Abstract
Purpose
Crohn’s disease is a chronic gastrointestinal inflammatory disease with possible extraintestinal symptoms. There are predisposing genetic factors and even monogenic variants of the disorder. One of the possible genetic factors are variants of the DUOX2 gene. The protein product of the DUOX2 gene is a dual oxidase enzyme producing H2O2 in the bowel. Reduced H2O2 levels impact mucosal homeostasis and contribute to the development of inflammatory bowel disease. Thus far, only 19 patients with IBD with the DUOX2 variants have been described.
Methods
Here we present a case report of an adolescent female diagnosed at eleven years of age with IBD that was subsequently reclassified as Crohn’s disease. She was treated with immunosuppressants and biological therapy but experienced additional complications. Her peripheral blood lymphocyte DNA was studied using massive parallel sequencing. Detected variants were functionally studied.
Results
Whole exome sequencing found two novel DUOX2 gene variants: a de novo variant c.3646C>T; p.R1216W and a maternally inherited variant c.3391G>A; p.A1131T which were initially classified as variants of unknown significance. However, follow-up functional studies demonstrated that both DUOX2 variants led to impaired H2O2 generation, which led to their reclassification to the likely pathogenic class according to the ACMG.net. Therefore, we conclude that these variants are causative for the disease.
Conclusions
Identifying novel variants in patients with Crohn’s disease and their families is important for precision medicine approaches and understanding of the pathogenesis of likely “monogenic” rare forms of inflammatory bowel disease.
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Data availability
The datasets used and/or analyzed during this study are available from the corresponding author upon request.
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Acknowledgements
We thank Doctor Ulla G. Knaus and Professor Helmut Grasberger for providing plasmids used in the study. We want to thank Tom Secrest for English proofreading. We would also like to extend our gratitude to the patient’s family for their cooperation. This work was done under the auspices of ERN ITHACA.
Funding
Ministry of Health, Czech Republic [grant number 00064203]; Ministry of Youth Education and Sports, Czech Republic [grant number LM2018132] and the Charles University, Czech Republic [grant number GAUK 134121].
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Author Martin Schwarz and Matej Gazdarica contributed equally and share co-first authorship. Matej Gazdarica conceived, performed and interpreted functional studies and drafted their respective parts in the manuscript. Martin Schwarz, conceived, designed and drafted the rest of the manuscript, led genetic consultations with the patients and interpreted laboratory genetic testing results. Eva Froňková, Michael Svatoň and Anna Křepelová performed and interpreted laboratory genetic testing results. Jiří Bronský, Markéta Havlovicová, Milan Macek jr. substantively revised the manuscript. Jiří Bronský followed the patient in the pediatric gastroenterology clinic. All authors have agreed to be personally accountable for their contributions. All authors read and approved the final manuscript. The manuscript, including related data, figures and tables has not been previously published and that the manuscript is not under consideration elsewhere.
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The study was approved by the Internal Ethics Board of the Motol University Hospital and was carried out in line with the Declaration of Helsinki principles. Parents of the patient underwent genetic counseling and signed informed consent with research and publication of its results.
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Schwarz, M., Gazdarica, M., Froňková, E. et al. Functional studies associate novel DUOX2 gene variants detected in heterozygosity to Crohn’s disease. Mol Biol Rep 51, 399 (2024). https://doi.org/10.1007/s11033-024-09317-8
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DOI: https://doi.org/10.1007/s11033-024-09317-8