title: Keratinocytes as a target for gene therapy. Sustained production of erythropoietin in mice by human keratinocytes transduced with an adenoassociated virus vector.
abstract: METHODS
  Keratinocytes are ideal targets for somatic gene therapy. Among the viral gene transfer systems, adenoassociated virus vectors have recently gained attention. We studied the feasibility of using adenoassociated virus-transduced human keratinocytes to provide a long-term, high-level production of a therapeutic factor after implantation in mice.
  
  
  RESULTS
  Transduction of HeLa cells by an adenoassociated virus vector was ascertained by transfer of the beta-galactosidase reporter gene, which was visualized by the blue staining of infected cells after fixation and coloring by X-Gal (the substrate of the reaction for beta-galactosidase activity). In a second step, 2 HeLa cell lines transduced with an AAV harboring the erythropoietin complementary DNA and producing high amounts of erythropoietin in vitro were isolated. After implantation in nude mice, a high-level and long-term increase in hematocrit (for the 1-month duration of the study) was found, which was correlated to the size of the induced tumor.
  
  
  CONCLUSIONS
  Adenoassociated virus-transduced HeLa keratinocytes provide high-level, stable, and long-term production of a therapeutic protein in mice. These results must now be extended to human primary keratinocytes.
authors.affiliation: Department of Dermatology, Hôpital Bichat, Paris, France.
authors.forename: V
authors.initials: V
authors.lastname: Descamps
authors.forename: N
authors.initials: N
authors.lastname: Blumenfeld
authors.forename: Y
authors.initials: Y
authors.lastname: Beuzard
authors.forename: M
authors.initials: M
authors.lastname: Perricaudet
mesh_terms: D000818:Animals; D000229:Dependovirus; D004921:Erythropoietin / Q000096:biosynthesis*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D014161:Transduction, Genetic
chemical_list: D004921:Erythropoietin