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Original research
Immune response and barrier dysfunction-related proteomic signatures in preclinical phase of Crohn’s disease highlight earliest events of pathogenesis
  1. Haim Leibovitzh1,2,
  2. Sun-Ho Lee1,2,
  3. Juan Antonio Raygoza Garay1,2,
  4. Osvaldo Espin-Garcia3,
  5. Mingyue Xue1,
  6. Anna Neustaeter1,
  7. Ashleigh Goethel1,
  8. Hien Q Huynh4,
  9. Anne M Griffiths5,
  10. Dan Turner6,
  11. Karen L Madsen7,
  12. Paul Moayyedi8,
  13. A Hillary Steinhart1,2,
  14. Mark S Silverberg1,2,
  15. Colette Deslandres9,
  16. Alain Bitton10,
  17. David R Mack11,
  18. Kevan Jacobson12,
  19. Maria Cino13,
  20. Guy Aumais14,
  21. Charles N Bernstein15,
  22. Remo Panaccione16,
  23. Batia Weiss17,
  24. Jonas Halfvarson18,
  25. Wei Xu3,
  26. Williams Turpin1,2,
  27. Kenneth Croitoru1,2
  28. Crohn’s and Colitis Canada (CCC) Genetic, Environmental, Microbial (GEM) Project Research Consortium
    1. 1 Zane Cohen Centre for Digestive Diseases, Lunenfeld Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada
    2. 2 Division of Gastroenterology & Hepatology, University of Toronto Temerty Faculty of Medicine, Toronto, Ontario, Canada
    3. 3 Division of Biostatistics, University of Toronto Dalla Lana School of Public Health, Toronto, Ontario, Canada
    4. 4 Division of Gastroenterology and Nutrition, Department of Pediatrics, University of Alberta Faculty of Medicine & Dentistry, Edmonton, Alberta, Canada
    5. 5 IBD Center, Department of Paediatrics, Faculty of Medicine, University of Toronto, The Hospital for Sick Children, Toronto, Ontario, Canada
    6. 6 The Juliet Keidan Institute of Pediatric Gastroenterology and Nutrition, The Hebrew University of Jerusalem, Shaare Zedek Medical Center, Jerusalem, Israel
    7. 7 Department of Medicine, University of Alberta, Edmonton, Alberta, Canada
    8. 8 Department of Medicine, Farncombe Family Digestive Health Research Institute, McMaster University, Hamilton, Ontario, Canada
    9. 9 Division of Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, University of Montreal, Saint Justine Hospital, Montreal, Quebec, Canada
    10. 10 Division of Gastroenterology and Hepatology, McGill University Health Centre, Montreal, Quebec, Canada
    11. 11 Division of Gastroenterology, Hepatology & Nutrition, University of Ottawa, Children's Hospital of Eastern Ontario, Ottawa, Ontario, Canada
    12. 12 Canadian Gastro-Intestinal Epidemiology Consortium, British Columbia Children's Hospital Research Institute, University of British Columbia, British Columbia Children's Hospital, Vancouver, British Columbia, Canada
    13. 13 Department of Medicine, Division of Gastroenterology, University of Toronto, Toronto, Ontario, Canada
    14. 14 Department of Medicine, Montreal University, Hôpital Maisonneuve-Rosemont, Montreal, Quebec, Canada
    15. 15 University of Manitoba Inflammatory Bowel Disease Clinical and Research Centre and Department of Internal Medicine, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, Manitoba, Canada
    16. 16 Inflammatory Bowel Disease Clinic, Division of Gastroenterology and Hepatology of Gastroenterology, University of Calgary, Calgary, Alberta, Canada
    17. 17 Division of Pediatric Gastroenterology, Hepatology and Nutrition, Sheba Medical Center, Tel-Hashomer, Sackler Faculty of Medicine, Tel-Aviv University, The Edmond and Lily Safra Children's Hospital, Tel Aviv, Israel
    18. 18 School of Medical Sciences. Department of Gastroenterology, Örebro University, Orebro, Sweden
    1. Correspondence to Dr Kenneth Croitoru, Zane Cohen Centre for Digestive Diseases, Sinai Health System, Toronto, ON M5T 3L9, Canada; kcroitoru{at}


    Objective The measure of serum proteome in the preclinical state of Crohn’s disease (CD) may provide insight into biological pathways involved in CD pathogenesis. We aimed to assess associations of serum proteins with future CD onset and with other biomarkers predicting CD risk in a healthy at-risk cohort.

    Design In a nested case–control study within the Crohn’s and Colitis Canada Genetics Environment Microbial Project (CCC-GEM) cohort, which prospectively follows healthy first-degree relatives (FDRs), subjects who developed CD (n=71) were matched with four FDRs remaining healthy (n=284). Using samples at recruitment, serum protein profiles using the Olink Proximity Extension Assay platform was assessed for association with future development of CD and with other baseline biomarkers as follows: serum antimicrobial antibodies (AS: positive antibody sum) (Prometheus); faecal calprotectin (FCP); gut barrier function using the fractional excretion of lactulose-to-mannitol ratio (LMR) assay.

