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Crypt-top and crypt-bottom colonic epithelial cell microRNA profiling reveals cell type-specific response in active and quiescent ulcerative colitis

Inciuraite, R.; Ramonaite, R.; Kupcinskas, J.; Dalgediene, I.; Kulokiene, U.; Kiudelis, V.; Varkalaite, G.; Zvirbliene, A.; Jonaitis, L. V.; Kiudelis, G.; Franke, A.; Juzenas, S.; Skieceviciene, J.

2022-09-26 gastroenterology
10.1101/2022.09.25.22280336 medRxiv
Show abstract

Background and AimsColonic epithelial barrier loss and dysfunction are one of the early events in ulcerative colitis (UC) and microRNAs (miRNAs) participate in its regulation. However, cell type-specific profile of miRNAs during inflammation in UC is still unknown. Thus, we aimed to perform miRNA profiling on colon tissue and epithelial cell levels in active and quiescent UC. MethodsSmall RNA-sequencing in colon tissue, crypt-bottom (CD44+), and crypt-top (CD66a+) colonic epithelial cell populations from two independent cohorts of UC patients (active and quiescent, n=74), and healthy individuals (n=50) was performed. Data analysis encompassed differential expression, weighted gene co-expression network (WGCNA), correlation, gene-set enrichment analyses (GSEA). ResultsIn colon tissue of active and quiescent UC, differentially expressed miRNAs were shown to be potentially involved in intestinal barrier integrity regulation. Consecutive analysis of crypt-bottom and crypt-top colonic epithelial cells revealed distinct miRNA expression patterns in response to UC-caused inflammation. GSEA indicated that differentially expressed epithelial miRNAs are commonly involved in inflammation- and intestinal barrier integrity-related processes (such as signalling of interleukin-4 and interleukin-13), while miRNA differences between cell populations might reflect their function, i.e., crypt-bottom cell miRNA target genes involved in regulation of cell differentiation. Finally, pro-inflammatory miRNA co-expression module correlating with endoscopic UC activity was defined not only in both epithelial cell populations, but also in the colon tissue. ConclusionsmiRNA expression patterns are colon epithelial cell population- and UC state-specific and correlate with endoscopic UC activity. Irrespective of the UC stage deregulated epithelial miRNAs are potentially involved in regulation of intestinal barrier integrity.

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