Metabolomics reveals elevated urinary excretion of collagen degradation and epithelial cell turnover products in irritable bowel syndrome patients
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Irritable bowel syndrome (IBS), the most commonly diagnosed functional gastrointestinal (GI) disorder in developed countries, is characterized by chronic abdominal pain, and altered bowel habits.
Accurate and timely diagnosis is challenging as it relies on symptoms and an evolving set of exclusion criteria to distinguish it from other related GI disorders reflecting a complex etiology that remains poorly understood. Herein, nontargeted metabolite profiling of repeat urine specimens collected from a cohort of IBS patients (n = 42) was compared to healthy controls (n = 20) to gain insights into the underlying pathophysiology.
An integrated data workflow for characterization of the urine metabolome with stringent quality control was developed to authenticate reliably measured (CV < 30%) and frequently detected (> 75%) metabolites using multisegment injection-capillary electrophoresis-mass spectrometry. Complementary statistical methods were then used to rank differentially excreted urinary metabolites after normalization to osmolality that were subsequently identified by high resolution tandem mass spectrometry and their electrophoretic migration behavior.
Our work revealed ten consistently elevated urinary metabolites in repeat samples collected from IBS patients at two different time points (q < 0.05 after age and Benjamini-Hochberg/FDR adjustment), which were associated with greater collagen degradation and intestinal mucosal turn-over processes likely due to low-grade inflammation. IBS-specific metabolites identified in urine included a series of hydroxylysine metabolites (O-glycosylgalactosyl-hydroxylysine, O-galactosyl-hydroxylysine, lysine), mannopyranosy-l-tryptophan, imidazole propionate, glutamine, serine, ornithine, dimethylglycine and dimethylguanosine. A major limitation in this retrospective case-control study was significant co-morbidity of IBS patients with other illnesses, including depression and prescribed medications as compared to healthy controls.
This work provides new mechanistic insights into the pathophysiology of IBS while also offering a convenient way to monitor patient disease progression and treatment responses to therapy based on a panel of urinary metabolites that avoids invasive blood sampling, colonoscopy and/or tissue biopsies.
KeywordsMetabolomics Irritable bowel syndrome Urine Biomarkers Mechanisms
P.B.M. acknowledges funding support from the Natural Sciences and Engineering Research Council of Canada, Genome Canada, and McMaster University. P.B. also acknowledges support of a Foundation Grant from the Canadian Institutes of Health Research. We also acknowledge kind support from David Arndt, Zachary Budinski and David S. Wishart at the University Alberta with their assistance in uploading MS/MS spectra for urinary metabolites onto HMDB (www.hmdb.ca) identified in this work.
MY prepared and analyzed all urine samples and conducted data pre-processing and final statistical analysis under the supervision of PBM. MY also wrote the initial draft of the manuscript with final edits performed by PBM. MIP-S and PB designed the study, recruited participants, collected patient urine samples and also provided feedback on the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
This study was approved by the Hamilton Integrated Research Ethics Board at McMaster University (REB Project #3992).
Informed consent was obtained from all individual participants included in this study.
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