Coronary Heart Disease LncRNA Expression Profile Analysis, Biomarker Identification And Preliminary Functional Research

Background and Objective:Dysregulation of long non-coding RNAs(IncRNAs)has been proven to be involved in the pathogenesis of coronary artery disease(CAD).However,it remains to be extensively explored.Thus,we studied expression patterns,biological functions,and diagnostic value of lncRNAs in CAD.Our studywould be helpful to provide new insights for the prevention,diagnosis and treatment of CAD.Methods:Using microarray,we performed the transcriptome-wide lncRNAs and mRNAs expression profile in peripheral blood mononuclear cells(PBMCs)of 93 CAD patients and 48 healthy controls.Gene Ontology(GO)and pathway analysis for differentially expressed mRNAs was used to investigate underlying biological associations of differentially expressed IncRNAs and establish path-net todepict interactions of significant pathways.Quantitative Realtime-PCR(qRT-PCR)was used to validate selected IncRNAs in 412 CAD patients and 295 healthy controls.Receiver operating characteristic(ROC)curve analysis was performed to evaluate whether lncRNAs could be used to diagnose CAD patients.Finally,the functional significance of several validated IncRNAswas determined in THP-1-derived macrophages.The knockdown of AS03973 by specific siRNAs and the overexpression by adenovirus vector in HUVECs were performed,respectively.qRT-PCR,Western Blot,ELISA and Flow cytometry were performed to test the mRNA expression and protein expression of inflammatory cytokine genes and adhesion molecule genes,respectively.Results:We identified 1,210 IncRNAs and 890 mRNAs differentially expressed from the expression profile and validated selected seven lncRNAs.Two novel IncRNA biomarkers,ENST00000444488.1 and ucO10yfd.1,together with CAD risk factors had the better predictive performance for discriminating CAD patients from healthy controls,and ENST00000444488.1 could diagnose acute myocardial infarction(AMI)patients from non-AMI patients.The knockdown of ENST00000444488.1,uc010yfd.11 AS03973 and ENST00000602558.1 affected the expression of inflammation-related genes and their nearby genes in THP-1-derived macrophages.Compared with the negative control,knockdownof AS03973 significantly reduced the expression oflL-6 and ICAM-lwhile the overexpression of AS03973 significantly increased the expression of IL-6 and ICAM-1.Conclusion:We offered a transcriptome-wide overview of aberrantly expressed IncRNAs in CAD patients,and identified two novel IncRNA biomarkers for diagnosing CAD.Loss of some validated IncRNAs regulated the expression of inflammation-related genes and their nearby genes in macrophages.LncRNA AS03973 could regulate the expression of IL-6,ICAM-1 in endothelial cells.From the above,the present study demonstrates that IncRNA ENST00000444488.1anduc010yfd.1 could be the new biomarker to diagnose CAD.The result of regulation expression of inflammation-related genes and nearby genes by indicated IncRNAs is helpful to provide new clues for seeking the target genes of CAD.The results suggested that AS03973 might be involved in the endothelial dysfunction and the emergence of atherosclerosis through regulating inflammatory cytokine and intercellular adhesion molecule,which could offer more evidence to investigate the relationship between IncRNA and CAD and support to the further task of the function research for CAD.