Identification of Ferroptosis-Related Genes in Heart Failure Induced by Transverse Aortic Constriction

Abstract

Background

Heart failure (HF) is a common clinical syndrome due to ventricular dysfunction and is a major cause of mortality worldwide. Ferroptosis, marked by excessive iron-dependent lipid peroxidation, is closely related to HF. Therefore, the purpose of this study is to explore and validate ferroptosis-related markers in HF by bioinformatics analysis and animal experiments validation.

Materials and Methods

The gene expression profiles (GSE36074) of murine transverse aortic constriction (TAC) were obtained from the Gene Expression Omnibus (GEO); From the FerrDb database, ferroptosis-related genes (FRGs) were identified. Using GEO2R, differential expressed genes (DEGs) were screened. An overlapping analysis was conducted among DEGs and FRGs to identify ferroptosis-related DEGs (FRDEGs). We then performed clustering, functional enrichment analysis, and protein-protein interaction (PPI) analyses. In addition, the key FRDEGs were extracted by cytoHubba plugin and the networks of transcription factors (TFs)-key FRDEGs and microRNA-key FRDEGs were constructed. Lastly, the key FRDEGs were carried by quantitative reverse transcription PCR (RT-qPCR) and immunohistochemistry (IHC).

Results

Fifty-nine FRGs showing significantly different expression were identified from a total of 1918 DEGs in mice heart by transverse aortic constriction. GO and KEGG functional enrichment analysis revealed that these 59 ferroptosis-related DEGs mostly associated with positive regulation of apoptotic process, FoxO signaling pathway, VEGF signaling pathway, Apoptosis, Ferroptosis. Five key FRDEGs (Mapk14, Hif1a, Ddit3, Tlr4 and Ptgs2) were identified using PPI networks; Based on TFs-key FRDEGs networks, we found that Mapk14, Hif1a, Tlr4 and Ptgs2 were regulated by 3, 4, 5, and 29 TFs, respectively; however, Ddit3 was not regulated by any TF; By analyzing the miRNA–key FRDEGs networks, we found that 39, 74, 11, 28, and 18 miRNAs targets regulate the expression of Mapk14, Hif1a, Ddit3, Tlr4 and Ptgs2, respectively. Lastly, five key FRDEGs were validated at the mRNA and protein levels by RT-qPCR and IHC, which were in line with our bioinformatics analysis.

Conclusion

Our findings reveal that Mapk14, Hif1a, Ddit3, Tlr4 and Ptgs2 may be involved in the development of HF through regulating ferroptosis and as potential targets for HF.

Keywords:

• heart failure
• transverse aortic constriction
• ferroptosis
• bioinformatics
• key genes

Abbreviations

HF, Heart failure; TAC, Transverse aortic constriction; FRDEGs, Ferroptosis-related differential expressed genes; PPI, Protein-protein interaction; RT-qPCR, Quantitative reverse transcription PCR; IHC, Immunohistochemistry; CVDs, Cardiovascular diseases.

Data Sharing Statement

The data is available in NCBI GEO, accession number: GSE36074 and ementary material.

Ethics Statement

In this study, there does not involve human research, infringe on the privacy of human, and include any human biological samples. Therefore, the ethics were approval waived by Medical College, Yangzhou University.

Acknowledgments

We are indebted to Prof. Skrbic B who shared their data in GEO.

Disclosure

The authors report no conflicts of interest in this work.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China (Grant No.81800250), China Postdoctoral Science Foundation (No.2022M711417), Yangzhou science and technology plan social development project (No. SSF2023000133).