Clinical Trial: Microbiota and Immune microEnvironment in Pouchitis

Study Status: Recruiting
Recruit Status: Recruiting
Study Type: Interventional

Official Title: Microbiota and Immune microEnvironment in Pouchitis: Randomized Controlled Trial Oral Administration of Lactobacillus Casei DG After Ileostomy Closure in Ileal Pouch Mucosa

Brief Summary:

Microbiota and innate immunity in pouchitis: predisposing factors and modulation of the inflammation with probiotics.

Around 20-25% of ulcerative colitis patients undergo restorative proctocolectomy with ileal pouch anal anastomosis. Pouchitis is an idiopathic inflammatory disease that may occur in ileal pouches. In our recent studies, we showed altered microbiota and innate immunity relationships in pouchitis. We plain to perform a double-blind, placebo-controlled trial probiotic therapy vs placebo starting at the time of ileostomy closure to evaluate the impact of microbiota that colonizes the pouch mucosa in the pathogenesis of pouchits, to determine how expression and activation status of the innate immunity system in different cell types and anatomical districts of pouch mucosa relate to microbiota population and follow-up the clinical outcome of anal pouches in light of microbiota-innate immune system interplay.

Our study will include three phases:

  1. analysis of the intestinal microbiota with High Throughput Sequencing Unit and anaerobes cultures
  2. characterization of innate immunity with TLR, NLR, nicotinic receptors and LPMC analysis
  3. assessment of microbiota and innate immune system in the ileal pouch before ileostomy closure, 2 months after ileostomy closure and after 1 year follow up.

Detailed Summary:

Study aims

The working hypothesis is that a disruption of gut microbiota homeostasis, either spontaneous or caused by the deregulation of the innate immunity machinery within the pouch mucosa, contributes to the onset of chronic/relapsing pouchitis. Taking into account that bacterial colonization of the pouch mucosa can be ideally studied since its generation, this can be considered as an ideal model to study the interplay between gut microbiota and innate immune system. Therefore, the goal of this Research Unit is to establish how dysbiosis and innate immunity machinery dysfunction are established and how this disfunction contributes to the onset and maintenance of pouchitis. To address these relevant pathophysiological issues in light of their therapeutic implications, we plan to perform a randomized double-blind study probiotics vs placebo in patients who will undergo colectomy and subsequent ileal pouch-anal anastomosis to enrolling the patients at the time of ileostomy colure to:

  1. evaluate composition and pathogenic properties of the microbiota that colonizes the pouch mucosa after ileostomy closure;
  2. determine how expression and activation status of the innate immunity system in different cell types and anatomical districts of pouch mucosa relate to ileostomy closure;
  3. follow-up the clinical outcome of anal pouches in light of microbiota-innate immune system interplay.

Study design In a two year period we calculated to enrol at least 32 patients who will be randomized at ileostomy closure in a group receiving placebo (16 patients) and in a group receiving lactobacillus casei GD oral supplementation for 8 weeks (16 patients). Mucosa samples will be harvested at ile
Sponsor: University of Padova

Current Primary Outcome: quantification of inflammatory cytokines in the ileal mucosa levels by Bio-Plex cytokine immunoassay [ Time Frame: 8 weeks ]

IL-1ß, IL-6, TNF-alpha


Original Primary Outcome: Same as current

Current Secondary Outcome:

  • quantify epithelial and leucocytes-derived anti-microbial defensins [ Time Frame: 8 weeks ]
    Def2, Def3; DEFA5; DEFA6 by quantitative RT-PCR
  • pouchitis episodes [ Time Frame: 12 months ]
    pouchitis episodes evaluated at PDAI >5
  • Relative abundance of bacterial phyla in faecal specimens [ Time Frame: 8 weeks ]
    Relative abundance of bacterial phyla in faecal specimens will be estimated by sequencing the PCR amplicons targeting 16S rRNA gene for the DNA samples extracted from each faecal specimen.
  • Systemic and local inflammatory status [ Time Frame: 12 months ]
    Systemic and local inflammatory state will be assessed at each experimental timeline by: erythrocyte sedimentation rate (ESR), white blood cell count (WBC), platelets blood count (PLT), CRP and fecal lactoferrin
  • Histological inflammatory severity [ Time Frame: 8 weeks ]
    Floren score
  • activation status of macrophages, dendritic cells, infiltrating lymphocytes [ Time Frame: 8 weeks ]
    assessment of activation status of macrophages, dendritic cells, infiltratinglymphocytes evaluating surface markers (i.e.) and intracellular cytokines pattern (i.e. TNF , IFN , IL4, IL10) by cytofluorimetric analysis.
  • analysis of TLRs network [ Time Frame: 8 weeks ]
    quantitative RT-PCR and immunohistochemistry


Original Secondary Outcome: Same as current

Information By: University of Padova

Dates:
Date Received: April 5, 2017
Date Started: October 31, 2016
Date Completion: April 30, 2019
Last Updated: April 27, 2017
Last Verified: April 2017