microRNA overexpression in slow transit constipation leads to reduced NaV1.5 current and altered smooth muscle contractility.

Abstract:

OBJECTIVE:This study was designed to evaluate the roles of microRNAs (miRNAs) in slow transit constipation (STC). DESIGN:All 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. RESULTS:The 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. CONCLUSIONS:A 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.

journal_name

Gut

journal_title

Gut

authors

Mazzone A,Strege PR,Gibbons SJ,Alcaino C,Joshi V,Haak AJ,Tschumperlin DJ,Bernard CE,Cima RR,Larson DW,Chua HK,Graham RP,El Refaey M,Mohler PJ,Hayashi Y,Ordog T,Calder S,Du P,Farrugia G,Beyder A

doi

10.1136/gutjnl-2019-318747

subject

Has Abstract

pub_date

2020-05-01 00:00:00

pages

868-876

issue

5

eissn

0017-5749

issn

1468-3288

pii

gutjnl-2019-318747

journal_volume

69

pub_type

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