell line (A12M1) to the National Gene Vector Laboratory for this study. D.A.W. may receive royalties based on ‘milestones’ set forth in a licensing agreement between Takara Shuzo and Indiana University.

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Authors and Affiliations

  1. Department of Medicine, Howard Hughes Medical Institute Indiana University School of Medicine, 1044 W. Walnut, Rm. 402 , Indianapolis, 46202,, IN, USA

    Rafat Abonour, Lawrence Einhorn, Christie M. Traycoff, Stephen D. Williams, Robert Hromas, Michael J. Robertson, Franklin O. Smith, Edward F. Srour & Kenneth Cornetta

  2. Department of Microbiology and Immunology, Howard Hughes Medical Institute, Indiana University School of Medicine, 1044 W. Walnut, Rm. 402,, Indianapolis, 46202 , IN, USA

    David A. Williams, Robert Hromas, Franklin O. Smith, Edward F. Srour & Kenneth Cornetta

  3. Department of Pediatrics, Howard Hughes Medical Institute Indiana University School of Medicine, 1044 W. Walnut, Rm. 402 , Indianapolis, 46202, IN, USA

    David A. Williams & Franklin O. Smith

  4. Herman B Wells Center for Pediatrics Research, Howard Hughes Medical Institute, Indiana University School of Medicine, 1044 W. Walnut, Rm. 402, Indianapolis, 46202, IN, USA

    David A. Williams

  5. Columbia University, 701st West 168th St., HHSC16-1604, New York, 10032, New York, USA

    Kristin M. Hall, Jun Chen, John Coffman, Arthur Bank & Maureen Ward

  6. Takara Shuzo, Biomedical Group, Seta 3-4-1, Otsu, 520-2193 , Shiga, Japan

    Ikunoshin Kato

  7. Department of Medical and Molecular Genetics, Indiana University School of Medicine, 1044 W. Walnut, Rm. 202, Indianapolis, 46202, Indiana, USA

    Robert Hromas & Kenneth Cornetta

  8. AMGEN, One Amgen Center Drive, Thousand Oaks, 91320-1799, California, USA

    David Woo

  9. Nexell Therapeutics, 9 Parker, Irvine, 92618, California, USA

    Bonnie Mills

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  1. Rafat Abonour
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Abonour, R., Williams, D., Einhorn, L. et al. Efficient retrovirus-mediated transfer of the multidrug resistance 1 gene into autologous human long-term repopulating hematopoietic stem cells . Nat Med 6, 652–658 (2000). https://doi.org/10.1038/76225

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