Long-term in vitro correction of alpha-L-iduronidase deficiency (Hurler syndrome) in human bone marrow.

Abstract

Allogeneic bone marrow transplantation is the most effective treatment for Hurler syndrome but, since this therapy is not available to all patients, we have considered an alternative approach based on transfer and expression of the normal gene in autologous bone marrow. A retroviral vector carrying the full-length cDNA for a-L-iduronidase has been constructed and used to transduce bone marrow from patients with this disorder. Various gene-transfer protocols have been assessed including the effect of intensive schedules of exposure of bone marrow to viral supernatant and the influence of growth factors. With these protocols, we have demonstrated successful gene transfer into primitive CD34+ cells and subsequent enzyme expression in their maturing progeny. Also, by using long-term bone marrow cultures, we have demonstrated high levels of enzyme expression sustained for several months. The efficiency of gene transfer has been assessed by PCR analysis of hemopoietic colonies as 25-56%.

Correction in trans for Fabry disease: expression, secretion and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector.

“…Along these lines, lymphocytes from patients with Hunter syndrome were corrected endogenously for the enzymatic defect by retroviral-mediated gene transfer and cell-tocell enzyme activity transmission occurred (23). Another recent study demonstrated sustained long-term expression and secretion of a-L-iduronidase, another lysosomal enzyme, in transduced human stem/progenitor cells (24). This further demonstrates the potential for metabolic cooperativity for many diseases of this type.…”

Retrovirally mediated correction of bone marrow–derived mesenchymal stem cells from patients with mucopolysaccharidosis type I

“…17 Lenhanced green fluorescent protein (L-EGFP) was derived by replacing the IDUA complementary DNA in Lid with a complementary DNA encoding L-EGFP. Cells (30%-40% confluent) were transduced using cell-free retroviral supernatant supplemented with 2 g/mL polybrene.…”

Chemoprotective gene transfer I: transduction of human haemopoietic progenitors with O6-benzylguanine-resistant O6-alkylguanine-DNA alkyltransferase attenuates the toxic effects of O6-alkylating agents in vitro

“…21 The D 37 values obtained for temozolomide and mitozolomide, as well as for another chloroethylating agent, chlorozotocin (Table 2), demonstrated that the retroviral delivery and expression of hATPA/GA can significantly protect human haemopoietic cells against the toxicity of a wide + cells were transduced with LhATPA/GA, exposed to increasing temozolomide concentrations in the presence (10 m) or absence of O 6 -beG and the survival of the GM-CFC determined ( Figure 2a). As controls, cells were transduced with a retrovirus harbouring the ␣-iduronidase cDNA (LID) 20 and were treated identically and in parallel with LhATPA/GAtransduced cells. Under conditions in which a transduction efficiency of 70% was achieved, as determined by PCR, CD34…”