Abstract
Background
The gastrointestinal (GI) microbiome is recognized for potential clinical relevance in inflammatory bowel disease (IBD). Data suggest that there is a disease-dependent loss of microbial diversity in IBD. Trimethylamine-N-oxide (TMAO) is generated by GI anaerobes through the digestion of dietary phosphatidylcholine and carnitine in a microbial-mammalian co-metabolic pathway. IBD-related changes in the gut microbiome may result in disease-specific changes in TMAO plasma concentrations.
Aim
To determine whether TMAO plasma levels in IBD are altered compared to controls and whether they correlate with disease presence or activity.
Methods
Liquid chromatography–tandem mass spectrometry was used to measure TMAO, choline, and carnitine plasma levels in 479 subjects (373 non-IBD controls, 106 IBD). Subjects were also genotyped for the flavin monooxygenase (FMO)3 variants, E158K and E308G.
Results
Plasma TMAO levels were 2.27 µM lower in the IBD population compared to the control population (p = 0.0001). Lower TMAO levels were similarly seen in active ulcerative colitis (UC) (1.56 µM) versus inactive disease (3.40 µM) (p = 0.002). No difference was seen in active Crohn’s disease (CD) versus inactive CD. No intergroup variation existed in plasma TMAO levels based on FMO3 genotype. Choline levels were higher in IBD, while carnitine levels were similar between the two groups, suggesting that lower TMAO levels in IBD were not due to dietary differences.
Conclusions
Decreased TMAO levels are seen in IBD compared to a non-IBD population. These data suggest that TMAO may have potential as a biomarker to support IBD diagnosis as well as to assess disease activity in UC.
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References
Molodecky NA, Soon IS, Rabi DM, et al. Increasing incidence and prevalence of the inflammatory bowel diseases with time, based on systematic review. Gastroenterology. 2012;142:9.
Ekbom A, Helmick C, Zack M, et al. The epidemiology of inflammatory bowel disease: a large, population-based study in Sweden. Gastroenterology. 1991;100:350–358.
Su C, Lichtenstein GR. Sleisenger and Fordtran’s gastrointestinal and liver diseases: pathophysiology/diagnosis/management, vol. 2. Philadelphia, PA: Saunders Elsevier; 2006.
Prideaux L, De Cruz P, Ng SC, et al. Serological antibodies in inflammatory bowel disease: a systematic review. Inflamm Bowel Dis. 2012;18:1340–1355.
Iskandar HN, Ciorba MA. Biomarkers in inflammatory bowel disease: current practices and recent advances. Trans Res J Lab Clin Med. 2012;159:313–325.
Lin JF, Chen JM, Zuo JH, et al. Meta-analysis: fecal calprotectin for assessment of inflammatory bowel disease activity. Inflamm Bowel Dis. 2014;20:1407–1415.
Vermeire S, Van Assche G, Rutgeerts P. C-reactive protein as a marker for inflammatory bowel disease. Inflamm Bowel Dis. 2004;10:661–665.
Sartor RB, Mazmanian SK. Intestinal microbes in inflammatory bowel diseases. Am J Gastroenterol Suppl. 2012;1:15–21.
Frank DN, St. Amand AL, Feldman RA, et al. Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases. Proc Natl Acad Sci USA. 2007;104:13780–13785.
Arumugam M, Raes J, Pelletier E, et al. Enterotypes of the human gut microbiome. Nature. 2011;473:174–180.
Hayashi H, Sakamoto M, Benno Y. Phylogenetic analysis of the human gut microbiota using 16s rDNA clone libraries and strictly anaerobic culture-based methods. Microbiol Immunol. 2002;46:535–548.
Lay C, Rigottier-Gois L, Holmstrom K, et al. Colonic microbiota signatures across five northern european countries. Appl Environ Microbiol. 2005;71:4153–4155.
Eckburg PB, Bik EM, Bernstein CN, et al. Diversity of the human intestinal microbial flora. Science (New York, NY). 2005;308:1635–1638.
Gill SR, Pop M, DeBoy RT, et al. Metagenomic analysis of the human distal gut microbiome. Science (New York, NY). 2006;312:1355–1359.
Wu GD, Chen J, Hoffmann C, et al. Linking long-term dietary patterns with gut microbial enterotypes. Science (New York, NY). 2011;334:105–108.
Yatsunenko T, Rey FE, Manary MJ, et al. Human gut microbiome viewed across age and geography. Nature. 2012;486:222–227.
Rajca S, Grondin V, Louis E, et al. Alterations in the intestinal microbiome (dysbiosis) as a predictor of relapse after infliximab withdrawal in Crohn’s disease. Inflamm Bowel Dis. 2014;20:978–986.
Ott SJ, Musfeldt M, Wenderoth DF, et al. Reduction in diversity of the colonic mucosa associated bacterial microflora in patients with active inflammatory bowel disease. Gut. 2004;53:685–693.
