Long-term expression and secretion of human glucocerebrosidase by primary murine and human myoblasts and differentiated myotubes.
Gaucher disease (GD) is caused by a deficiency in glucocerebrosidase (GC). Enzyme replacement for GD disease is effective but expensive and requires life-long treatment. Development of alternative therapeutic strategies is therefore important. One approach is an enzyme delivery system which could supply GC into the circulation continuously. We have previously reported that human GC cDNA in a retroviral vector (MFG-GC) efficiently transduced a murine myoblast line (C2C12) and expressed GC intracellularly and extracellularly. Now we have demonstrated that primary murine and human myoblasts are transduced at very high efficiency by MFG-GC (five to ten copies of human GC gene per cell at a multiplicity of infection of 5-10), 100% of MFG-GC transduced cells expressed human GC. The transduced primary murine and human myoblasts had an intracellular GC activities about five to ten times above nontransduced controls. Furthermore, transduced primary myoblasts secreted human GC extracellularly for up to 35 weeks in vitro. The secreted human GC is specifically taken up by bone marrow derived macrophages, the cell type most important to the pathogenesis of GD. These data suggest that transduced primary myoblasts may be useful in supplying GC as an alternative approach to the treatment of GD.
Liu
C
C
Department of Human Genetics, Graduate School of Public Health, University of Pittsburgh, PA 15261, USA.
Bahnson
A B
AB
Dunigan
J T
JT
Watkins
S C
SC
Barranger
J A
JA
D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D002454:Cell Differentiation; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D018920:Coculture Techniques; D005110:Extracellular Space; D005260:Female; D015870:Gene Expression; D005962:Glucosylceramidase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007424:Intracellular Fluid; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D018482:Muscle, Skeletal / Q000166:cytology*; D013997:Time Factors
D005962:Glucosylceramidase