DOP80 Multi-omic sequencing reveals distinctive gene expression and DNA methylation alterations as potential predictors of primary sclerosing cholangitis development in treatment-naïve paediatric ulcerative colitis
Lawal, O.(1)*;Rodriguez-Sosa, A.(1);McDonnell, C.(1);Grant, L.(1);O'Brien, J.(2);Kel, A.(3);Dominik, A.(4);Stack, R.(5);Yochum, G.(6,7);Koltun, W.(6);Doherty, G.(5);Hussey, S.(8);
(1)Royal College of Surgeons in Ireland, School of Pharmacy and Biomolecular Sciences, Dublin, Ireland;(2)Children’s Health Ireland- Crumlin, Department of Pathology, Dublin, Ireland;(3)GeneXplain GmbH, GeneXplain GmbH, Wolfenbüttel, Germany;(4)Children’s Health Ireland- Crumlin, DOCHAS Study, Dublin, Ireland;(5)St. Vincent’s University Hospital and University College, Center for Colorectal Disease, Dublin, Ireland;(6)The Pennsylvania State University- College of Medicine- PA, Department of Surgery- Division of Colon and Rectal Surgery, Pennsylvania, United States;(7)The Pennsylvania State University- College of Medicine- PA, Department of Biochemistry & Molecular Biology, Pennsylvania, United States;(8)UCD and Royal College of Surgeons in Ireland, Department of Paediatrics, Dublin, Ireland;
Background
Primary sclerosing cholangitis (PSC) is a progressive choleostatic disease and up to 80% of patients also have ulcerative colitis (PSC-UC). This presents a clinical challenge owing to difficulty in diagnosis and increased risk for developing cancer. While several multifactorial processes including inflammation and microbial dysbiosis have been associated with PSC-UC pathogenesis, the precise molecular factors that regulate the phenotype of this disease subtype remain unknown.
Methods
Here, we applied methyl-capture sequencing and mRNA sequencing to colonic mucosal biopsies to identify transcriptomic and epigenetic alterations differentiating UC from PSC-UC. Samples were taken from 3 groups of treatment-naïve children at diagnosis who participated in the DOCHAS study (GEN-193/11) - UC (n=10), PSC-UC (n=10) and healthy controls(n=10).
Results
Differential gene expression between UC and PSC-UC identified disease-associated genes that were either up- or down-regulated in UC or PSC relative to controls. Furthermore, expression of these genes was intricately regulated by master transcriptional regulators (pro-caspases, IL7RA) and transcription factors (AR, p53, JUND, CEBPA). Importantly, gene expression comparison between UC vs PSC-UC revealed 4 up-regulated genes in PSC-UC (RAB31, TENM3, KLHL17 and COL7A1). Notably, RAB31 and TENM3 are also significantly up-regulated in gastrointestinal (GI) cancers. We also identified 4 down-regulated genes in PSC-UC (H3C15, SLC37A2, SLC14A2 and IL20RA).
Differential methylation analysis between healthy control biopsies vs PSC-UC and UC demonstrated >1000 differentially methylated regions (DMRs, 5Kb), with majority of these sites displaying hypermethylation. Interestingly, PSC-UC vs UC analysis identified 53 regions and 59 DMRs that are hyper- and hypomethylated respectively in PSC-UC, with a large proportion of these DMRs located in gene promoters and putative enhancer regions and notably impacted gene expression of differentially expressed genes identified from the mRNAseq analysis.Conclusion
Taken together, our study for the first time identifies distinct gene expression and DNA methylation alterations that differentiate UC from PSC-UC at diagnosis in treatment-naïve paediatric patients, some of which are associated with GI cancers. Their potential utility as predictive biomarkers of PSC development in UC or dysplasia risk in PSC-UC warrants further validation through larger patient cohorts.