    Results We identified 25 of 446 serum proteins significantly associated with future development of CD. C-X-C motif chemokine 9 (CXCL9) had the highest OR with future risk of CD (OR=2.07 per SD, 95% CI 1.58 to 2.73, q=7.9e-5), whereas matrix extracellular phosphoglycoprotein had the lowest OR (OR 0.44, 95% CI 0.29 to 0.66, q=0.02). Notably, CXCL9 was the only analyte significantly associated with all other CD-risk biomarkers with consistent direction of effect (FCP: OR=2.21; LMR: OR=1.67; AS: OR=1.59) (q<0.05 for all).

    Conclusion We identified serum proteomic signatures associated with future CD development, reflecting potential early biological processes of immune and barrier dysfunction.


    Data availability statement

    No data are available.

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    • HL and S-HL are joint first authors.

    • WT and KC are joint senior authors.

    • Twitter @HaimLeiboIBD, @SunHoLee15, @MingyueXue, @hsteinhart, @guthealthmd, @RPanaccione, @KenCroitoru

    • Correction notice This article has been corrected since it published Online First. The second author's name has been corrected.

    • Collaborators The CCC-GEM Project Research Consortium is composed of: Maria Abreu, Paul Beck, Charles N. Bernstein, Kenneth Croitoru, Levinus A. Dieleman, Brian Feagan, Anne M. Griffiths, David S. Guttman, Kevan Jacobson, Gilaad Kaplan, Denis O. Krause*, Karen L. Madsen, John K. Marshall, Paul Moayyedi, Mark Ropeleski, Ernest Seidman*, Mark S. Silverberg, Scott Snapper, Andy Stadnyk, A. Hillary Steinhart, Michael Surette, Dan Turner, Thomas Walters, Bruce Vallance, Guy Aumais, Alain Bitton, Maria Cino, Jeff Critch, Lee Denson, Colette Deslandres, Wael El-Matary, Hans Herfarth, Peter Higgins, Hien Q. Huynh, Jeff Hyams, David R. Mack, Jerry McGrath, Anthony Otley, and Remo Panaccione. (* deceased). The CCC-GEM Project recruitment site directors include Maria Abreu, Guy Aumais, Robert Baldassano, Charles N. Bernstein, Maria Cino, Lee Denson, Colette Deslandres, Wael El-Matary, Levinus A. Dieleman, Anne M. Griffiths, Charlotte Hedin, Hans Herfarth, Peter Higgins, Seamus Hussey, Hien Q. Huynh, Kevan Jacobson, David Keljo, David Kevans, Charlie Lees, David R. Mack, John K. Marshall, Jerry McGrath, Sanjay Murthy, Anthony Otley, Remo Panaccione, Nimisha Parekh, Sophie Plamondon, Graham Radford-Smith, Mark Ropeleski, Joel Rosh, David Rubin, Michael Schultz, Ernest Seidman*, Corey Siegel, Scott Snapper, A. Hillary Steinhart, and Dan Turner. (* deceased)

    • Contributors Study guarantor: KC. Designed the study: HL, SHL, JARG, KM, AMG, PM, DT, MSS, AHS, KJ, DM, HQH, GA, MC, CD, RP, AB, CNB, JH, WT, KC and The CCC-GEM Project Research Consortium. Wrote the paper: HL, SHL, JARG, AN, AG, KM, AMG, PM, CNB, DT, JH, WT and KC. AnalySed data: HL, SHL, JARG, MX, KM, WT and KC. Subject recruitment: KM, AMG, PM, DT, AHS, MSS, KJ, DM, HQH, MC, GA, RP, AB, BW, CD, CNB, KC and The CCC-GEM Project Research Consortium Interpreted data: HL, SHL, JARG, WT and KC. Performed statistical analysis: HL, SHL, JARG, WT, WX and OE-G.

    • Funding This study was supported by grants from Crohn's and Colitis Canada Grant #CCC-GEMIII, Canadian Institutes of Health Research (CIHR) Grant #CMF108031 and The Leona M. and Harry B. Helmsley Charitable Trust. WT is a former recipient of a Postdoctoral Fellowship Research Award from the CIHR Fellowship/ Canadian Association of Gastroenterology (CAG)/ Ferring Pharmaceuticals. SHL is a recipient of the Imagine/CAG/CIHR Fellowship Award. HL is a former recipient of a fellowship award from the Israeli Association of Gastroenterology. SHL, JARG and WT are former recipients of a fellowship award from the Department of Medicine, Mount Sinai Hospital, Toronto. We thank the members of the CCC-GEM Global Project Office. KC is recipient of the Canada Research Chair in Inflammatory Bowel Diseases.

    • Competing interests None declared.

    • Patient and public involvement Patients and/or the public were not involved in the design, or conduct, or reporting, or dissemination plans of this research.

    • Provenance and peer review Not commissioned; externally peer reviewed.

    • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.