In vivo delivery of human alpha-L-iduronidase in mice implanted with neo-organs.
Mucopolysaccharidose type I is a lysosomal storage disease caused by a deficiency in the enzyme alpha-L-iduronidase (IDUA). The existence of a secretory pathway for lysosomal enzymes and the capture of secreted molecules by distant cells through binding to mannose-6-phosphate receptors have provided a rationale for enzyme replacement therapy in lysosomal storage diseases. We have used genetically modified fibroblasts implanted into neo-organs as an in vivo delivery system for IDUA. The human IDUA cDNA was isolated and inserted into a retroviral vector where it was expressed from the phosphoglycerate kinase 1 gene promoter. MPS I fibroblasts transduced with this vector showed high levels of IDUA activity and secreted phosphorylated molecules that could be internalized by naive deficient cells. Neo-organs containing 2 x 10(7) IDUA-secreting cells were implanted into nude mice. Human and murine IDUA activities were measured in the liver and spleen of animals sacrificed 35-77 days after implantation. Human IDUA activity corresponded to 0.6-2.3% of the murine enzyme activity in the liver and to 0.1-0.3% in the spleen. These data indicated that human IDUA was secreted from neo-organs and internalized in distant tissues.
Laboratoire Rétrovirus et Transfert Génétique, CNRS URA 1157, Institut Pasteur, Paris, France.
A
A
Salvetti
P
P
Moullier
V
V
Cornet
D
D
Brooks
J J
JJ
Hopwood
O
O
Danos
J M
JM
Heard
D000818:Animals; D001483:Base Sequence; D017690:Cell Transplantation; D002478:Cells, Cultured; D018076:DNA, Complementary; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000637:transplantation* / Q000821:virology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000302:isolation & purification; D006801:Humans; D007068:Iduronidase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000737:chemistry / Q000201:enzymology; D016464:Lysosomal Storage Diseases / Q000235:genetics / Q000628:therapy; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics; D012867:Skin / Q000166:cytology; D013154:Spleen / Q000737:chemistry / Q000201:enzymology; D014161:Transduction, Genetic; D014182:Transplantation, Autologous
D018076:DNA, Complementary; D007068:Iduronidase