title: Deciphering the microRNA signature of pathological cardiac hypertrophy by engineered heart tissue- and sequencing-technology.
abstract: Pathological cardiac hypertrophy and fibrosis are modulated by a set of microRNAs, most of which have been detected in biologically complex animal models of hypertrophy by arrays with moderate sensitivity and disregard of passenger strand (previously "star") microRNAs. Here, we aimed at precisely analyzing the microRNA signature of cardiac hypertrophy and fibrosis by RNA sequencing in a standardized in vitro hypertrophy model based on engineered heart tissue (EHT). Spontaneously beating, force-generating fibrin EHTs from neonatal rat heart cells were subjected to afterload enhancement for 7days (AE-EHT), and EHTs without intervention served as controls. AE resulted in reduced contractile force and relaxation velocity, fibrotic changes and reactivation of the fetal gene program. Small RNAs were extracted from control and AE-EHTs and sequencing yielded almost 750 different mature microRNAs, many of which have never been described before in rats. The detection of both arms of the precursor stem-loop (pre-miRNA), namely -3p and -5p miRs, was frequent. 22 abundantly sequenced microRNAs were >1.3× upregulated and 15 abundantly sequenced microRNAs downregulated to <0.77×. Among the upregulated microRNAs were 3 pairs of guide and passenger strand microRNAs (miR-21-5p/-3p, miR-322-5p/-3p, miR-210-3p/-5p) and one single passenger strand microRNA (miR-140-3p). Among downregulated microRNAs were 3 pairs (miR-133a-3p/-5p, miR-30e-5p/3p, miR-30c-5p/-3p). Preincubating EHTs with anti-miR-21-5p markedly attenuated the AE-induced contractile failure, cardiomyocyte hypertrophy and fibrotic response, recapitulating prior results in whole animals. Taken together, AE-induced pathological hypertrophy in EHTs is associated with 37 differentially regulated microRNAs, including many passenger strands. Antagonizing miR-21-5p ameliorates dysfunction in this model.
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Germany.
authors: Marc N
authors: MN
authors: Hirt
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Germany.
authors: Tessa
authors: T
authors: Werner
authors: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors: Daniela
authors: D
authors: Indenbirken
authors: Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany; Bioinformatics Service Facility, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Malik
authors: M
authors: Alawi
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Paul
authors: P
authors: Demin
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Germany.
authors: Ann-Cathrin
authors: AC
authors: Kunze
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Germany.
authors: Justus
authors: J
authors: Stenzig
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Germany.
authors: Jutta
authors: J
authors: Starbatty
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Germany.
authors: Arne
authors: A
authors: Hansen
authors: Institute of Molecular and Translational Therapeutic Strategies, Hannover Medical School, Hannover, Germany.
authors: Jan
authors: J
authors: Fiedler
authors: Institute of Molecular and Translational Therapeutic Strategies, Hannover Medical School, Hannover, Germany.
authors: Thomas
authors: T
authors: Thum
authors: Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany; DZHK (German Centre for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck, Germany. Electronic address: t.eschenhagen@uke.de.
authors: Thomas
authors: T
authors: Eschenhagen
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D006332:Cardiomegaly / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D005355:Fibrosis; D018507:Gene Expression Regulation, Developmental; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008955:Models, Cardiovascular; D009200:Myocardial Contraction / Q000235:genetics*; D020320:Oligoribonucleotides, Antisense / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011917:Rats, Inbred Lew; D017208:Rats, Wistar; D017423:Sequence Analysis, RNA; D015398:Signal Transduction; D023822:Tissue Engineering; D059467:Transcriptome
chemical_list: D035683:MicroRNAs; D020320:Oligoribonucleotides, Antisense; C546082:mirn21 microRNA, rat
keywords: Anti-miR; Cardiac hypertrophy; Engineered heart tissue; MicroRNA; Next generation sequencing