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

Area of Science:

• Gene therapy
• Virology
• Dermatology

Background:

• Lentiviral vectors pseudotyped with viral envelope glycoproteins can modify tropism for gene delivery.
• Targeting human skin with gene therapy holds potential for treating genetic anomalies.

Purpose of the Study:

• To evaluate the efficiency of different pseudotyped human immunodeficiency virus (HIV)-based vectors for gene transfer in human skin xenografts.
• To identify the most effective viral glycoprotein for cutaneous gene delivery.

Main Methods:

• Pseudotyping HIV vectors with envelope glycoproteins from vesicular stomatitis virus (VSV-G), Zaire Ebola (EboZ) virus, murine leukemia virus (MuLV), lymphocytic choriomeningitis virus (LCMV), Rabies, or Mokola virus.
• Qualitative and quantitative assessment of transgene expression (LacZ reporter gene) in human skin xenografts.
• Double-labeling immunofluorescence for beta-galactosidase, involucrin, and integrin beta1.
• TaqMan PCR to determine genome copy number.

Main Results:

• High transgene expression was observed in dermal fibroblasts transduced with VSV-G, EboZ, or MuLV pseudotyped vectors.
• VSV-G, EboZ, and MuLV pseudotyped vectors effectively targeted quiescent epidermal stem cells, leading to transgene expression in differentiated progeny.
• VSV-G pseudotyped vector demonstrated the highest efficiency for cutaneous transduction, indicated by superior beta-galactosidase expression and genome copy number.

Conclusions:

• Pseudotyping lentiviral vectors with specific viral glycoproteins can achieve efficient gene transfer into human skin.
• VSV-G is the most potent viral glycoprotein for pseudotyping HIV-based vectors for effective gene delivery to epidermal stem cells in the skin.