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Analysis and Validation of Differentially Expressed Ferroptosis-Related Genes in Regorafenib-Induced Cardiotoxicity
Siyuan Zhang1, Xueming Xu1, Zhangyi Li2
1The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150000, China.
Background:
Although tyrosine kinase inhibitors (TKIs) constitute a type of anticancer drugs, the underlying mechanisms of TKI-associated cardiotoxicity remain largely unknown. Ferroptosis is a regulated cell death form that implicated in several tumors' biological processes. Our objective was to probe into the differential expression of ferroptosis-related genes in regorafenib-induced cardiotoxicity through multiple bioinformatics analysis and validation.
Methods And Materials:
Four adult human cardiomyocyte cell lines treated with regorafenib were profiled using Gene Expression Omnibus (GEO) (GSE146096). Differentially expressed genes (DEGs) were identified using DESeq2 in R (V.3.6.3). Then, Gene Ontology (GO) Enrichment Analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) Enrichment Analysis, and Gene Set Enrichment Analysis (GSEA) were used to explore DEGs' bioinformatics functions and enriched pathways. We intersected DEGs with 259 ferroptosis-related genes from the FerrDb database. Finally, the mRNA levels of differentially expressed ferroptosis-related genes (DEFRGs) were validated in regorafenib-cultured cardiomyocytes to anticipate the link between DEFRGs and cardiotoxicity.
Results:
747,1127,773 and 969 DEGs were screened out in adult human cardiomyocyte lines A, B, D, and E, respectively. The mechanism by which REG promotes cardiotoxicity associated with ferroptosis may be regulated by PI3K-Akt, TGF-beta, and MAPK. GSEA demonstrated that REG can promote cardiotoxicity by suppressing genes and pathways encoding extracellular matrix and related proteins, oxidative phosphorylation, or ATF-2 transcription factor network. After overlapping DEGs with ferroptosis-related genes, we got seven DEFRGs and found that ATF3, MT1G, and PLIN2 were upregulated and DDIT4 was downregulated. The ROC curve demonstrated that these genes predict regorafenib-induced cardiotoxicity well.
Conclusion:
We identified four DEFRGs which may become potential predictors and participate in the regorafenib-induced cardiotoxicity. Our findings provide possibility that targeting these ferroptosis-related genes may be an alternative for clinical prevention and therapy of regorafenib-related cardiotoxicity.
Insights
Regorafenib-induced cardiotoxicity may involve ferroptosis, a cell death process. Researchers identified four differentially expressed ferroptosis-related genes (DEFRGs) that could predict and potentially treat this cardiotoxicity.
Area of Science:
- Cardiovascular Research
- Oncology
- Molecular Biology
Background:
- Tyrosine kinase inhibitors (TKIs) are anticancer drugs with largely unknown mechanisms of associated cardiotoxicity.
- Ferroptosis, a form of regulated cell death, is implicated in tumor biology.
- Understanding TKI-induced cardiotoxicity is crucial for patient safety.
Purpose of the Study:
- To investigate the differential expression of ferroptosis-related genes in regorafenib-induced cardiotoxicity.
- To identify potential biomarkers for regorafenib-induced cardiotoxicity.
- To explore therapeutic strategies targeting ferroptosis in TKI-related cardiotoxicity.
Main Methods:
- Bioinformatics analysis of gene expression data from regorafenib-treated human cardiomyocyte cell lines (GEO: GSE146096).
- Identification of differentially expressed genes (DEGs) using DESeq2.
- Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA) to explore gene functions and pathways.
- Intersection of DEGs with ferroptosis-related genes from the FerrDb database.
- Validation of differentially expressed ferroptosis-related genes (DEFRGs) mRNA levels.
Main Results:
- Multiple DEGs were identified across different cardiomyocyte cell lines.
- Regorafenib-induced cardiotoxicity may be regulated by pathways including PI3K-Akt, TGF-beta, and MAPK.
- GSEA indicated that regorafenib suppresses genes related to extracellular matrix, oxidative phosphorylation, and ATF-2 transcription factor network.
- Seven DEFRGs were identified, with ATF3, MT1G, and PLIN2 upregulated, and DDIT4 downregulated.
- Receiver Operating Characteristic (ROC) curve analysis demonstrated the predictive value of these genes for regorafenib-induced cardiotoxicity.
Conclusions:
- Four DEFRGs were identified as potential predictors and participants in regorafenib-induced cardiotoxicity.
- Targeting these ferroptosis-related genes may offer a novel approach for the clinical prevention and therapy of regorafenib-related cardiotoxicity.
- This study provides insights into the molecular mechanisms underlying TKI-associated cardiotoxicity.