Takaishi H, Matsuki T, Nakazawa A, et al. Imbalance in intestinal microflora constitution could be involved in the pathogenesis of inflammatory bowel disease. Int J Med Microbiol (IJMM). 2008;298:463–472.
Schwiertz A, Jacobi M, Frick J-S, et al. Microbiota in pediatric inflammatory bowel disease. J Pediatr. 2010;157:240.e241–244.e241.
Willing BP, Dicksved J, Halfvarson J, et al. A pyrosequencing study in twins shows that gastrointestinal microbial profiles vary with inflammatory bowel disease phenotypes. Gastroenterology. 2010;139:1844.e1841–1854.e1841.
Koeth RA, Wang Z, Levison BS, et al. Intestinal microbiota metabolism of l-carnitine, a nutrient in red meat, promotes atherosclerosis. Nat Med. 2013;19:576–585.
Wang Z, Klipfell E, Bennett BJ, et al. Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature. 2011;472:57–63.
Tang WH, Wang Z, Levison BS, et al. Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk. N Engl J Med. 2013;368:1575–1584.
Ussher JR, Lopaschuk GD, Arduini A. Gut microbiota metabolism of l-carnitine and cardiovascular risk. Atherosclerosis. 2013;231:456–461.
Swann J, Richards S, Shen Q, et al. Culture-independent analysis of the human gut microbiota and their activities. New Jersey: Wiley; 2011.
Krueger SK, Williams DE. Mammalian flavin-containing monooxygenases: structure/function, genetic polymorphisms and role in drug metabolism. Pharmacol Ther. 2005;106:357–387.
Cashman JR, Akerman BR, Forrest SM, et al. Population-specific polymorphisms of the human FMO3 gene: significance for detoxication. Drug Metab Dispos Biol Fate Chem. 2000;28:169–173.
Treacy EP, Akerman BR, Chow LM, et al. Mutations of the flavin-containing monooxygenase gene (FMO3) cause trimethylaminuria, a defect in detoxication. Hum Mol Genet. 1998;7:839–845.
Zeisel SH, Wishnok JS, Blusztajn JK. Formation of methylamines from ingested choline and lecithin. J Pharmacol Exp Therap. 1983;225:320–324.
Craciun S, Balskus EP. Microbial conversion of choline to trimethylamine requires a glycyl radical enzyme. Proc Natl Acad Sci. 2012;109:21307–21312.
Katz R. Biomarkers and surrogate markers: an FDA perspective. NeuroRx. 2004;1:189–195.
Parikh NI, Vasan RS. Assessing the clinical utility of biomarkers in medicine. Biomark Med. 2007;1:419–436.
Wang Z, Levison B, Hazen JE, Donahue L, Li X, Hazen SL. Measurement of trimethylamine-n-oxide by stable isotope dilution liquid chromatography tandem mass spectrometry. Anal Biochem. 2014;455:35–40.
Stein J, Schröder O, Milovic V, et al. Mercaptopropionate inhibits butyrate uptake in isolated apical membrane vesicles of the rat distal colon. Gastroenterology. 1995;108:673–679.
Stein J, Zores M, Schröder O. Short-chain fatty acid (SCFA) uptake into caco-2 cells by a pH-dependent and carrier mediated transport mechanism. Eur J Nutr. 2000;39:121–125.
Schröder O, Opritz J, Stein J. Substrate and inhibitor specificity of butyrate uptake in apical membrane vesicles of the rat distal colon. Digestion. 2000;62:152–158.
Harvey RF, Bradshaw JM. A simple index of Crohn’s-disease activity. Lancet. 1980;1:514.
Walmsley RS, Ayres RC, Pounder RE, et al. A simple clinical colitis activity index. Gut. 1998;43:29–32.
R Core Team. R: A language and environment for statistical computing. Vienna, Austria: R Foundation for Statistical Computing; 2014. Retrieved from http://www.R-project.org.
Pinheiro J, Bates D, DebRoy S, Sarkar D, R Core Team. Nlme: linear and nonlinear mixed effects models. R package version 3.1-117. Vienna, Austria: R Foundation for Statistical Computing; 2014. Retrieved from http://CRAN.R-project.org/package=nlme.
von Elm E, Altman DG, Egger M, et al. Strobe initiative. The strengthening the reporting of observational studies in epidemiology (strobe)statement: guidelines for reporting observational studies. J Clin Epidemiol. 2008;61:344–349.
Patel N, Alkhouri N, Eng K, et al. Metabolomic analysis of breath volatile organic compounds reveals unique breathprints in children with inflammatory bowel disease: a pilot study. Aliment Pharmacol Ther. 2014;40:498–507.
Kostic AD, Xavier RJ, Gevers D. The microbiome in inflammatory bowel disease: current status and the future ahead. Gastroenterology. 2014;146:1489–1499.
Khor B, Gardet A, Xavier RJ. Genetics and pathogenesis of inflammatory bowel disease. Nature. 2011;474:307–317.
Morgan XC, Tickle TL, Sokol H, et al. Dysfunction of the intestinal microbiome in inflammatory bowel disease and treatment. Genome Biol. 2012;13:R79.
