title: Reconstitution of enzymatic activity in hepatocytes of phenylalanine hydroxylase-deficient mice.
abstract: Phenylketonuria (PKU) is a metabolic disorder secondary to a deficiency of the hepatic enzyme phenylalanine hydroxylase (PAH). The recent creation of a mouse strain for PAH deficiency has provided an excellent model system to explore the possibility of its phenotypic correction by hepatic gene therapy. A recombinant retrovirus containing the mouse PAH cDNA under the transcriptional control of the human CMV promoter was constructed and used to transduce hepatocytes isolated from PAH-deficient mice. Viral-transduced hepatocytes produced dramatically higher levels of mouse PAH mRNA as compared to control mock-infected hepatocytes. The PAH mRNA was translated efficiently into PAH protein that is capable of converting phenylalanine to tyrosine in vitro. These results demonstrate that the PAH-deficient mouse hepatocytes can be readily reconstituted by retroviral-mediated gene transduction, which is a crucial step towards somatic gene therapy for PKU.
authors.affiliation: Howard Hughes Medical Institute, Department of Cell Biology, Houston, Texas 77030.
authors.forename: T J
authors.initials: TJ
authors.lastname: Liu
authors.forename: M A
authors.initials: MA
authors.lastname: Kay
authors.forename: G J
authors.initials: GJ
authors.lastname: Darlington
authors.forename: S L
authors.initials: SL
authors.lastname: Woo
mesh_terms: D000818:Animals; D005822:Genetic Vectors; D008099:Liver / Q000737:chemistry / Q000166:cytology / Q000201:enzymology*; D051379:Mice; D008817:Mice, Mutant Strains; D010651:Phenylalanine Hydroxylase / Q000172:deficiency* / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism; D012192:Retroviridae Infections / Q000201:enzymology / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D011994:Recombinant Proteins; D010651:Phenylalanine Hydroxylase