Combined high-throughput library screening and next generation RNA sequencing uncover microRNAs controlling human cardiac fibroblast biology,Journal of Molecular and Cellular Cardiology

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Combined high-throughput library screening and next generation RNA sequencing uncover microRNAs controlling human cardiac fibroblast biology
Journal of Molecular and Cellular Cardiology ( IF 4.9 ) Pub Date : 2020-10-28 , DOI: 10.1016/j.yjmcc.2020.10.008
Katharina Schimmel ₁ , Stevan D Stojanović ₁ , Cheng-Kai Huang ₁ , Mira Jung ₁ , Martin H Meyer ₁ , Ke Xiao ₁ , Lea Grote-Levi ₁ , Christian Bär ₁ , Angelika Pfanne ₁ , Saskia Mitzka ₁ , Annette Just ₁ , Robert Geffers ₂ , Katharina Bock ₁ , Franziska Kenneweg ₁ , Felix Kleemiß ₁ , Christine S Falk ₃ , Jan Fiedler ₁ , Thomas Thum ₄

Affiliation

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.

中文翻译：

结合高通量文库筛选和下一代 RNA 测序揭示了控制人心脏成纤维细胞生物学的 microRNA

背景

心肌纤维化是心脏衰竭的一个标志，是全球最常见的死亡原因。近年来发现了几种控制心脏纤维化的 microRNA（miRNA、miR）；然而，目前还缺乏一种更全面的方法来识别参与纤维化的 miRNA。

方法和结果

功能性 miRNA 模拟库筛选应用于人心脏成纤维细胞 (HCF)，以鉴定诱导增殖的注释 miRNA。与此同时，在 HCF 受到增殖和静息刺激后进行 miRNA 深度测序，另外还能够发现新的 miRNA。深入的体外分析证实了选定的高度保守的 miRNA miR-20a-5p 和 miR-132-3p 的促纤维化性质。为了确定下游细胞通路及其在纤维化反应中的作用，通过合成 siRNA 调节注释的 miRNA 候选物的靶标。我们在此提供的证据表明，miR-20a-5p 和 miR-132-3p 对自噬的抑制和活性氧的解毒在机制水平上解释了它们的一些促纤维化性质。