Research On Nucleic Acids Biomarkers In Peripheral Blood Of Patients With Heart Failure Based On High-throughput Sequencing Platform

Objective Heart failure(HF)is a complex clinical syndrome caused by multiple factors and genetic changes,which is the result of gene regulation and environmental impact.At present,the early diagnosis and clinical treatment of heart failure are still not satisfactory.For heart failure,the symptoms and signs of patients have low sensitivity and specificity for clinical diagnosis,and the value of laboratory and imaging examination for early diagnosis and prognosis still needs to be improved.In some studies,peripheral blood mononuclear cells(PBMCs)were used as the media to identify biomarkers related to heart failure.However,the sample pretreatment process is complicated,which has limited its application in clinical laboratory diagnosis.In this study,peripheral blood cells(PBCs)of patients with heart failure were used as experimental materials to facilitate clinical detection.We used RNA-sequencing(RNA-seq),bioinformatics analysis and q RT-PCR to screen key differentially expressed genes(DEGs)from PBCs and further identified candidate nucleic acids biomarkers,which provides a new direction for clinical diagnosis of heart failure.Methods To better explore the characteristics of gene expression in patients with heart failure and identify the potential features of clinical diagnosis,we performed RNA-sequencing on PBCs samples from 8 patients with heart failure(HF group)and 10 healthy controls(HC group).In bioinformatics analysis section,we performed cluster analysis and co-expression analysis on these two groups.Fragments per kilobase of transcript per million fragments mapped(FPKM)was used to measure the expression of genes.The characteristics of gene expression profiles of PBCs in the HF group and the HC group were further described by screening the top 20 most abundant genes in the two groups.To identify significant DEGs,we compared the expression levels of all genes detected in each group.Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed to investigate the target DEGs of heart failure.The relative expression levels of significant DEGs of heart failure were validated by quantitative real-time PCR(q RT-PCR).Results The cluster analysis and correlation analysis showed that the HF group and the HC group were obviously divided into two groups and the gene expression of both groups was consistent as a whole.A total of 6123 genes were co-expressed in both groups after excluding the following FPKM >1 as baseline,while the HF and HC groups expressed 766 and 1349 genes,respectively.The results of differential expression analysis showed that 252 DEGs were detected in the two groups and gene expression patterns differed significantly between them.Among these DEGs,121 genes were up-regulated and 131 were down-regulated in the HF group.GO and KEGG analyses showed that these DEGs were mainly related to specific biological processes such as protein targeting transport,translation and macromolecular metabolism,as well as ribosome and phagosome related pathways.The top 10 differentially expressed protein-coding genes were validated using q RT-PCR.The q RT-PCR results of S100A9、FTL and EEF1A1 were highly consistent with the RNA-seq data.Area under curve(AUC)of receiver operating characteristic curve(ROC)was 0.800,indicating that these three genes may have the potential as nucleic acids biomarkers of heart failure.Conclusion S100A9,FTL and EEF1A1 can be used as candidate nucleic acids biomarkers in PBCs of patients with heart failure,which provides a new potential diagnostic value.Moreover,it is of great significance to further explore the signal pathways and biological functions related to heart failure.