title: Retroviral transfer and long-term expression of the adrenoleukodystrophy gene in human CD34+ cells.
abstract: Adrenoleukodystrophy (ALD) is a demyelinating disease of the central nervous system that results from a genetic deficiency of ALDP, an ABC protein involved in the transport of very long-chain fatty acids (VLCFAs). The cloning of the ALD gene and the positive effects of allogeneic bone marrow transplantation support the feasibility of a gene therapy approach. We report the retroviral transfer of the ALD cDNA to peripheral blood and bone marrow CD34+ cells from control donors and ALD patients. Prestimulation of these cells with cytokines, followed by infection with the M48-ALD retroviral vector, resulted in 20% transduction efficiency (4-40%) and expression of the vector-encoded ALDP in 20% of CD34+ cells (7.3-50%). Long-term culture (LTC) of transduced CD34+ cells from two ALD patients showed efficient transduction (24-28%) and stable expression (25-32%) of ALDP in derived clonogenic progenitors at 3 weeks of culture. The expression of ALDP in CFU cells derived from 5 and 6 weeks of LTC confirmed the effective transduction of LTC-initiating cells. Expression of ALDP was observed in CD68+ CFU-derived cells, suggesting that monocyte-macrophages, the target bone marrow cells in ALD, were produced from transduced progenitor cells. VL-CFA content was corrected in LTC and CFU-derived cells in proportion to the percentage of transduced cells, indicating that the vector-encoded ALDP was functional. Although not efficient yet to allow a clinical perspective, these results demonstrate the feasibility of ALD gene transfer into CD34+ cells of ALD patients.
authors.affiliation: INSERM U342, Hôpital Saint-Vincent de Paul, Paris, France.
authors.forename: N
authors.initials: N
authors.lastname: Doerflinger
authors.forename: J M
authors.initials: JM
authors.lastname: Miclea
authors.forename: J
authors.initials: J
authors.lastname: Lopez
authors.forename: C
authors.initials: C
authors.lastname: Chomienne
authors.forename: P
authors.initials: P
authors.lastname: Bougnères
authors.forename: P
authors.initials: P
authors.lastname: Aubourg
authors.forename: N
authors.initials: N
authors.lastname: Cartier
mesh_terms: D016475:3T3 Cells; D000074063:ATP Binding Cassette Transporter, Subfamily D, Member 1; D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000326:Adrenoleukodystrophy / Q000235:genetics*; D000818:Animals; D018952:Antigens, CD34; D001854:Bone Marrow Cells / Q000378:metabolism*; D018929:Cell Culture Techniques; D003115:Colony-Stimulating Factors / Q000494:pharmacology; D018076:DNA, Complementary; D005227:Fatty Acids / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D012190:Retroviridae; D013997:Time Factors; D014162:Transfection
chemical_list: C115572:ABCD1 protein, human; D000074063:ATP Binding Cassette Transporter, Subfamily D, Member 1; D018528:ATP-Binding Cassette Transporters; D018952:Antigens, CD34; D003115:Colony-Stimulating Factors; D018076:DNA, Complementary; D005227:Fatty Acids; D008565:Membrane Proteins