Retroviral-mediated transfer of the human glucocerebrosidase gene into cultured gaucher bone marrow

Jan Nolta, Xiao Jin Yu, Ingrid Bahner, Donald B. Kohn

Research output: Contribution to journalArticlepeer-review

63 Scopus citations


Gaucher disease, a lysosomal glycolipid storage disorder, results from the genetic deficiency of an acidic glucosidase, glucocerebrosidase (GC). The beneficial effects of allogeneic bone marrow transplantation (BMT) for Gaucher disease suggest that GC gene transduction and the transplantation of autologous hematopoietic stem cells (gene therapy) may similarly alleviate symptoms. We have constructed a retroviral vector, L-GC, produced by a clone of the amphotropic packaging cell line PA317, which transduces the normal human GC cDNA with high efficiency. Whole-marrow mononuclear cells and CD34-enriched cells from a 4-yr-old female with type 3 Gaucher disease were transduced by the L-GC vector and studied in long-term bone marrow culture (LTBMC). Prestimulation of marrow with IL-3 and IL-6, followed by co-cultivation with vector-producing fibroblasts, produced gene transfer into 40-45% of the hematopoietic progenitor cells. The levels of GC expression in progeny cells (primarily mature myelomonocytic) produced by the LTBMC were quantitatively analyzed by Northern blot, Western blot, and glucocerebrosidase enzyme assay. Normal levels of GC RNA, immunoreactive protein, and enzymatic activity were detected throughout the duration of culture. These studies demonstrate that retroviral vectors can efficiently transfer the GC gene into long-lived hematopoietic progenitor cells from the bone marrow of patients with Gaucher disease and express physiologically relevant levels of GC enzyme activity.

Original languageEnglish (US)
Pages (from-to)342-348
Number of pages7
JournalJournal of Clinical Investigation
Issue number2
StatePublished - 1992
Externally publishedYes


  • Gaucher disease
  • Gene therapy
  • Long-term bone marrow culture
  • Long-term culture-initiating cells

ASJC Scopus subject areas

  • Medicine(all)


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