Extracellular Matrix Secretion by Cardiac Fibroblasts Role of MicroRNA-29b and MicroRNA-30c
Extracellular Matrix Secretion by Cardiac Fibroblasts Role of MicroRNA-29b and MicroRNA-30c
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DOI：
10.1161/circresaha.113.302400
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发表时间：
2013-10-25
影响因子：
20.1
通讯作者：
Mayr, Manuel
中科院分区：
文献类型：
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作者：
Abonnenc, Melanie;Nabeebaccus, Adam A.;Mayr, Manuel
基本原理：目的：研究miR-29 b和miR-30 c对心肌成纤维细胞分泌细胞外基质的影响，并比较其蛋白质组学差异。miR-29 b靶向参与纤维化的蛋白质干部，包括多种胶原蛋白、基质金属蛋白酶和白血病抑制因子、胰岛素样生长因子1和五聚蛋白3，3预测miR-29 b的靶点。miR-29 b还减弱了心脏成纤维细胞对转化生长因子-β的反应。相比之下，miR-30 c对细胞外基质产生的影响很小，但对白血病抑制因子和胰岛素样生长因子1的影响相反。两种miRNAs均间接影响心肌细胞。转染前体miR-29 b后，心脏成纤维细胞的条件培养基失去了支持大鼠心室肌细胞粘附的能力，并导致心肌细胞蛋白（α-辅肌动蛋白、心肌肌球蛋白结合蛋白C和心肌肌钙蛋白I）显著减少。类似地，源自小鼠胚胎干细胞的心肌细胞在pre-miR-29条件培养基下萎缩，而pre-miR-30 c条件培养基具有促肥大作用。在病理性而非生理性肥大的小鼠模型中，miR-29 a、miR-29 c和miR-30 c的水平显著降低，但miR-29 b没有。治疗与miR-29 b的miRNAs诱导过度纤维化后，主动脉缩窄没有明显恶化的心脏functions.Conclusions：我们的蛋白质组学分析揭示了新的分子靶点的miRNAs与纤维化的心脏表型。这种全面的筛选方法对于确定miRNAs在心血管疾病中的协同作用至关重要。
Rationale: MicroRNAs (miRNAs), in particular miR-29b and miR-30c, have been implicated as important regulators of cardiac fibrosis.Objective: To perform a proteomics comparison of miRNA effects on extracellular matrix secretion by cardiac fibroblasts.Methods and Results: Mouse cardiac fibroblasts were transfected with pre-/anti-miR of miR-29b and miR-30c, and their conditioned medium was analyzed by mass spectrometry. miR-29b targeted a cadre of proteins involved in fibrosis, including multiple collagens, matrix metalloproteinases, and leukemia inhibitory factor, insulin-like growth factor 1, and pentraxin 3, 3 predicted targets of miR-29b. miR-29b also attenuated the cardiac fibroblast response to transforming growth factor-beta. In contrast, miR-30c had little effect on extracellular matrix production but opposite effects regarding leukemia inhibitory factor and insulin-like growth factor 1. Both miRNAs indirectly affected cardiac myocytes. On transfection with pre-miR-29b, the conditioned medium of cardiac fibroblasts lost its ability to support adhesion of rat ventricular myocytes and led to a significant reduction of cardiac myocyte proteins (a-actinin, cardiac myosin-binding protein C, and cardiac troponin I). Similarly, cardiomyocytes derived from mouse embryonic stem cells atrophied under pre-miR-29 conditioned medium, whereas pre-miR-30c conditioned medium had a prohypertrophic effect. Levels of miR-29a, miR-29c, and miR-30c, but not miR-29b, were significantly reduced in a mouse model of pathological but not physiological hypertrophy. Treatment with antagomiRs to miR-29b induced excess fibrosis after aortic constriction without overt deterioration in cardiac function.Conclusions: Our proteomic analysis revealed novel molecular targets of miRNAs that are linked to a fibrogenic cardiac phenotype. Such comprehensive screening methods are essential to define the concerted actions of miRNAs in cardiovascular disease.