title: Adenovirus-mediated correction of the genetic defect in hepatocytes from patients with familial hypercholesterolemia.
abstract: Familial hypercholesterolemia (FH) is an inherited deficiency of LDL receptors that has been an important model for liver-directed gene therapy. We are developing approaches for treating FH that are based on direct delivery of recombinant LDL receptor genes to liver in vivo. As a first step towards this goal, replication-defective recombinant adenoviruses were constructed which contained either the lacZ gene or the human LDL receptor cDNA expressed from a beta-actin promoter. Primary cultures of hepatocytes were established from two patients with homozygous FH and one nonFH patient, and subsequently exposed to recombinant adenoviruses at MOIs ranging from 0.1 to 5. Essentially all of the cells expressed high levels of the transgene without demonstrable expression of an early or late adenoviral gene product; the level of recombinant-derived LDL receptor protein in transduced FH hepatocytes exceeded the endogenous levels by at least 20-fold. These studies support the utility of recombinant adenoviruses for efficient transduction of recombinant LDL receptor genes into human FH hepatocytes without expression of viral proteins.
authors.affiliation: Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor 48109.
authors.forename: K
authors.initials: K
authors.lastname: Kozarsky
authors.forename: M
authors.initials: M
authors.lastname: Grossman
authors.forename: J M
authors.initials: JM
authors.lastname: Wilson
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000328:Adult; D002478:Cells, Cultured; D002648:Child; D003673:Defective Viruses / Q000235:genetics; D005260:Female; D005816:Genetic Complementation Test; D015316:Genetic Therapy / Q000379:methods*; D006720:Homozygote; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000235:genetics* / Q000628:therapy; D008099:Liver / Q000166:cytology / Q000502:physiology*; D008297:Male; D011973:Receptors, LDL / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D014161:Transduction, Genetic
chemical_list: D011973:Receptors, LDL; D011994:Recombinant Proteins