Study On The Relationship Between Differential Expression Of MiRNA-mRNA And Hypertrophic Cardiomyopathy

Part Ⅰ Identification of key genes for hypertrophic cardiomyopathy based on bioinformatics methodsObjective To explore the molecular mechanisms and biomarkers of HCM and to identify important gene-related modules.Methods The gene expression profile of HCM from GEO and the genetic data from the MalaCards database with evidence for HCM were downloaded.After screening differentially expressed genes(DEG)by the online tool GEO2 R,functional enrichment analysis of DEG was performed using the clusterProfiler package in R,and GSEA gene set enrichment analysis was performed on all gene expression profile data.The DEG was combined with the Malacards database download gene to obtain the HCM-related gene(HCMRG).Then,use the STRING database and the CytoHubba and MCODE packages in Cytoscape to construct the protein-protein interaction(PPI)network of HCMRG and identify the central genes and related modules.Result 154 DEGs were obtained from GSE36961 dataset,and 225 HCMRGs were obtained by combining 72 genes in MalaCards database.A PPI network consisting of 214 nodes was constructed and 1151 edges wereconstructed.Three algorithms were used to identify each other.The first 16 central genes and 3 related modules.We found that TPM3,MYH6,PIM1,DUSP1,JAK2,STAT3,MYC,FOS,THBS1,SERPINE1,CCL2 may play a key role in HCM.Conclusion The results of this study may provide some guidance for further exploration of potential biomarkers for HCM patients.Part Ⅱ Analysis of miRNA-mRNA regulatory network in hypertrophic cardiomyopathyObjective To explore the molecular mechanisms and biomarkers of HCM and to construct a miRNA-mRNA regulatory network to guide the diagnosis and treatment of the disease in clinical practice.Methods Paired miRNA and mRNA expression profiles of myocardial tissue and normal myocardial tissue of HCM patients were collected from GEO,and differentially expressed miRNAs and genes were screened,and their functional characteristics and signal pathways were analyzed.Finally,the miRNA-mRNA regulatory network of HCM was constructed.Result Three miRNAs including miR-31-5p,miR-17-5p and miR-30 c and five risk genes including CCL2,FOS,JAK2,SERPINE1 and MYC played an important role in the miRNA-mRNA regulatory network.A number of miRNA target gene-associated risk pathways have been screened,possibly involving HCM diseases through interleukin-4 and interleukin-13 signaling(R-HSA-6785807)and positive regulation of cell death(GO: 0010942)process.Conclusion This study analyzed miRNA and mRNA expression profiles in HCM and screened three risk miRNAs(miR-31-5p,miR-17-5p,miR-30c)and five risk genes(CCL2,FOS,JAK2).,SERPINE1,MYC),in order to further explore the pathogenesis of HCM,providing a new way of thinking for thetargeted diagnosis and treatment of diseases.Part Ⅲ Verification of miRNA and mRNA chip results of HCMObjective To verify whether the expression of miRNAs and mRNAs at the gene level is consistent with the chip results.Methods In 5 HCM myocardium tissues and 5 normal myocardial tissues,3 candidate miRNAs(miR-31-5p,miR-17-5p,miR-30c)and 5 mRNAs(CCL2,FOS,JAK2,SERPINE1,MYC)were used to verify gene expression levels,and qRT-PCR experiments were used to compare the relative changes of miRNA and mRNA expression between the two groups,which laid a foundation for the molecular pathogenesis of HCM.Result After qRT-PCR experiments,the results showed that the expressions of 3 miRNAs and 5 mRNAs in HCM myocardial tissue and normal control myocardial tissue were statistically significant(P<0.05),and their expression differences and trends were related to the chip.The results are consistent.Conclusion Three miRNAs(miR-31-5p,miR-17-5p,miR-30c)and five mRNAs(CCL2,FOS,JAK2,SERPINE1,MYC)may be involved in the development of HCM.