Guarner F, Malagelada J-R. Gut flora in health and disease. Lancet. 2003;361:512–519.
Zheng X, Xie G, Zhao A, et al. The footprints of gut microbial–mammalian co-metabolism. J Proteome Res. 2011;10:5512–5522.
Sepehri S, Kotlowski R, Bernstein CN, et al. Microbial diversity of inflamed and noninflamed gut biopsy tissues in inflammatory bowel disease. Inflamm Bowel Dis. 2007;13:675–683.
Kang S, Denman SE, Morrison M, et al. Dysbiosis of fecal microbiota in Crohn’s disease patients as revealed by a custom phylogenetic microarray. Inflamm Bowel Dis. 2010;16:2034–2042.
Manichanh C, Rigottier-Gois L, Bonnaud E, et al. Reduced diversity of faecal microbiota in Crohn’s disease revealed by a metagenomic approach. Gut. 2006;55:205–211.
Dawiskiba T, Deja S, Mulak A, et al. Serum and urine metabolomic fingerprinting in diagnostics of inflammatory bowel diseases. World J Gastroenterol (WJG). 2014;20:163–174.
Dhurandhar NV, Schoeller D, Brown AW, et al. Energy balance measurement: when something is not better than nothing. Int J Obes (Lond). 2014. doi:10.1038/ijo.2014.199.
David LA, Maurice CF, Carmody RN, et al. Diet rapidly and reproducibly alters the human gut microbiome. Nature. 2014;505:559–563.
Bennett BJ, de Aguiar Vallim TQ, Wang Z, et al. Trimethylamine-n-oxide, a metabolite associated with atherosclerosis, exhibits complex genetic and dietary regulation. Cell Metab. 2013;17:49–60.
Miller CA, Corbin KD, da Costa KA, et al. Effect of egg ingestion on trimethylamine-n-oxide production in humans: a randomized, controlled, dose-response study. Am J Clin Nutr. 2014;100:778–786.
Chen Y, Patel NA, Crombie A, et al. Bacterial flavin-containing monooxygenase is trimethylamine monooxygenase. Proc Natl Acad Sci USA. 2011;108:17791–17796.
Berrill JW, Green JT, Hood K, et al. Symptoms of irritable bowel syndrome in patients with inflammatory bowel disease: examining the role of sub-clinical inflammation and the impact on clinical assessment of disease activity. Aliment Pharmacol Ther. 2013;38:44–51.
Burgmann T, Clara I, Graff L, et al. The manitoba inflammatory bowel disease cohort study: prolonged symptoms before diagnosis—how much is irritable bowel syndrome? Clin Gastroenterol Hepatol. 2006;4:614–620.
Zschocke J, Kohlmueller D, Quak E, et al. Mild trimethylaminuria caused by common variants in FMO3 gene. Lancet. 1999;354:834–835.
Raimondi F, Santoro P, Barone MV, et al. Bile acids modulate tight junction structure and barrier function of Caco-2 monolayers via EGFR activation. Am J Physiol Gastrointest Liver Physiol. 2008;294:G906–G913.
Cipriani S, Mencarelli A, Chini MG, et al. The bile acid receptor GPBAR-1 (TGR5) modulates integrity of intestinal barrier and immune response to experimental colitis. PLoS ONE [Electronic Resource]. 2011;6:e25637.
Gadaleta RM, van Erpecum KJ, Oldenburg B, et al. Farnesoid X receptor activation inhibits inflammation and preserves the intestinal barrier in inflammatory bowel disease. Gut. 2011;60:10.
Attinkara R, Mwinyi J, Truninger K, et al. Association of genetic variation in the NR1H4 gene, encoding the nuclear bile acid receptor FXR, with inflammatory bowel disease. BMC Res Notes. 2012;5:12.
Nijmeijer RM, Gadaleta RM, van Mil SW, et al. Farnesoid X receptor (FXR) activation and FXR genetic variation in inflammatory bowel disease. PLoS ONE [Electronic Resource]. 2011;6:e23745.
Acknowledgments
We kindly thank all members of the Division of Gastroenterology, Department of Medicine at Western University and London Health Sciences Centre for their contribution to this study. This work was supported by the Wolfe Medical Research Chair in Pharmacogenomics (RBK) and the Canadian Institutes of Health Research (MOP-89753 to RBK) as well as the Drug Safety and Effectiveness Network (DSEN-PREVENT, FRN-117588 to RBK), and Cancer Care Ontario (CCO Research Chair Award (Tier-1) in Experimental Therapeutics to RBK).
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Wilson, A., Teft, W.A., Morse, B.L. et al. Trimethylamine-N-oxide: A Novel Biomarker for the Identification of Inflammatory Bowel Disease. Dig Dis Sci 60, 3620–3630 (2015). https://doi.org/10.1007/s10620-015-3797-3
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DOI: https://doi.org/10.1007/s10620-015-3797-3