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

Chen Tianlong; Qiao Bokang; Jia Lixin; Du Jie

doi:10.3760/j.issn.1673-4777.2025.11.002

Abstract：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,respec-tively.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.