title: Functional Transcriptomics in Diverse Intestinal Epithelial Cell Types Reveals Robust MicroRNA Sensitivity in Intestinal Stem Cells to Microbial Status.
abstract: Gut microbiota play an important role in regulating the development of the host immune system, metabolic rate, and at times, disease pathogenesis. The factors and mechanisms that mediate interactions between microbiota and the intestinal epithelium are not fully understood. We provide novel evidence that microbiota may control intestinal epithelial stem cell (IESC) proliferation in part through microRNAs (miRNAs). We demonstrate that miRNA profiles differ dramatically across functionally distinct cell types of the mouse jejunal intestinal epithelium and that miRNAs respond to microbiota in a highly cell type-specific manner. Importantly, we also show that miRNAs in IESCs are more prominently regulated by microbiota compared with miRNAs in any other intestinal epithelial cell subtype. We identify miR-375 as one miRNA that is significantly suppressed by the presence of microbiota in IESCs. Using a novel method to knockdown gene and miRNA expressionex vivoenteroids, we demonstrate that we can knock down gene expression in Lgr5+IESCs. Furthermore, when we knock down miR-375 in IESCs, we observe significantly increased proliferative capacity. Understanding the mechanisms by which microbiota regulate miRNA expression in IESCs and other intestinal epithelial cell subtypes will elucidate a critical molecular network that controls intestinal homeostasis and, given the heightened interest in miRNA-based therapies, may offer novel therapeutic strategies in the treatment of gastrointestinal diseases associated with altered IESC function.
authors.affiliation: From the Curriculum in Genetics and Molecular Biology, bpeck@email.unc.edu.|Department of Genetics.
authors.forename: Bailey C E
authors.identifier: 0000-0002-4316-0966
authors.initials: BC
authors.lastname: Peck
authors.affiliation: Department of Nutrition, and.
authors.forename: Amanda T
authors.initials: AT
authors.lastname: Mah
authors.affiliation: Department of Genetics.
authors.forename: Wendy A
authors.initials: WA
authors.lastname: Pitman
authors.affiliation: Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599.
authors.forename: Shengli
authors.initials: S
authors.lastname: Ding
authors.affiliation: From the Curriculum in Genetics and Molecular Biology.|Department of Nutrition, and.|Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599.
authors.forename: P Kay
authors.initials: PK
authors.lastname: Lund
authors.affiliation: From the Curriculum in Genetics and Molecular Biology, Praveen_sethupathy@med.unc.edu.|Department of Genetics.
authors.forename: Praveen
authors.identifier: 0000-0002-7983-239X
authors.initials: P
authors.lastname: Sethupathy
mesh_terms: D000818:Animals; D005260:Female; D005856:Germ-Free Life; D049452:Green Fluorescent Proteins / Q000235:genetics; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism* / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics; D055758:SOX9 Transcription Factor / Q000235:genetics; D013234:Stem Cells / Q000166:cytology / Q000382:microbiology*; D059467:Transcriptome
chemical_list: D035683:MicroRNAs; D055758:SOX9 Transcription Factor; C527933:Sox9 protein, mouse; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: genomics; intestinal epithelium; microRNA (miRNA); microbiome; stem cells