Stable and functional lymphoid reconstitution in artemis-deficient mice following lentiviral artemis gene transfer into hematopoietic stem cells

Mol Ther. 2008 Aug;16(8):1490-9. doi: 10.1038/mt.2008.118. Epub 2008 Jun 17.

Abstract

Patients with mutations in the Artemis gene display a complete absence of T- and B lymphocytes, together with increased cellular radiosensitivity; this leads to a radiosensitive severe combined immunodeficiency (RS-SCID). Allogenic hematopoietic stem-cell (HSC) transplantation is only partially successful in the absence of an human leukocyte antigen-genoidentical donor, and this has prompted a search for alternative therapeutic approaches such as gene therapy. In this study, a self-inactivated lentiviral vector expressing Artemis was used to complement the Artemis knockout mouse (Art(-/-)). Transplantation of Artemis-transduced HSCs into irradiated Art(-/-) mice restored a stable (over a 15-month period of follow-up) and functional T- and cell repertoire that was comparable to that of control mice. The success of secondary transplantations demonstrated that the HSCs had been transduced. One of thirteen mice developed a thymoma 6 months after gene therapy. Although thymic cells were seen to be carrying two lentiviral integration sites, there was no evidence of lentivirus-driven oncogene activation. The Art(-/-) mice were found to be prone to develop T-cell lymphomas, either spontaneously or after irradiation. These data indicate that the observed lymphoproliferation was probably the consequence of the chromosomal instability associated with the Artemis-deficient background. As a whole, our work provides a basis for supporting the gene therapy approach in Artemis-deficient SCID.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes / immunology
  • CD3 Complex / analysis
  • CD4 Antigens / analysis
  • CD8 Antigens / analysis
  • Cells, Cultured
  • Endonucleases
  • Female
  • Flow Cytometry
  • Genetic Therapy / methods
  • Genetic Vectors / genetics
  • Hematopoietic Stem Cell Transplantation / methods*
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / metabolism
  • Hyaluronan Receptors / analysis
  • Interleukin-2 Receptor alpha Subunit / analysis
  • Lentivirus / genetics*
  • Male
  • Mice
  • Mice, Knockout
  • Nuclear Proteins / deficiency
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / metabolism
  • Severe Combined Immunodeficiency / genetics
  • Severe Combined Immunodeficiency / immunology
  • Severe Combined Immunodeficiency / therapy*
  • T-Lymphocytes / immunology
  • Thymus Gland / immunology
  • Thymus Gland / pathology
  • Transduction, Genetic

Substances

  • CD3 Complex
  • CD4 Antigens
  • CD8 Antigens
  • Hyaluronan Receptors
  • Interleukin-2 Receptor alpha Subunit
  • Nuclear Proteins
  • Endonucleases
  • Dclre1c protein, mouse