Cellular and molecular mechanisms of inflammation in the GI tract
Research type
Research Study
Full title
Cellular and Molecular Mechanisms of Inflammation in the Gastrointestinal Tract
IRAS ID
250261
Contact name
Mark Pritchard
Contact email
Sponsor organisation
University of Liverpool
Duration of Study in the UK
2 years, 11 months, 11 days
Research summary
Research Summary:
Inflammatory Bowel Disease (IBD) is a chronic inflammatory condition of the small and large intestines. Currently, the exact cause of IBD is unclear but evidence point towards a complex interplay between host genetic susceptibility, immune system, and bacterial community.
This study aims to create a library of stem cell cell-lines over the course of 3 years from tissue samples collected from NHS patients at the Royal Liverpool University Hospital already schedule for endoscopy (camera tests) or an operation for the removal their large bowel. This will then allow comparison between patients with IBD and non-IBD conditions such as anaemia. Samples collected from these patients will be anonymised using a study ID and a separate database will be generated with basic patient demographics e.g. gender, age, and diagnose.Some of the tissue collected will be used to compare the gene and protein expressions of key components in inflammatory signalling pathways such as the alternative NF-κB signalling pathway. The remaining tissue will be used to generate intestinal stem cell cell-lines. 3-dimentional culture of these cell-lines will enable the generation of intestinal organoids (mini-gut system), which compose of a signal layer of cells arranged in the same configuration as gut epithelium (innermost layer). These organoids will serve as a laboratory model of the human gut to help researchers understand how inflammation occurs, how best to suppress or even prevent it. Furthermore, these organoids will be invaluable in assessing drug response and toxicity for potential therapeutic targets.
With consent, blood samples will be taken from these patients to measure markers of inflammation and disease severity. Furthermore, we aim to use a proportion of the blood sample to generate a library of induced pluripotent stem (IPS) cells to allow future studies to generate other organ-specific stem cells e.g. liver stem cells.
Summary of Results:
Fifteen patients donated colonic biopsy samples and after making some alterations to the tissue culture technique, colonic organoid cultures were successfully established. However we only managed to culture 2 samples successfully to permit treatment with the pro-inflammatory cytokine Tumour Necrosis Factor (TNF) in the culture dish. This treatment did not alter the size and shape of human colonic organoids as we had previously seen in mouse colonic organoids, but it did alter the expression of some inflammatory genes. Patient recruitment was halted as a result of the COVID pandemic and never restarted. The results of our studies are very preliminary and therefore no major conclusions can be drawn from this work.
REC name
North West - Liverpool Central Research Ethics Committee
REC reference
19/NW/0038
Date of REC Opinion
11 Mar 2019
REC opinion
Favourable Opinion