Identification Of Plasma Long Non-coding RNA-CoroMaker(IncRNA-CoroMaker) And Its Expression And Diagnostic Value In Coronary Atherosclerotic Heart Disease

Cardiovascular diseases, especially coronary artery disease(CAD), are still the major cause of death worldwide, causing a major socioeconomic burden world-wide. Although therapeutic methods such as medications, percutaneous coronary intervention, and coronary artery bypass surgery have improved the prognosis of CAD, the mortality rate remains high. Therefore, it is necessary to detect CAD in its earlier stage, especially before the development of left ventricular dysfunction[1,2]. The early identification of patients with CAD at high risk of adverse cardiovascular using circulating or imaging biomarkers may help in this regard[3]. However, available CAD biomarkers have limited risk prediction[4,5].Genome-wide analysis has identified that almost all of the human genome is transcribed, with a large amount of long non-coding RNAs(lncRNAs) [6,7]. LncRNAs, ranging over 200 nucleotides[8], have been found to be involved in specific physiological and pathological processes through epigenetic, transcriptional, or posttranscriptional regulatory mechanisms in a wide range of human diseases and disorders[9,10], for instance, cancers[11], and neurological disorders[12], Recently, several studies have shown that some lncRNAs are involved in the development of various types of cardiovascular dis eases[13-17], for example, heart failure[18-20], cardiac hypertrophy[21,22], cardiometabolic diseases[23] and myocardial infarction[24]. The plasma levels of some lnc RNAs, such as ANRIL, LincRNA-p21 and myocardial infarct-associated transcript-1(MIAT1), are markedly increased in atherosclerosis and may be important in its pathogenesis[25-27].LncRNAs are stable in plasma and other body fluids[28] and therefore, could serve as biomakers for some diseases. For example, a prostate specific lncRNA PCA3 in urine has been identified as the most specific biomarker for the detection of prostate cancer with higher specificity than the widely used PSA(prostate-specific antigen) test[28,29]. Other lncRNA biomarkers in plasma include H19 for gastric cancer[30], lncRNA HULC for hepatocellular carcinoma[31], and lncRNA LIPCAR for heart failure post myocardial infarction[18]. In the current study, lncRNAs were screened by microarray analysis and validated in different populations with or without CAD. We find that one plasma lncRNA and name lnc RNA –CoroMarker, is stable in plasma, exists in extracellular vesicle(EVs) mostly from monocytes, and is a sensitive and specific marker of CAD.Objective:lncRNAs have been reported to play a role in the pathogenesis of many cardiovascular diseases, but the research that circulating lncRNAs serviced as markers of cardiovascular disease is still in its infancy. In this study, high-throughput screening method is based on clinical specimens of patients with coronary heart disease, coronary heart disease aims to identify a new circulating lnc RNA and validated biomarkers to provide a new theoretical basis for early diagnosis of coronary heart disease.Method:1. RNA was extracted from clinical plasma samples for high-throughput mRNA and lncRNA microarray and analyzed test results by bioinformatics method.2. EVs were isolated from plasma by differential centrifugation.3. Collection and purification of monocytes with Peripheral blood.4. Total RNA was extracted from cells and plasma by TRIzol and purified.5. qRT-PCR was performed to test lncRNA expression.6. Receiver operating characteristic(ROC) curves and the area under the curve(AUC) were used to assess the feasibility of using plasma Coro Marker as a novel diagnosti c tool for the detection of CAD.7. Fisher criteria was used to determine if the predictive accuracy of Coro Marker as marker of CAD was improved with sex, hypertension, tobacco and alcohol use.8. Pearson correlation analysis was employed to verify the relationship between monocytes and plasma Coro Marker.Results:1. plasma lncRNA, CoroMarker, is stable in plasma, exists in extracellular vesicle(EVs) mostly from monocytes.2. Coro Marker expression was higher in CAD patients than in control sets. ROC curve analysis showed that Coro Marker might be a good candidate biomarker to predict CAD.3.The predictive value of Coro Marker was assessed, the corresponding optimal sensitivity was increased from 68.29% to 78.05%, and the specificity was slightly decreased from 91.89% to 86.49%, respectively.4. It was noted that the specificity of CoroMarker for CAD diagnosis, its plasma levels had increased spetificity compared with patients with other CVD.Conclusions:We conclude that Coro Marker is a stable, sensitive and specific biomarker for CAD..