Gene transfer in human skin with different pseudotyped HIV-based vectors

• Published in: Gene therapy Vol. 14; no. 8; pp. 648 - 656
• Main Authors: HACHIYA, A, SRIWIRIYANONT, P, JAMES, W. M, KOBINGER, G. P, PATEL, A, SAITO, N, OHUCHI, A, KITAHARA, T, TAKEMA, Y, TSUBOI, R, BOISSY, R. E, VISSCHER, M. O
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01-04-2007
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Applied cell therapy and gene therapy; beta-Galactosidase - genetics; Biological and medical sciences; Biotechnology; Cell differentiation; Copy number; Dermis - metabolism; Ebolavirus - genetics; Escherichia coli - enzymology; Expression vectors; Fibroblasts; Fundamental and applied biological sciences. Psychology; Gene Expression; Gene therapy; Gene transfer; Genetic aspects; Genetic disorders; Genetic Therapy - methods; Genetic transformation; Genetic Vectors - administration & dosage; Genetic Vectors - genetics; Genomes; Glycoproteins; Health. Pharmaceutical industry; HIV; HIV - genetics; Human immunodeficiency virus; Humans; Immunofluorescence; Immunohistochemistry; Industrial applications and implications. Economical aspects; Leukemia Virus, Murine - genetics; Lymphocytic choriomeningitis virus; Lymphocytic choriomeningitis virus - genetics; Medical sciences; Mice; Mice, Nude; Microscopy, Confocal; Mokola virus; Murine leukemia virus; Physiological aspects; Rabies; Rabies virus - genetics; Reporter gene; Retroviridae - genetics; Skin; Skin Diseases - metabolism; Skin Diseases - therapy; Stem cell transplantation; Stem cells; Stomatitis; Transduction, Genetic - methods; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Transgenes; Transplantation, Heterologous; Tropism; Vesicular stomatitis Indiana virus - genetics; Vesicular stomatitis virus; Viral Envelope Proteins - genetics; Viruses; Xenografts; β-Galactosidase; Japan; Human; stem cells; Stem cell; Glycoprotein; Retroviridae; Lentivirus; Genetic transfer; Virus; Vesicular stomatitis virus; Mononegavirales; Vesiculovirus; vesicular stomatitis virus glycoprotein; Rhabdoviridae; Skin; Gene therapy; Vector; pseudotype
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3302915

Pseudotyping lentiviral vector with other viral surface proteins could be applied for treating genetic anomalies in human skin. In this study, the modification of HIV vector tropism by pseudotyping with the envelope glycoprotein from vesicular stomatitis virus (VSV), the Zaire Ebola (EboZ) virus, murine leukemia virus (MuLV), lymphocytic choriomeningitis virus (LCMV), Rabies or the rabies-related Mokola virus encoding LacZ as a reporter gene was evaluated qualitatively and quantitatively in human skin xenografts. High transgene expression was detected in dermal fibroblasts transduced with VSV-G-, EboZ- or MuLV-pseudotyped HIV vector with tissue irregularities in the dermal compartments following repeated injections of EboZ- or LCMV-pseudotyped vectors. Four weeks after transduction, double-labeling immunofluorescence of beta-galactosidase and involucrin or integrin beta1 demonstrated that VSV-G-, EboZ- or MuLV-pseudotyped HIV vector effectively targeted quiescent epidermal stem cells which underwent terminal differentiation resulting in transgene expression in their progenies. Among the six different pseudotyped HIV-based vectors evaluated, VSV-G-pseudotyped vector was found to be the most efficient viral glycoprotein for cutaneous transduction as demonstrated by the highest level of beta-galactosidase expression and genome copy number evaluated by TaqMan PCR.