Combined high-throughput library screening and next generation RNA sequencing uncover microRNAs controlling human cardiac fibroblast biology

• Published in: Journal of molecular and cellular cardiology Vol. 150; pp. 91 - 100
• Main Authors: Schimmel, Katharina, Stojanović, Stevan D., Huang, Cheng-Kai, Jung, Mira, Meyer, Martin H., Xiao, Ke, Grote-Levi, Lea, Bär, Christian, Pfanne, Angelika, Mitzka, Saskia, Just, Annette, Geffers, Robert, Bock, Katharina, Kenneweg, Franziska, Kleemiß, Felix, Falk, Christine S., Fiedler, Jan, Thum, Thomas
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.01.2021
• Subjects: Autophagy; Autophagy - genetics; Autophagy-Related Protein 7 - metabolism; Base Sequence; Cardiac fibrosis; Fibroblasts - metabolism; Fibrosis; Forkhead Box Protein O3 - genetics; Forkhead Box Protein O3 - metabolism; Gene Expression Regulation; Gene Library; High-Throughput Nucleotide Sequencing; Humans; Inactivation, Metabolic - genetics; MicroRNAs - genetics; MicroRNAs - metabolism; miR-132; MiR-20a-5p; Myocardium - cytology; Novel miRs; Reactive oxygen species; Reactive Oxygen Species - metabolism; Sequence Analysis, RNA; Signal Transduction; Superoxide Dismutase - metabolism; MiR/MiRNA; Reactive oxygen species; RNA; HCF; pre-miR; SOD2; IL-8; Autophagy; CTGF; Novel miRs; COL1A1; pri-miR; Cardiac fibrosis; miR-132; ROS; AGO2; MiR-20a-5p; FOXO3a; BLASTN; ATG7; HF; LNA
• ISSN: 0022-2828; 1095-8584
• DOI: 10.1016/j.yjmcc.2020.10.008

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. [Display omitted] •A high throughput miRNA library screen can identify anti-fibrotic molecules•Autophagy and detoxification of reactive oxygen species play a major role in the miRNA-mediated fibrosis process•We identified potential druggable small noncoding RNA targets for the treatment of cardiac fibrosis