Transduction of hepatocytes after neonatal delivery of a Moloney murine leukemia virus based retroviral vector results in long-term expression of beta-glucuronidase in mucopolysaccharidosis VII dogs.
The use of Moloney murine leukemia virus (MLV)-based retroviral vectors (RV) can result in stable in vivo expression in the liver, but these vectors only transduce replicating hepatocytes. As newborn animals exhibit rapid growth, we evaluated the ability of MLV-based RV to transduce hepatocytes in neonatal dogs. I.v. injection of a beta-galactosidase-expressing RV at 3 days after birth resulted in transduction of 9% of hepatocytes. Prior treatment with human hepatocyte growth factor at 2.5 mg/kg did not increase transduction. Although cells from the spleen were also transduced with moderate efficiency, cells from other organs were not. Neonatal dogs with mucopolysaccharidosis VII (MPS VII) received an i.v.injection of an RV containing the canine beta-glucuronidase (cGUSB) cDNA. At several months after transduction, clusters of hepatocytes that expressed high levels of cGUSB were present in the liver, which probably derived from replication of transduced hepatocytes. At 6 months after transduction, serum GUSB levels were 73% that of homozygous normal dogs and were 34% of the peak values observed at 1 week. We conclude that neonatal delivery of an MLV-based RV results in stable transduction of hepatocytes in dogs. This approach could result in immediate correction in patients with an otherwise-lethal genetic deficiency.
Xu
Lingfei
L
Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Haskins
Mark E
ME
Melniczek
John R
JR
Gao
Cuihua
C
Weil
Margaret A
MA
O'Malley
Thomas M
TM
O'Donnell
Patricia A
PA
Mazrier
Hamutal
H
Ellinwood
N Matthew
NM
Zweigle
Jean
J
Wolfe
John H
JH
Ponder
Katherine Parker
KP
D000818:Animals; D000831:Animals, Newborn; D001973:Bromodeoxyuridine; D002455:Cell Division; D004285:Dogs; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005966:Glucuronidase / Q000235:genetics; D017228:Hepatocyte Growth Factor; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008979:Moloney murine leukemia virus / Q000502:physiology*; D016538:Mucopolysaccharidosis VII / Q000378:metabolism* / Q000473:pathology / Q000628:therapy; D013154:Spleen / Q000166:cytology; D014161:Transduction, Genetic
D017228:Hepatocyte Growth Factor; D005966:Glucuronidase; D001973:Bromodeoxyuridine