TRANSDUCTION OF CD34-BLOOD AND GAUCHER BONE-MARROW CELLS BY A RETROVIRAL VECTOR CARRYING THE GLUCOCEREBROSIDASE GENE( ENRICHED CORD)

One promising strategy for gene therapy of Gaucher disease involves ex vivo retroviral transduction of autologous hematopoietic stem cells. Studies in small animals have demonstrated that this approach provides a life-long supply of the glucocerebrosidase (GC) enzyme. Human appli cation has developed to the stage of a clinical trial. In this study, we describe development of a high titer amphotropic producer line for the vector, MFG-GC, and explore transduction of CD34+ cells from vario us human sources. Higher than three times the normal levels of glucoce rebrosidase activity in non-transduced cells were achieved following t ransduction of CD34+ cells obtained from bone marrow or cord blood fro m normal donors. The improvement in enzyme activity in Gaucher marrow was about 40-fold above deficient levels. We examined the timing and s tepwise effect of multiple rounds of infection and evaluated post-infe ction expansion of cells in two different cytokine mixtures. Transduct ion efficiency was determined using immunocytochemistry and Southern b lot hybridization.