Identification of adeno-associated viral vectors suitable for intestinal gene delivery and modulation of experimental colitis
| Autor: | Cathryn Mah, Clive Wasserfall, Stacy Porvasnik, Annette Mach, Irene Zolotukhin, Thomas J. Conlon, Lisa R. Dixon, Martha Campbell-Thompson, Amy J. Wright, Steven Polyak, Andres Acosta, Kirsten E. Erger |
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| Rok vydání: | 2012 |
| Předmět: |
Male
Colon Physiology Genetic Vectors Green Fluorescent Proteins Gene Dosage Genome Viral Gene delivery Biology Inflammatory bowel disease Adenoviridae law.invention Viral vector Mice law Physiology (medical) Intestine Small Gene expression medicine Animals Transgenes Intestinal Mucosa Colitis Inflammation Mice Knockout Mice Inbred BALB C Hepatology Enterocolitis Gene Transfer Techniques Gastroenterology Genetic Therapy medicine.disease Small intestine Interleukin-10 Intestines Mice Inbred C57BL Viral Tropism Interleukin 10 medicine.anatomical_structure Liver Gastric Mucosa Immunology Recombinant DNA |
| Zdroj: | American Journal of Physiology-Gastrointestinal and Liver Physiology. 302:G296-G308 |
| ISSN: | 1522-1547 0193-1857 |
| DOI: | 10.1152/ajpgi.00562.2010 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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