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Sustained Production of Erythropoietin in Mice by Human Keratinocytes Transduced With an Adenoassociated Virus Vector

Vincent Descamps, MD; Nadia Blumenfeld, MD; Yves Beuzard, MD; Michel Perricaudet, PhD

Arch Dermatol. 1996;132(10):1207-1211.

Abstract
Background and Design
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 β-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 β-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 longterm production of a therapeutic protein in mice. These results must now be extended to human primary keratinocytes.

Arch Dermatol. 1996;132:1207-1211

Author Affiliations
From the Department of Dermatology, Hôpital Bichat, Paris, France (Dr Descamps); Centre National de la Recherche Scientifique UA 1301, Villejuif, France (Drs Descamps and Perricaudet); and Institut National de la Santé et de la Recherche Médicale U91, Hôpital Henri-Mondor, Créteil, France (Drs Blumenfeld and Beuzard).

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