2025 年第 11 期 第 20 卷

剪切体抑制剂联合表观遗传重塑药物促进心脏重编程的转录组学分析

Transcriptomic analysis of spliceosome inhibitor combined with epigenetic remodeling drugs to promote cardiac reprogramming

作者：陈天龙乔博康贾立昕杜杰

英文作者：Chen Tianlong Qiao Bokang Jia Lixin Du Jie

单位：首都医科大学附属北京安贞医院转化医学中心北京市心肺血管疾病研究所，北京100029

英文单位：Translational Medicine Center Beijing Anzhen Hospital Capital Medical University Beijing Institute of Heart Lung and Blood Vessel Diseases Beijing 100029 China

关键词：心肌重编程；转分化；剪切体抑制剂；表观调控小分子药物；转录组测序

英文关键词：

• 摘要：

• 目的 探究剪切体抑制剂联合表观遗传重塑药物在心肌重编程中的作用。方法 将出生后1 d小鼠（C57BL/6J）分为二甲基亚砜（DMSO）组、CRFV（一组转分化的核心小分子药物，简称为CRFV）组及CRFV+PLaB（一种剪切体抑制剂）组，每组3只。提取小鼠心脏成纤维细胞后进行转录组测序；基于P-value以及表达量改变分析差异基因，使用实时定量聚合酶链反应对部分差异基因进行验证；分别对显著上调和下调的基因进行功能富集分析和信号通路富集分析揭示药物处理对成纤维细胞的功能调控。结果 对原代心脏成纤维细胞进行联合药物7 d处理，经转录组测序分析发现心脏相关基因Tnnt2、Actn2、Myh6、Gata4均显著升高，成纤维细胞相关基因Col1a1、Col1a2、S100a4均显著下降。富集分析结果表明，上调基因与肌动蛋白丝形成、细胞形态发生密切相关；而下调基因则与细胞外基质形成、心肌肥大相关；实时荧光定量结果表明成纤维细胞特异性标志物(Col1a1、Col1a2)表达降低，心肌细胞特异性标志物(Tnnt2、Actn2、Myh6)表达显著升高。结论 剪切体抑制剂和表观遗传重塑的药物可促进心脏成纤维细胞重编程为心肌样细胞。

• Objective To explore the role of spliceosome inhibitor combined with epigenetic remodeling drugs in myocardial reprogramming. Methods One-day-old C57BL/6J mice were divided into dimethyl sulfoxide (DMSO) group, CRFV (a core small-molecule drug for transdifferentiation, abbreviated as CRFV) group, and CRFV + PLaB (a spliceosome inhibitor) group, with 3 mice in each group. Mouse cardiac fibroblasts were extracted for transcriptome sequencing. The differentially expressed genes were analyzed based on P-value and expression change, and some differentially expressed genes were verified by real-time quantitative polymerase chain reaction. Functional enrichment analysis and signaling pathway enrichment analysis of significantly up-regulated and down-regulated genes were performed to reveal the functional regulation of drug treatment on fibroblasts, respectively. Results Transcriptome sequencing analysis showed that the heart-related genes Tnnt2, Actn2, Myh6, and Gata4 were significantly increased, and the fibroblast-related genes Col1a1, Col1a2, and S100a4 were significantly decreased in primary cardiac fibroblasts treated with combined drugs for 7 d. Enrichment analysis showed that up-regulated genes were closely related to actin filament formation and cell morphogenesis. Down-regulated genes were related to extracellular matrix formation and cardiac hypertrophy. Real-time fluorescence quantitative results showed that the expression of fibroblast-specific markers (Col1a1 and Col1a2) was decreased, and the expression of cardiomyocyte-specific markers (Tnnt2, Actn2, and Myh6) was significantly increased. Conclusion Spliceosome inhibitor and epigenetic remodeling drugs promote the reprogramming of cardiac fibroblasts into cardiomyocyte-like cells.