1. Combined high-throughput library screening and next generation RNA sequencing uncover microRNAs controlling human cardiac fibroblast biology
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Angelika Pfanne, Lea Grote-Levi, Robert Geffers, Katharina Schimmel, Mira Jung, Stevan D. Stojanović, Ke Xiao, Saskia Mitzka, Cheng-Kai Huang, Christine S. Falk, Annette Just, Katharina Bock, Felix Kleemiß, Thomas Thum, Franziska Kenneweg, Jan Fiedler, Christian Bär, Martin H. Meyer, and HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
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0301 basic medicine ,Cardiac fibrosis ,MiR-20a-5p ,Computational biology ,030204 cardiovascular system & hematology ,Biology ,Autophagy-Related Protein 7 ,Deep sequencing ,miR-132 ,03 medical and health sciences ,0302 clinical medicine ,Fibrosis ,microRNA ,medicine ,Autophagy ,Humans ,Molecular Biology ,Gene Library ,Base Sequence ,Novel miRs ,Sequence Analysis, RNA ,Superoxide Dismutase ,Myocardium ,Forkhead Box Protein O3 ,High-Throughput Nucleotide Sequencing ,Fibroblasts ,medicine.disease ,CTGF ,MicroRNAs ,030104 developmental biology ,Gene Expression Regulation ,Inactivation, Metabolic ,Myocardial fibrosis ,Reactive Oxygen Species ,Cardiology and Cardiovascular Medicine ,Reactive oxygen species ,Signal Transduction - Abstract
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. European Research Council
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