title: Gut epithelial cells as targets for gene therapy of hemophilia.
abstract: Gut epithelium is an attractive target for gene therapy of hemophilia due to the large number of rapidly dividing cells that should be readily accessible to a wide range of vectors by a noninvasive route of administration. We have performed in vitro tests to determine the suitability of gut epithelial cells for gene transfer, protein synthesis, and secretion of coagulation factors VIII and IX. The results with retroviral vectors indicate that transduced epithelial cells from human, rat, or porcine small or large intestine can synthesize significant amounts of factor VIII or factor IX and that two-thirds or more of the recombinant protein is secreted in a basolateral direction (i.e., away from the lumen and toward underlying capillaries and lymphatics). Furthermore, we have demonstrated that intestinal epithelial cells are susceptible to efficient gene transfer by lipofection and adenovirus vectors. In the case of factor IX, we have produced a high-titer adenovirus vector capable of transducing gut epithelial cells resulting in synthesis of factor IX. The results of our in vitro studies indicate that gene transfer targeting gut epithelium as a new approach to hemophilia gene therapy is rational and merits in vivo studies in hemophilia animal models.
authors.affiliation: Clinical Gene Therapy Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892-1851, USA.
authors.forename: J N
authors.initials: JN
authors.lastname: Lozier
authors.forename: J R
authors.initials: JR
authors.lastname: Yankaskas
authors.forename: W J
authors.initials: WJ
authors.lastname: Ramsey
authors.forename: L
authors.initials: L
authors.lastname: Chen
authors.forename: H
authors.initials: H
authors.lastname: Berschneider
authors.forename: R A
authors.initials: RA
authors.lastname: Morgan
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004847:Epithelial Cells; D005164:Factor IX / Q000235:genetics* / Q000378:metabolism; D005169:Factor VIII / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006467:Hemophilia A / Q000235:genetics / Q000628:therapy*; D006801:Humans; D007422:Intestines / Q000166:cytology* / Q000821:virology; D008081:Liposomes; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D008081:Liposomes; D011994:Recombinant Proteins; D005169:Factor VIII; D005164:Factor IX