Combined high-throughput library screening and next generation RNA sequencing uncover microRNAs controlling human cardiac fibroblast biology.
Autophagy
/ genetics
Autophagy-Related Protein 7
/ metabolism
Base Sequence
Fibroblasts
/ metabolism
Fibrosis
Forkhead Box Protein O3
/ genetics
Gene Expression Regulation
Gene Library
High-Throughput Nucleotide Sequencing
Humans
Inactivation, Metabolic
/ genetics
MicroRNAs
/ genetics
Myocardium
/ cytology
Reactive Oxygen Species
/ metabolism
Sequence Analysis, RNA
Signal Transduction
Superoxide Dismutase
/ metabolism
Autophagy
Cardiac fibrosis
MiR-20a-5p
Novel miRs
Reactive oxygen species
miR-132
Journal
Journal of molecular and cellular cardiology
ISSN: 1095-8584
Titre abrégé: J Mol Cell Cardiol
Pays: England
ID NLM: 0262322
Informations de publication
Date de publication:
01 2021
01 2021
Historique:
received:
27
08
2020
revised:
08
10
2020
accepted:
16
10
2020
pubmed:
1
11
2020
medline:
9
10
2021
entrez:
31
10
2020
Statut:
ppublish
Résumé
Myocardial fibrosis is a hallmark of the failing heart, contributing to the most common causes of deaths worldwide. Several microRNAs (miRNAs, miRs) controlling cardiac fibrosis were identified in recent years; however, a more global approach to identify miRNAs involved in fibrosis is missing. Functional miRNA mimic library screens were applied in human cardiac fibroblasts (HCFs) to identify annotated miRNAs inducing proliferation. In parallel, miRNA deep sequencing was performed after subjecting HCFs to proliferating and resting stimuli, additionally enabling discovery of novel miRNAs. In-depth in vitro analysis confirmed the pro-fibrotic nature of selected, highly conserved miRNAs miR-20a-5p and miR-132-3p. To determine downstream cellular pathways and their role in the fibrotic response, targets of the annotated miRNA candidates were modulated by synthetic siRNA. We here provide evidence that repression of autophagy and detoxification of reactive oxygen species by miR-20a-5p and miR-132-3p explain some of their pro-fibrotic nature on a mechanistic level. We here identified both miR-20a-5p and miR-132-3p as crucial regulators of fibrotic pathways in an in vitro model of human cardiac fibroblast biology.
Sections du résumé
BACKGROUND
Myocardial fibrosis is a hallmark of the failing heart, contributing to the most common causes of deaths worldwide. Several microRNAs (miRNAs, miRs) controlling cardiac fibrosis were identified in recent years; however, a more global approach to identify miRNAs involved in fibrosis is missing.
METHODS AND RESULTS
Functional miRNA mimic library screens were applied in human cardiac fibroblasts (HCFs) to identify annotated miRNAs inducing proliferation. In parallel, miRNA deep sequencing was performed after subjecting HCFs to proliferating and resting stimuli, additionally enabling discovery of novel miRNAs. In-depth in vitro analysis confirmed the pro-fibrotic nature of selected, highly conserved miRNAs miR-20a-5p and miR-132-3p. To determine downstream cellular pathways and their role in the fibrotic response, targets of the annotated miRNA candidates were modulated by synthetic siRNA. We here provide evidence that repression of autophagy and detoxification of reactive oxygen species by miR-20a-5p and miR-132-3p explain some of their pro-fibrotic nature on a mechanistic level.
CONCLUSION
We here identified both miR-20a-5p and miR-132-3p as crucial regulators of fibrotic pathways in an in vitro model of human cardiac fibroblast biology.
Identifiants
pubmed: 33127387
pii: S0022-2828(20)30303-5
doi: 10.1016/j.yjmcc.2020.10.008
pii:
doi:
Substances chimiques
FOXO3 protein, human
0
Forkhead Box Protein O3
0
MicroRNAs
0
Reactive Oxygen Species
0
Superoxide Dismutase
EC 1.15.1.1
ATG7 protein, human
EC 6.2.1.45
Autophagy-Related Protein 7
EC 6.2.1.45
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
91-100Informations de copyright
Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.