In utero injection of alpha-L-iduronidase-carrying retrovirus in canine mucopolysaccharidosis type I: infection of multiple tissues and neonatal gene expression.
Canine alpha-L-iduronidase (alpha-ID) deficiency is caused by a single base pair mutation in the alpha-ID gene, resulting in no enzyme activity in homozygous affected pups. The disease clinically resembles human mucopolysaccharidosis type I (MPSI). We used the canine MPSI model system to address the efficacy of a new retroviral vector, MND-MFG, containing the human alpha-ID cDNA (MND-MFG-alpha-ID) for direct in utero gene delivery to MPSI cells. In vitro, the MND-MFG-alpha-ID vector showed high-level, long-term expression of the transgene in both canine and human alpha-ID-deficient fibroblasts. The effectiveness of this vector for in utero gene transfer and expression in multiple tissues was assessed by injecting viral supernatants into MPSI fetuses and evaluating transduction efficiency and enzyme expression at various times after birth. Transduction of a spectrum of cell types and tissues was observed in all seven live-born pups and in one stillborn pup. Although enzyme activity was not detected in adult tissues from the seven surviving pups, significant alpha-ID enzyme activity was detected in both the liver and kidney of the deceased pup. Our combined gene delivery vector and in utero transfer approach, while encouraging in terms of overall gene transfer efficiency to multiple tissues and successful short-term gene expression, was unable to meet the important requirement of sustained in vivo gene expression.
Meertens
Lisa
L
Department of Clinical Pathology, Sunnybrook and Women's College Health Science Centre, Toronto, ON M4N 3M5, Canada.
Zhao
Yongjun
Y
Rosic-Kablar
Suzana
S
Li
Liheng
L
Chan
Kin
K
Dobson
Howard
H
Gartley
Cathy
C
Lutzko
Carolyn
C
Hopwood
John
J
Kohn
Donald
D
Kruth
Stephen
S
Hough
Margaret R
MR
Dubé
Ian D
ID
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use; D004195:Disease Models, Animal; D004285:Dogs; D005260:Female; D005315:Fetal Diseases / Q000201:enzymology / Q000628:therapy*; D005347:Fibroblasts / Q000201:enzymology; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000627:therapeutic use; D006801:Humans; D007068:Iduronidase / Q000172:deficiency / Q000235:genetics*; D007267:Injections; D007274:Injections, Intraperitoneal; D019169:Jurkat Cells / Q000201:enzymology; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008059:Mucopolysaccharidosis I / Q000196:embryology / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D011247:Pregnancy; D014018:Tissue Distribution; D014161:Transduction, Genetic; D015017:Yolk Sac
D018076:DNA, Complementary; D007068:Iduronidase