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

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

Research output: Contribution to journalArticle

9 Citations (Scopus)

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.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume302
Issue number3
DOIs
StatePublished - Feb 1 2012
Externally publishedYes

Fingerprint

Colitis
Interleukin-10
Dependovirus
Enterocolitis
Genes
Colon
Transgenes
Small Intestine
Tropism
Inflammation
Real-Time Polymerase Chain Reaction
Enteroendocrine Cells
Gene Expression
Serum
Inflammatory Bowel Diseases
Epithelium
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry
Weights and Measures

Keywords

  • Colon
  • GFP
  • IL-10
  • Small intestine

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology (medical)
  • Physiology
  • Hepatology

Cite this

Identification of adeno-associated viral vectors suitable for intestinal gene delivery and modulation of experimental colitis. / Polyak, Steven; Mach, Annette; Porvasnik, Stacy; Dixon, Lisa; Conlon, Thomas; Erger, Kirsten E.; Acosta, Andres; Wright, Amy J.; Campbell-Thompson, Martha; Zolotukhin, Irene; Wasserfall, Clive; Mah, Cathryn.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 302, No. 3, 01.02.2012.

Research output: Contribution to journalArticle

Polyak, S, Mach, A, Porvasnik, S, Dixon, L, Conlon, T, Erger, KE, Acosta, A, Wright, AJ, Campbell-Thompson, M, Zolotukhin, I, Wasserfall, C & Mah, C 2012, 'Identification of adeno-associated viral vectors suitable for intestinal gene delivery and modulation of experimental colitis', American Journal of Physiology - Gastrointestinal and Liver Physiology, vol. 302, no. 3. https://doi.org/10.1152/ajpgi.00562.2010
Polyak, Steven ; Mach, Annette ; Porvasnik, Stacy ; Dixon, Lisa ; Conlon, Thomas ; Erger, Kirsten E. ; Acosta, Andres ; Wright, Amy J. ; Campbell-Thompson, Martha ; Zolotukhin, Irene ; Wasserfall, Clive ; Mah, Cathryn. / Identification of adeno-associated viral vectors suitable for intestinal gene delivery and modulation of experimental colitis. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 2012 ; Vol. 302, No. 3.
@article{cc41634ec73b44c1a4296ab155fa08ed,
title = "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.",
keywords = "Colon, GFP, IL-10, Small intestine",
author = "Steven Polyak and Annette Mach and Stacy Porvasnik and Lisa Dixon and Thomas Conlon and Erger, {Kirsten E.} and Andres Acosta and Wright, {Amy J.} and Martha Campbell-Thompson and Irene Zolotukhin and Clive Wasserfall and Cathryn Mah",
year = "2012",
month = "2",
day = "1",
doi = "10.1152/ajpgi.00562.2010",
language = "English (US)",
volume = "302",
journal = "American Journal of Physiology - Renal Fluid and Electrolyte Physiology",
issn = "1931-857X",
publisher = "American Physiological Society",
number = "3",

}

TY - JOUR

T1 - Identification of adeno-associated viral vectors suitable for intestinal gene delivery and modulation of experimental colitis

AU - Polyak, Steven

AU - Mach, Annette

AU - Porvasnik, Stacy

AU - Dixon, Lisa

AU - Conlon, Thomas

AU - Erger, Kirsten E.

AU - Acosta, Andres

AU - Wright, Amy J.

AU - Campbell-Thompson, Martha

AU - Zolotukhin, Irene

AU - Wasserfall, Clive

AU - Mah, Cathryn

PY - 2012/2/1

Y1 - 2012/2/1

N2 - 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.

AB - 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.

KW - Colon

KW - GFP

KW - IL-10

KW - Small intestine

UR - http://www.scopus.com/inward/record.url?scp=84856206687&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84856206687&partnerID=8YFLogxK

U2 - 10.1152/ajpgi.00562.2010

DO - 10.1152/ajpgi.00562.2010

M3 - Article

C2 - 22114116

AN - SCOPUS:84856206687

VL - 302

JO - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

JF - American Journal of Physiology - Renal Fluid and Electrolyte Physiology

SN - 1931-857X

IS - 3

ER -