Identification of adeno-associated viral vectors suitable for intestinal gene delivery and modulation of experimental colitis.

Abstract

Effective gene transfer with sustained gene expression is an important adjunct to the study of intestinal inflammation and future therapy in inflammatory bowel disease. Recombinant adeno-associated virus (AAV) vectors are ideal for gene transfer and long-term transgene expression. The purpose of our study was to identify optimal AAV pseudotypes for transduction of the epithelium in the small intestine and colon, which could be used for studies in experimental colitis. The tropism and transduction efficiencies of AAV pseudotypes 1-10 were examined in murine small intestine and colon 8 wk after administration by real-time PCR and immunohistochemistry. The clinical and histopathological effects of IL-10-mediated intestinal transduction delivered by AAVrh10 were examined in the murine IL-10⁻/⁻ enterocolitis model. Serum IL-10 levels and IL-10 expression were followed by ELISA and real-time PCR, respectively. AAV pseudotypes 4, 7, 8, 9, and 10 demonstrated optimal intestinal transduction. Transgene expression was sustained 8 wk after administration and was frequently observed in enteroendocrine cells. Long-term IL-10 gene expression and serum IL-10 levels were observed following AAV transduction in an IL-10-/- model of enterocolitis. Animals treated with AAVrh10-IL-10 had lower disease activity index scores, higher colon weight-to-length ratios, and lower microscopic inflammation scores. This study identifies novel AAV pseudotypes with small intestine and colon tropism and sustained transgene expression capable of modulating mucosal inflammation in a murine model of enterocolitis.

• Authors: Steven Polyak, Annette Mach, Stacy Porvasnik, Lisa Dixon, Thomas Conlon, Kirsten E Erger, Andres Acosta, Amy J Wright, Martha Campbell-Thompson, Irene Zolotukhin, Clive Wasserfall, Cathryn Mah
• Journal: American journal of physiology. Gastrointestinal and liver physiology (Am J Physiol Gastrointest Liver Physiol) Vol. 302 Issue 3 Pg. G296-308 (Feb 01 2012) ISSN: 1522-1547 [Electronic] United States
• PMID: 22114116 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• IL10 protein, mouse
• Interleukin-10
• Green Fluorescent Proteins

Topics

• Adenoviridae (genetics)
• Animals
• Colon (metabolism, pathology)
• Enterocolitis (diagnosis, genetics, pathology, therapy)
• Gastric Mucosa (metabolism)
• Gene Dosage (genetics)
• Gene Transfer Techniques
• Genetic Therapy (methods)
• Genetic Vectors (administration & dosage, genetics)
• Genome, Viral (genetics)
• Green Fluorescent Proteins (genetics, metabolism)
• Inflammation (pathology, therapy)
• Interleukin-10 (administration & dosage, blood, genetics, therapeutic use)
• Intestinal Mucosa (metabolism)
• Intestine, Small (metabolism, pathology)
• Intestines (pathology)
• Liver (metabolism)
• Male
• Mice
• Mice, Inbred BALB C
• Mice, Inbred C57BL
• Mice, Knockout
• Transgenes (genetics)
• Viral Tropism (genetics)