Effective in vivo and ex vivogene transfer to intestinal mucosa by VSV-G-pseudotyped lentiviral vectors

BMC Gastroenterology

Research Article

Hiroshi Matsumoto¹  Kazunori Haga¹  Takahiro Kimura¹  Peter Anton¹  Noriyuki Kasahara²  Ian McGowan³ 

[1] Department of Medicine, Division of Digestive Diseases, University of California Los Angeles (UCLA) David Geffen School of Medicine, Los Angeles, CA, USA;Department of Medicine, Division of Digestive Diseases, University of California Los Angeles (UCLA) David Geffen School of Medicine, Los Angeles, CA, USA;Department of Molecular & Medical Pharmacology, University of California Los Angeles (UCLA) David Geffen School of Medicine, Los Angeles, CA, USA;Magee-Womens Research Institute, Division of Gastroenterology, Hepatology, and Nutrition, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA;

关键词: Lamina Propria;    Colon Tissue;    Murine Colon;    Human Explant;    Vesicular Stomatitis Virus Glycoprotein;   

DOI  :  10.1186/1471-230X-10-44

 received in 2009-06-30, accepted in 2010-05-11,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundGene 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.MethodsWe 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-β-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-β-Gal expression and identity of transduced cell types were examined by histochemical and immunofluorescence staining.ResultsImaging studies showed positive fLuc signals in murine distal colon; β-Gal-positive cells were found in both murine and human intestinal tissue. In the murine model, β-Gal-positive epithelial and lamina propria cells were found to express cytokeratin, CD45, and CD4. LV-transduced β-Gal-positive cells were also seen in human colorectal explants, consisting mainly of CD45, CD4, and CD11c-positive cells confined to the LP.ConclusionsWe 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.

【 授权许可】

Unknown   
© Matsumoto et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.