National Health Commission Key Laboratory of Cardiovascular Regenerative Medicine, Heart Center of Henan Provincial People's Hospital, Central China Fuwai Hospital of Zhengzhou University, Fuwai Central China Cardiovascular Hospital and Central China Branch of National Center for Cardiovascular Diseases, Zhengzhou, China.
Department of Anesthesiology, Affiliated Drum Tower Hospital of Medical School of Nanjing University, Nanjing, China.
Clin Transl Med. 2023 May;13(5):e1258. doi: 10.1002/ctm2.1258.
Cardiac-resident or -enriched microRNAs (miRNAs) could be released into the bloodstream becoming circulating cardiac miRNAs, which are increasingly recognized as non-invasive and accessible biomarkers of multiple heart diseases. However, dilated cardiomyopathy (DCM)-associated circulating miRNAs (DACMs) and their roles in DCM pathogenesis remain largely unexplored.
Two human cohorts, consisting of healthy individuals and DCM patients, were enrolled for serum miRNA sequencing (10 vs. 10) and quantitative polymerase chain reaction validation (46 vs. 54), respectively. Rigorous screening strategy was enacted to define DACMs and their potentials for diagnosis. DCM mouse model, different sources of cardiomyocytes, adeno-associated virus 9 (AAV9), gene knockout, RNAscope miRNA in situ hybridization, mRFP-GFP-LC3B reporter, echocardiography and transmission electron microscopy were adopted for mechanistic explorations.
Serum miRNA sequencing revealed a unique expression pattern for DCM circulating miRNAs. DACMs miR-26a-5p, miR-30c-5p, miR-126-5p and miR-126-3p were found to be depleted in DCM circulation as well as heart tissues. Their expressions in circulation and heart tissues were proven to be correlated significantly, and a combination of these miRNAs was suggested potential values for DCM diagnosis. FOXO3, a predicted common target, was experimentally demonstrated to be co-repressed within cardiomyocytes by these DACMs except miR-26a-5p. Delivery of a combination of miR-30c-5p, miR-126-5p and miR-126-3p into the murine myocardium via AAV9 carrying an expression cassette driven by cTnT promoter, or cardiac-specific knockout of FOXO3 (Myh6-Cre , FOXO3 flox ) dramatically attenuated cardiac apoptosis and autophagy involved in DCM progression. Moreover, competitively disrupting the interplay between DACMs and FOXO3 mRNA by specifically introducing their interacting regions into murine myocardium crippled the cardioprotection of DACMs against DCM.
Circulating cardiac miRNA-FOXO3 axis plays a pivotal role in safeguarding against myocardial apoptosis and excessive autophagy in DCM development, which may provide serological cues for DCM non-invasive diagnosis and shed light on DCM pathogenesis and therapeutic targets.
心脏固有或富含的 microRNAs（miRNAs）可释放到血液中成为循环心脏 miRNAs，它们越来越被认为是多种心脏病的非侵入性和可及性生物标志物。然而，扩张型心肌病（DCM）相关的循环 miRNAs（DACMs）及其在 DCM 发病机制中的作用在很大程度上仍未得到探索。
两个人类队列，包括健康个体和 DCM 患者，分别进行了血清 miRNA 测序（10 对 10）和定量聚合酶链反应验证（46 对 54）。严格的筛选策略被制定来定义 DACMs 及其诊断潜力。采用 DCM 小鼠模型、不同来源的心肌细胞、腺相关病毒 9（AAV9）、基因敲除、RNAscope miRNA 原位杂交、mRFP-GFP-LC3B 报告基因、超声心动图和透射电子显微镜进行机制探索。
血清 miRNA 测序揭示了 DCM 循环 miRNAs 的独特表达模式。发现 miR-26a-5p、miR-30c-5p、miR-126-5p 和 miR-126-3p 在 DCM 循环和心脏组织中均被耗尽。它们在循环和心脏组织中的表达被证明是显著相关的，并且这些 miRNA 的组合被认为具有 DCM 诊断的潜在价值。FOXO3，一种预测的共同靶标，实验证明除 miR-26a-5p 外，这些 DACMs 在心肌细胞中共同受到抑制。通过携带受 cTnT 启动子驱动的表达盒的 AAV9 将 miR-30c-5p、miR-126-5p 和 miR-126-3p 的组合递送到小鼠心肌中，或心脏特异性敲除 FOXO3（Myh6-Cre，FOXO3 flox），可显著减弱 DCM 进展过程中涉及的心脏细胞凋亡和自噬。此外，通过特异性引入它们的相互作用区域到小鼠心肌中来破坏 DACMs 和 FOXO3 mRNA 之间的相互作用，削弱了 DACMs 对 DCM 的心脏保护作用。
循环心脏 miRNA-FOXO3 轴在保护 DCM 发展中心肌细胞凋亡和过度自噬方面发挥着关键作用，这可能为 DCM 的非侵入性诊断提供血清学线索，并为 DCM 的发病机制和治疗靶点提供启示。
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