MicroRNA overexpression in slow transit constipation leads to reduced Na v 1.5 current and altered smooth muscle contractility

Amelia Mazzone, Peter R. Strege, Simon J. Gibbons, Constanza Alcaino, Vikram Joshi, Andrew J. Haak, Daniel J. Tschumperlin, Cheryl E. Bernard, Robert R. Cima, David W. Larson, Heidi K. Chua, Rondell P. Graham, Mona El Refaey, Peter J. Mohler, Yujiro Hayashi, Tamas Ordog, Stefan Calder, Peng Du, Gianrico Farrugia, Arthur Beyder

Research output: Contribution to journalArticle

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 Na V 1.5. 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 Na V 1.5. Within this set, a novel Na V 1.5 regulator, let-7f, resulted in decreased Na V 1.5 expression, current density and reduced motility of GI smooth muscle. These results suggest Na V 1.5 and miRNAs as novel diagnostic and potential therapeutic targets in STC.

Original languageEnglish (US)
Pages (from-to)868-876
Number of pages9
JournalGut
Volume69
Issue number5
DOIs
StatePublished - May 1 2020

Keywords

  • constipation
  • genetics
  • intestinal ion transport
  • intestinal motility
  • motility disorders

ASJC Scopus subject areas

  • Gastroenterology

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    Mazzone, A., Strege, P. R., Gibbons, S. J., Alcaino, C., Joshi, V., Haak, A. J., Tschumperlin, D. J., Bernard, C. E., Cima, R. R., Larson, D. W., Chua, H. K., Graham, R. P., El Refaey, M., Mohler, P. J., Hayashi, Y., Ordog, T., Calder, S., Du, P., Farrugia, G., & Beyder, A. (2020). MicroRNA overexpression in slow transit constipation leads to reduced Na v 1.5 current and altered smooth muscle contractility. Gut, 69(5), 868-876. https://doi.org/10.1136/gutjnl-2019-318747