Molecular Gastroenterology Research Group

Overview

The Molecular Gastroenterology Research Group, part of the Academic Unit of Surgical Oncology, focuses on alterations in gut (primarily our focus is on the large bowel) causal or consequent to prevalent bowel conditions (colorectal cancer, colorectal polyps/adenoma, Inflammatory Bowel Disease (IBD), Irritable Bowel Syndrome (IBS). A particular focus of our work is the response of the musosa to the short-chain fatty acid butyrate, a compound which has been implicated in the chemopreventive properties of fibre, and which is in clinical trials, as an HDAC inhibitor, for the treatment of cancers. Proteins which are acetylated and may mediate apoptotic or cell cycle cellular responses are of particular interest in the modelling of HDAC inhibitor function and our work has addressed the biology of acetylation of Keratins 8 and 18 and of transcription factor Sp1. We have an interest in diet-gut interations and how these may be modified by disease, and the effect that disease may have on the gut response to the metabolic and nutritional environment, which we believe may be important in stratified medical approaches to the management and prevention of disease. The management of appetite and effects of disease on feeding behaviour forms the basis of a series of related studies.
We use an integrative approach, combining high-throughput Omics approaches (mainly proteomics and cellomics) and using a combination of in vitro models, cross-sectional studies and intervention trials in humans. This leads to a discovery cycle of modelling and testing through shuttling between in vitro and in vivo systems.

Biology of keratins in the colorectal mucosa

Biology of keratins


Intermediate filaments (IF) are characterizing cytoskeletal features of epithelia, with keratins 8, 18 and 19 expressed in the simple epithelia of the lower gut. We have shown that keratins 8 and 18 are acetyl proteins and that their acetylation is responsive to butyrate treatment, which triggers depolymerisation of the IF. We have developed a high-throughput assay to screen for drugs with IF depolymerising potential. Related in vivo work on field cancerization has shown that there are alterations in keratin 8 and 19 in subjects with colorectal neoplasia, and furthermore that there are alterations in the keratin response to butyrate in vivo. Published data has shown that genetic markers associated with IBD map to the keratin locus and our current programme of work addresses the alterations (expression, acetylation, proteolytic cleavage) resulting from duration, inflammatory status and cancer-predisposition in ulcerative colitis and in colorectal adenomagenesis.
Researchers: Ms Joanna Chowdry, Dr Debabrata Majumdar, Ms Ria Rosser
Collaborators: Imagen Biotech, Dr Alan Lobo, Mr Keith Chapple
Publications:
• Kahn AQ, Bury JP, Brown SR, Riley SA & Corfe BM (2011) Effect of butyrate and cancer progression on keratin 8 expression in the colon. BMC Gastroenterology 11: 2
• Drake PJM, Griffiths GJ, Benson RP & Corfe BM (2009) Application of high-content analysis to the study of post-translational modifications of the cytoskeleton. Journal of Proteome Research 8(1):28-34.
• Leech SH, Evans CA, Shaw L, Wong CH, Connolly J, Griffiths JR, Whetton AD & Corfe BM (2008) Proteomic analyses of intermediate filaments reveals cytokeratin 8 is highly acetylated – implications for colorectal epithelial homeostasis. Proteomics 8: 279-288
• Khan AQ, Shaw L, Drake PJM, Brown SR & Corfe BM. Application of high content biology demonstrates differential responses of keratin acetylation to short chain fatty acids. Journal of Integrated Omics –in press

Field changes in the pre-malignant colon

Field changes


There is a substantive body of literature to show that risk of recurrence of an adenoma is greater than risk of first incidence, however there remains a very marginal evidence-base of mechanistic data as to why this is the case. There is a great deal of modelling work addressing how mutations in stem cells in a crypt may come to expand and populate other crypts, forming morphologically normal pre-malignant fields which collide to license formation of polyclonal polyps, however our work suggests that the presence of a polyp or cancer may in turn drive effects in the mucosa which alter the response to the microenvironment. Our ongoing work aims to catalogue changes in the morphologically normal mucosa in subjects with lesions and uses proteomic approaches to identify field changes and their possible effectors. We are initiating a prospective study to assess whether such changes are a result of epigenetic modifications in the colon.
Researchers: Ria Rosser
Collaborators: Dr Stuart Riley, Dr Jonathan Bury, Mr Keith Chapple, Dr Carolyn Staton, Dr Caroline Evans
Publications:
• Yu CW, Bury JP, Tiernan JP, Waby JS, Staton CA & Corfe BM. (2011) Interaction between butyrate and field effects in the regulation of enteroendocrine cell number and neuropilin expression in the human colon epithelium. Molecular Cancer
• Kahn AQ, Bury JP, Brown SR, Riley SA & Corfe BM (2011) Effect of butyrate and cancer progression on keratin 8 expression in the colon. BMC Gastroenterology 11: 2

