title: Effective in vivo and ex vivo gene transfer to intestinal mucosa by VSV-G-pseudotyped lentiviral vectors.
abstract: BACKGROUND
  Gene transfer to the gastrointestinal (GI) mucosa is a therapeutic strategy which could prove particularly advantageous for treatment of various hereditary and acquired intestinal disorders, including inflammatory bowel disease (IBD), GI infections, and cancer.
  
  
  METHODS
  We evaluated vesicular stomatitis virus glycoprotein envelope (VSV-G)-pseudotyped lentiviral vectors (LV) for efficacy of gene transfer to both murine rectosigmoid colon in vivo and human colon explants ex vivo. LV encoding beta-galactosidase (LV-beta-Gal) or firefly-luciferase (LV-fLuc) reporter genes were administered by intrarectal instillation in mice, or applied topically for ex vivo transduction of human colorectal explant tissues from normal individuals. Macroscopic and histological evaluations were performed to assess any tissue damage or inflammation. Transduction efficiency and systemic biodistribution were evaluated by real-time quantitative PCR. LV-fLuc expression was evaluated by ex vivo bioluminescence imaging. LV-beta-Gal expression and identity of transduced cell types were examined by histochemical and immunofluorescence staining.
  
  
  RESULTS
  Imaging studies showed positive fLuc signals in murine distal colon; beta-Gal-positive cells were found in both murine and human intestinal tissue. In the murine model, beta-Gal-positive epithelial and lamina propria cells were found to express cytokeratin, CD45, and CD4. LV-transduced beta-Gal-positive cells were also seen in human colorectal explants, consisting mainly of CD45, CD4, and CD11c-positive cells confined to the LP.
  
  
  CONCLUSIONS
  We have demonstrated the feasibility of LV-mediated gene transfer into colonic mucosa. We also identified differential patterns of mucosal gene transfer dependent on whether murine or human tissue was used. Within the limitations of the study, the LV did not appear to induce mucosal damage and were not distributed beyond the distal colon.
authors.affiliation: Magee-Womens Research Institute, Division of Gastroenterology, Hepatology, and Nutrition, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Matsumoto
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kimura
authors.forename: Kazunori
authors.initials: K
authors.lastname: Haga
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kasahara
authors.forename: Peter
authors.initials: P
authors.lastname: Anton
authors.forename: Ian
authors.initials: I
authors.lastname: McGowan
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology / Q000378:metabolism; D003108:Colonic Diseases / Q000628:therapy; D004279:DNA, Viral / Q000032:analysis*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D016133:Polymerase Chain Reaction; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics*
chemical_list: D004279:DNA, Viral; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins
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