Transcriptome High-throughput Sequencing Analyzes Potential Biomarkers For White Matter Lesions

BackgroundHypertension is confirmed to be one of the major risk factors of leukoaraiosis(WMLS).However,not all hypertensive patients showed WMLs,and some hypertensive patients never showed white matter damage.The pathogenesis of leukoaraiosis is not completely understood and there is no reliable indicator for the early diagnosis of WMLS in the hypertensive population.Long non-coding RNAs(lncRNAs)may have important significance for WMLs,which is an important reason of cognitive impairment.Using this as a starting point,we performed transcriptome high-throughput sequencing in hypertensive patients with or without WMLs to analyze the potential biological markers leading to the occurrence of WMLs.ObjectiveTo analyze the potential biomarker for the diagnosis in WMLs patients with hypertension and to further explore the biological process and possible mechanism of WMLs.Methods1.Subjects were divided into WMLs group(Hypertensive patients with WMLs)and control group(Hypertensive patients without WMLs).Gene expression profiles of the two groups were analyzed by transcriptome high-throughput sequencing.2.GO analysis was performed on differentially expressed(DE)mRNAs.The potential functions of DE mRNA in the WMLs group compared with the control group was explored.3.According to cis/trans analysis,lncRNAs related to DE mRNA were traced to the source and the traceable lncrnas were verified by qPCR to obtain the potential biological markers of WMLs4.The sample size was further expanded to verify the stability of the potential biological marker,and the predictive power of the marker was evaluated by ROC curve.Results1.The gene expression profiles were analyzed and heat maps were made,indicating that the patients in the two groups could be clustered through the mRNA expression profiles.2.Through GO analysis,DE mRNA was enriched into the biological processes related to WMLs.Combined with previous studies,there were 4 biological processes that may be related to WMLs(including reverse cholesterol transport,regulation of cytokine secretion,inflammatory cell apoptotic process and interleukin-6 secretion).3.According to cis/trans analysis,15 lncmas related to mRNA were traceable to the first 6 lncRNAs with significant statistical significance were selected for qPCR verification(including CTD-3064M3.4,SLC9A9-AS2,LINC01013,AC011747.3,CTD-2162K18.4,CTC-246B 18.10).4.After expanding the sample size,qPCR verification showed that only CTD-2162k 18.4 might be a potential biological marker of WMLs.The area under the ROC curve of CTD-2162kl8.4 was 0.9062(95%IC:0.8508-0.9615).When the threshold value was 0.01654,the separation effect between the WMLs group and the control group was the best(sensitivity was 78.00%,specificity was 90.00%).Conclusions1.The results of the present study manifested that the lncRNA may have a significant role in the pathogenesis of WMLs.And it may be a novel target for further research.2.Through transcriptome high-throughput sequencing analysis and validation,it was found that CTD-2162k18.4 may be a potential biological marker for patients with hypertension accompanied by WMLs.3.WMLs may be related to the reverse cholesterol transport,regulation of cytokine secretion,inflammatory cell apoptotic process and interleukin-6 secretion,and dyslipidemia and inflammatory response may play an important role in the pathogenesis of WMLs.As the relationship between IncRNAs and WMLS is just beginning to be unraveled,their specific mechanisms require further investigation.