Systems biology of the colon lumen and host response to short-chain fatty acids

(i) Systems biology of the colon lumen
Our data and those of others suggest that the large gut microbiome may play at a more important role than diet in determination of the levels of short-chain fatty acids, which in turn may be key determinants of cell turnover in the mucosa. This project aims to build a model of the lumen accounting for dietary intakes, using high-throughput sequencing to catalogue bacterial strains, directly analysing SCFA levels and assessing the metabolome. Taken together we aim to reconcile the datastreams better to understand the production of butyrate and covarying molecules, in order to improve interventions aimed at benefitting health. The work stems from ongoing studies showing non-relationship between fibre intake and SCFA level, and beneficial effects of probiotics in IBS.
(ii) Systems biology of the host response to short-chain fatty acids
A related project is modelling the host cell management of butyrate from uptake through metabolism and HDAC inhibition. This project in particular aims to question paradigms and identify major gaps in understanding of butyrate handling in the eukaryotic cell. This includes the response of the cell through reprogramming gene expression by modification of transcription factor activity.
Researchers: Ms Amanda McLoone, Ms Yvonne Lennighan, Dr Jo Kilner, Bernard Corfe
Collaborators: Dr Julian Marchesi, Obsidian Research Ltd,
Publications:
• Kilner J, Corfe BM & Wilkinson S. (2011) Modelling the Microtubule: Towards a Better Understanding of Short-Chain Fatty Acid Molecular Pharmacology. Molecular BioSystems 7:975-983
• Waby JS, Chirakkal H, Yu CW, Griffiths GJ, Benson RSP, Bingle CD, Corfe BM. (2010) Sp1 acetylation results in loss of DNA binding at promoters associated with cell cycle arrest and cell death in a colon cell line. Molecular Cancer 9: 275
• Yu CW, Waby JS, Chirakkal H, Staton CA & Corfe BM. (2010) Butyrate suppresses expression of neuropilin I in colorectal cell lines through inhibition of Sp1 transactivation. Molecular Cancer 9:276
• Chirakkal H, Leech SH, Brookes KE, Prais AL, Waby JS, Corfe BM. (2006) Upregulation of BAK by butyrate in the colon is associated with increased Sp3 binding. Oncogene 25: 7192-200.
• Barker ME, Short RJ, Russell J & Corfe BM "A meta-analysis shows no dose-response relationship between fibre intake and faecal SCFA level. submitted to Am J Clin Nutr
• Williams E, Stimpson J, Wang D, Plummer S, Garaiova I, Barker M, Corfe BM (2009) Clinical trial: a multistrain probiotic preparation significantly reduces symptoms of irritable bowel syndrome in a double-blind placebo-controlled study. Alimentary Pharmacology and Therapy PMID: 18785988

Impact of diet on gut health, impact of gut health on diet

The gut is the first point of contact for ingesta and interacts and is affected by diet. Our work has suggested that enteroendocrine populations may be altered with disease state, and that individuals with high % body fat may have altered physiological responses to nutrient interventions. This body of work aims to investigate the bidirectionality of the relationship between diet and gut health.
Researchers: Mr Jon Wild, Mrs Charlotte Harden, Ms Amanda McLoone, Ms Yvonne Lennighan
Collaborators: Dr Margo Barker
Publications:
• Williams EA, Nai XL, Corfe BM. (2011) Dietary intakes in subjects with irritable bowel syndrome. BMC Gastroenterology 11:9
• Gunarwardene AR, Corfe BM & Staton CA (2011) Classification and Functions of Enteroendocrine Cells of the Lower GI Tract. International Journal of Experimental Pathology DOI: 10.1111/j.1365-2613.2011.00767.x
• Nakano E, Mushtaq S, Heath PR, Lee ES, Bury JP, Riley SA, Powers HJ & Corfe BM (2011) Riboflavin depletion impairs cell proliferation in intestinal cells – identification of mechanisms and consequences Digestive Diseases and Sciences 56: 1007-1019
• Harden C, Maya Jiminez T, Jones A, Barker ME, Hepburn NH, Garaiova I, Plummer S & Corfe BM "Use of an in vitro screen to identify long chain fatty acids triggering optimal CCK release and impact of Long chain fatty acids on 24-hr energy intake in healthy males" Br J Nutrition revision submitted
• Harden C, Richardson JC, Dettmar PW, Corfe BM, Paxman JR. An ionic gelling alginate formulation attenuates postprandial glycaemia in male subjects" J Funct Foods, in revision
• Harden C, Corfe BM, Richardson JC, Dettmar PW & Paxman JR. (2009) BMI and age affect Three Factor Eating Questionnaire scores in male subjects. Nutrition Res. 29:379-82
• Paxman JR, Richardson JC, Dettmar PW, Corfe BM. (2008) Alginate reduces the increased uptake of cholesterol and glucose in overweight male subjects: a pilot study. Nutr Res. 28:501-5.