Distinct Microbiome in Pouchitis Compared to Healthy Pouches in Ulcerative Colitis and Familial Adenomatous Polyposis

Garrett C. Zella, Division of Pediatric Gastroenterology, Department of Pediatrics, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts
Elizabeth J. Hait, Children's Hospital Boston, Division of Gastroenterology and Nutrition, Boston, Massachusetts
Tiffany Glavan, University of California - Davis
Dirk Gevers, Microbial Sequencing Center, Broad Institute of MIT and Harvard, Cambridge, Massachusetts
Doyle V. Ward, Genome Sequencing and Analysis Program, Broad Institute of MIT and Harvard, Cambridge, Massachusetts
Christopher L. Kitts, California Polytechnic State University - San Luis Obispo
Joshua R. Korzenik, Gastrointestinal Unit, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts

Article comments

This is the pre-peer reviewed version of the following article: Distinct Microbiome in Pouchitis Compared to Healthy Pouches in Ulcerative Colitis and Familial Adenomatous Polyposis, Christopher L. Kitts, Inflammatory Bowel Disease, 17:5, Copyright © 2010 Crohn's & Colitis Foundation of America, Inc. and Wiley-Blackwell., which has been published in final form at http://dx.doi.org/10.1002/ibd.21460.

Abstract

Background: Pouchitis occurs in up to 50% of patients with ulcerative colitis (UC) undergoing ileal pouch anal anastomosis (IPAA). Pouchitis rarely occurs in patients with familial adenomatous polyposis (FAP) who undergo IPAA. Our aim was to compare mucosal and luminal flora in patients with UC-associated pouchitis (UCP), healthy UC pouches (HUC), and healthy FAP pouches (FAP).

Methods: Nineteen patients were enrolled in thi s cross-sectional study (nine UCP, tl1ree HUC, seven FAP). Patie nts with active pouchitis were identified using the Pouchitis Disease Activity Index (PDAI). Ileal pouch mucosal biopsies and fecal samples were analyzed with a 16S rDNA-based terminal restriction fr agment length polymorphism (TRFLP) approach. Pooled fecal DNA from four UCP and four FAP pouches were sequenced for further speciation.

Results: TRFLP data revealed statistically significant diffe rences in the mucosal and fecal microbiota between each group of patients. UCP samples exhibited significantly more TRFLP peaks matching Clostridium and Eubacterium genera compared to HUC and FAP pouches and fewer peaks matching Lactobacillus and Streptococcus genera compared to FAP. DNA Sanger sequencing of a subset of luminal samples revealed UCP having more identifiable sequences of Firmicutes (51.2% versus 2 1.2%) and Verrucomicrobia (20.2% versus 3.2%), and fewer Bacteroidetes (1 7.9% versus 60.5%) and Proteobacteria (9.8% versus 14.7%) compared to FAP.

Conclusions: The pouch microbial environment appears to be distinctly different in the settings of UC pouchitis, healthy UC, and FAP. These findings suggest that a dysbiosis may exist in pouchitis which may be central to understanding the disease.