Methods: The GSE66360 dataset was used to identify Differentially Expressed Genes (DEGs), and pathway differences were analyzed via Gene Set Enrichment Analysis (GSEA). The core modular genes were selected after clustering gene modules related to cuproptosis and mitophagy using Weighted Gene Co-Expression Network Analysis (WGCNA). LASSO regression and random forest were employed for feature selection. Immune microenvironment profiling was conducted to identify its potential therapeutic targets.

Results: The DEGs were associated with cell cycle, glycolysis, and the inflammatory signaling pathway. The dark green module identified by WGCNA was correlated with both cuproptosis and mitophagy and was enriched in oxidative phosphorylation and immune processes. A three-gene diagnostic model was established and found to be upregulated in MI and correlated with immune infiltration. Molecular docking analysis indicated that Corbadrine and Dicyclomine are potential therapeutic agents targeting CD55.

Discussion: This study highlights the broader implications of linking cuproptosis and mitophagy in MI, proposing a novel perspective on mitochondrial dysfunction as a central hub connecting metabolic stress, immune dysregulation, and cell death. The identified core genes, S100A8, CD55, and GPCPD1, not only serve as potential diagnostic markers but may represent functional nodes at the intersection of copper-dependent cell death and mitochondrial quality control.

Conclusion: The core genes (GPCPD1, S100A8, and CD55) not only functioned as potential diagnostic markers but also served as functional nodes in copper-dependent cell death, contributing to MI treatment.

Methods: Following myocardial infarction (MI) induction, we performed transcriptome sequencing of CFs isolated from R26-LSL-TFEB+/+; Acta2-cre (n = 3) and R26-LSL-TFEB+/+ (n = 3) mice. Differential gene expression and functional enrichment analyses were conducted using R software. The binding between TFEB and Thrombospondin-1 (Thbs1) was validated by ChIP-qPCR assay. CFs were extracted from adult mice in the R26-LSL-TFEB+/+; Acta2-cre and R26-LSL-TFEB+/+ groups. Protein expressions of integrin, CD47, CD36, Thbs1, p-paxillin, vinculin, P-FAK, and α- SMA were detected by Western blot. Cell migration was assessed by the wound healing and Transwell assays.

Results: TFEB modulated the expression of a broad spectrum of RNAs associated with the transformation of CFs. Pathway analysis revealed significant enrichment in pathways related to extracellular matrix (ECM) receptor interaction and focal adhesion (FA). Notably, both mRNA and protein levels of Thbs1 were markedly elevated in TFEB-overexpressing CFs. Integrated computational prediction and chromatin immunoprecipitation assays identified that TFEB directly bound to the promoter region of Thbs1. This binding was associated with downstream modulation of its receptor network and a concomitant reduction in FA complex activation at the protein level. These findings positioned Thbs1 as a key transcriptional target through which TFEB regulated ECM-related signaling and cellular adhesion dynamics in CFs.

Discussion: The current findings showed that the modulation of Thbs1 and associated FA signaling was a mechanism through which TFEB overexpression exerted its anti-fibrotic effects on CFs. This highlighted the TFEB-Thbs1 axis as a potential novel target for developing therapeutic strategies to mitigate cardiac fibrosis.

Conclusions: This study suggested that the protective effect of TFEB against MI injury was associated with the Thbs1/FA signaling pathway, providing a novel potential therapeutic target for cardiac fibrosis.