MicroRNA-363 negatively regulates the left ventricular determining transcription factor HAND1 in human embryonic stem cell-derived cardiomyocytes

Posttranscriptional control of mRNA by microRNA (miRNA) has been implicated in the regulation of diverse biologic processes from directed differentiation of stem cells through organism development. We describe a unique pathway by which miRNA regulates the specialized differentiation of cardiomyocyte...

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Bibliographic Details
Published in:Stem cell research & therapy 2014-06, Vol.5 (3), p.75-75, Article 75
Main Authors: Wagh, Vilas, Pomorski, Alexander, Wilschut, Karlijn J, Piombo, Sebastian, Bernstein, Harold S
Format: Article
Language:English
Subjects:
Analysis
Basic Helix-Loop-Helix Transcription Factors - biosynthesis
Basic Helix-Loop-Helix Transcription Factors - genetics
Cell Differentiation - genetics
Cell Line
Embryonic Stem Cells - cytology
Gene Expression Regulation, Developmental - genetics
Genetic aspects
Heart Ventricles - cytology
Humans
Immunoblotting
In Situ Hybridization, Fluorescence
Induced Pluripotent Stem Cells - cytology
MicroRNAs - genetics
Myocytes, Cardiac - cytology
Real-Time Polymerase Chain Reaction
Stem cell research
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Summary:Posttranscriptional control of mRNA by microRNA (miRNA) has been implicated in the regulation of diverse biologic processes from directed differentiation of stem cells through organism development. We describe a unique pathway by which miRNA regulates the specialized differentiation of cardiomyocyte (CM) subtypes. We differentiated human embryonic stem cells (hESCs) to cardiac progenitor cells and functional CMs, and characterized the regulated expression of specific miRNAs that target transcriptional regulators of left/right ventricular-subtype specification. From >900 known human miRNAs in hESC-derived cardiac progenitor cells and functional CMs, a subset of differentially expressed cardiac miRNAs was identified, and in silico analysis predicted highly conserved binding sites in the 3'-untranslated regions (3'UTRs) of Hand-and-neural-crest-derivative-expressed (HAND) genes 1 and 2 that are involved in left and right ventricular development. We studied the temporal and spatial expression patterns of four miRNAs in differentiating hESCs, and found that expression of miRNA (miR)-363, miR-367, miR-181a, and miR-181c was specific for stage and site. Further analysis showed that miR-363 overexpression resulted in downregulation of HAND1 mRNA and protein levels. A dual luciferase reporter assay demonstrated functional interaction of miR-363 with the full-length 3'UTR of HAND1. Expression of anti-miR-363 in-vitro resulted in enrichment for HAND1-expressing CM subtype populations. We also showed that BMP4 treatment induced the expression of HAND2 with less effect on HAND1, whereas miR-363 overexpression selectively inhibited HAND1. These data show that miR-363 negatively regulates the expression of HAND1 and suggest that suppression of miR-363 could provide a novel strategy for generating functional left-ventricular CMs.
ISSN:1757-6512
1757-6512
DOI:10.1186/scrt464