microRNA overexpression in slow transit constipation leads to reduced NaV1.5 current and altered smooth muscle contractility
| Autor: | Heidi K. Chua, Vikram Joshi, Gianrico Farrugia, Rondell P. Graham, Constanza Alcaino, Peter R. Strege, Mona El Refaey, Amelia Mazzone, Robert R. Cima, Tamas Ordog, Stefan Calder, Daniel J. Tschumperlin, David W. Larson, Peter J. Mohler, Simon J. Gibbons, Andrew J. Haak, Yujiro Hayashi, Arthur Beyder, Peng Du, Cheryl E. Bernard |
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| Rok vydání: | 2019 |
| Předmět: |
Adult
0301 basic medicine intestinal motility Colon Motility Biology Real-Time Polymerase Chain Reaction Article Sampling Studies Neurogastroenterology 03 medical and health sciences 0302 clinical medicine Downregulation and upregulation Reference Values microRNA Humans genetics RNA Messenger Aged Messenger RNA Biopsy Needle Gastroenterology Muscle Smooth constipation Smooth muscle contraction Middle Aged intestinal ion transport Immunohistochemistry Up-Regulation Cell biology MicroRNAs 030104 developmental biology Real-time polymerase chain reaction Gene Expression Regulation Cell culture Case-Control Studies motility disorders Female 030211 gastroenterology & hepatology Gastrointestinal Motility Microtubule-Associated Proteins Ex vivo Muscle Contraction |
| Zdroj: | Gut |
| ISSN: | 1468-3288 0017-5749 |
| Popis: | ObjectiveThis study was designed to evaluate the roles of microRNAs (miRNAs) in slow transit constipation (STC).DesignAll human tissue samples were from the muscularis externa of the colon. Expression of 372 miRNAs was examined in a discovery cohort of four patients with STC versus three age/sex-matched controls by a quantitative PCR array. Upregulated miRNAs were examined by quantitative reverse transcription qPCR (RT-qPCR) in a validation cohort of seven patients with STC and age/sex-matched controls. The effect of a highly differentially expressed miRNA on a custom human smooth muscle cell line was examined in vitro by RT-qPCR, electrophysiology, traction force microscopy, and ex vivo by lentiviral transduction in rat muscularis externa organotypic cultures.ResultsThe expression of 13 miRNAs was increased in STC samples. Of those miRNAs, four were predicted to target SCN5A, the gene that encodes the Na+ channel NaV1.5. The expression of SCN5A mRNA was decreased in STC samples. Let-7f significantly decreased Na+ current density in vitro in human smooth muscle cells. In rat muscularis externa organotypic cultures, overexpression of let-7f resulted in reduced frequency and amplitude of contraction.ConclusionsA small group of miRNAs is upregulated in STC, and many of these miRNAs target the SCN5A-encoded Na+ channel NaV1.5. Within this set, a novel NaV1.5 regulator, let-7f, resulted in decreased NaV1.5 expression, current density and reduced motility of GI smooth muscle. These results suggest NaV1.5 and miRNAs as novel diagnostic and potential therapeutic targets in STC. |
| Databáze: | OpenAIRE |
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