J Gene Med. 2026 Aug;28(8):e70102. doi: 10.1002/jgm.70102.
ABSTRACT
BACKGROUND: Myocardial ischemia-reperfusion injury (MIRI) remains a major clinical problem, and its regulatory mechanisms are not fully defined. This study aimed to identify key MIRI-related targets and clarify their roles in cellular and animal models.
METHODS: Differentially expressed genes in MIRI were screened using the GEO dataset GSE6381, and core genes were identified through protein-protein interaction analysis and weighted gene co-expression network analysis. External datasets GSE249812 and GSE123342 were used to validate core gene expression. Transcription factors regulating cystic fibrosis transmembrane conductance regulator (CFTR) were predicted using the KnockTF database. In AC16 cells, an oxygen-glucose deprivation/reoxygenation (OGD/OGR) model was established. Microphthalmia-associated transcription factor (MITF) was knocked down or overexpressed, CFTR was silenced or overexpressed, and cell viability, proliferation, apoptosis, and NO/L-arginine/citrulline levels were assessed. The MITF-CFTR interaction was examined by dual-luciferase assay and chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR). A rat ischemia-reperfusion (I/R) model was used to evaluate the effects of MITF silencing and CFTR rescue on cardiac injury.
RESULTS: A total of 42 core genes related to MIRI were identified. CFTR showed consistent upregulation across datasets and was linked to pathways including arginine biosynthesis. CFTR-related transcription factors included MITF, TP53, and STAT3, but ChIP-qPCR showed detectable enrichment only for MITF at the CFTR promoter. In AC16 cells, CFTR protein increased during early reoxygenation. OGD/OGR elevated CFTR and MITF, while MITF knockdown or CFTR silencing further reduced cell viability and proliferation, increased apoptosis, and was accompanied by lower NO and citrulline levels and higher L-arginine levels. MITF overexpression showed the opposite trend, and CFTR overexpression partially reversed the effects of MITF deficiency. In vivo, MITF knockdown aggravated cardiac dysfunction, increased CK and CK-MB levels, worsened tissue injury, and elevated cleaved caspase-3 after I/R, whereas CFTR overexpression partly improved these changes.
CONCLUSIONS: MITF regulates CFTR expression in MIRI, and disruption of the MITF-CFTR axis is associated with aggravated injury in cellular and animal models. Restoring CFTR partially mitigates MITF deficiency-related damage. These findings support the involvement of MITF-CFTR regulation in MIRI and provide a basis for further mechanistic investigation.
PMID:42562793 | DOI:10.1002/jgm.70102