A phenotypic screen to identify hypertrophy-modulating microRNAs in primary cardiomyocytes
A phenotypic screen to identify hypertrophy-modulating microRNAs in primary cardiomyocytes
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DOI：
10.1016/j.yjmcc.2011.07.010
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发表时间：
2012-01-01
影响因子：
5
通讯作者：
Engelhardt, Stefan
中科院分区：
文献类型：
--
作者：
Jentzsch, Claudia;Leierseder, Simon;Engelhardt, Stefan
MicroRNAs(MiRNAs)是一种控制互补靶向mRNAs表达的非编码小RNA。越来越多的miRNAs与心脏疾病的发病机制有关，主要不是基于功能数据，而是因为观察到它们在病变心肌中调节失调。因此，我们对大多数miRNAs潜在的心脏作用的了解是有限的。在这里，我们报告了一种允许同时分析数百个分子对原代大鼠心肌细胞表型影响的分析格式的发展。使用自动显微镜和边缘检测算法，该分析实现了高重复性和作为关键参数的心肌细胞大小的稳健评估。筛选合成的miRNAs文库发现了几个以前没有被认为是促进或抗肥大的miRNAs。从中筛选出9个miRNAs，证实了miR-22、miR-30c、miR-30d的促肥大作用。MiR-212、miR-365以及miR-27a、miR-27b和miR-133a的抗肥大作用。对原代心肌细胞中促进肥大的miRNAs表达水平的定量分析表明，单个miRNAs的表型效应与其表达水平之间的相关性相当低。这种检测方法可以自动测定原代心肌细胞的大小，并可以鉴定出一组能够调节心肌细胞肥大的miRNAs。阐明它们的作用机制将有助于深入了解心肌细胞肥大反应的机制。本文是题为《可能的社论》的特刊的一部分。(C)2011爱思唯尔有限公司。保留所有权利。
MicroRNAs (miRNAs) are small non-coding RNAs that control expression of complementary target mRNAs. A growing number of miRNAs has been implicated in the pathogenesis of cardiac diseases, mostly based not on functional data, but on the observation that they are dysregulated in diseased myocardium. Consequently, our knowledge regarding a potential cardiac role of the majority of miRNAs is limited. Here, we report the development of an assay format that allows the simultaneous analysis of several hundred molecules with regard to their phenotypic effect on primary rat cardiomyocytes. Using automated microscopy and an edge detection algorithm, this assay achieved high reproducibility and a robust assessment of cardiomyocyte size as a key parameter. Screening a library of synthetic miRNAs revealed several miRNAs previously not recognized as pro- or anti-hypertrophic. Out of these, we selected nine miRNAs and confirmed the pro-hypertrophic potential of miR-22, miR-30c, miR-30d. miR-212, miR-365 and the anti-hypertrophic potential of miR-27a, miR-27b and miR-133a. Quantitative analysis of the expression level of pro-hypertrophic miRNAs in primary cardiomyocytes indicated a rather low level of correlation of the phenotypic effects of individual miRNAs and their expression level. This assay allows the automated determination of cell size in primary cardiomyocytes and permitted the identification of a set of miRNAs capable of regulating cardiomyocyte hypertrophy. Elucidating their mechanism of action should provide insight into mechanisms underlying the cardiomyocyte hypertrophic response. This article is part of a Special Issue entitled 'Possible Editorial'. (C) 2011 Elsevier Ltd. All rights reserved.