The Clinical Value Of Urine MiRNA In The Early Diagnosis Of Kawasaki Disease

Part ?.Differences in miRNA expression between blood and urine in acute phase of children with Kawasaki diseaseObjective:To understand the expression of miRNA in platelet and the expression of miRNA in urine in the acute phase of Kawasaki disease(KD),compare the difference in miRNA composition,content and expression level between the platelet and urine,to verify urine miRNA detection and correlation of Kawasaki disease diagnosis.Methods:10 children with Kawasaki disease were admitted to the Children's Hospital of Suzhou University,and 10 children with other febrile diseases were enrolled in the same period as the control group.Collect clinical data and keep samples of peripheral blood and urine in the acute phase.The expression of miRNA in peripheral blood platelets and the expression of miRNA in urine were detected by high-throughput sequencing technology,and to compare the differential expression between the two groups.Results:(1)Through high-throughput sequencing technology,it was found that children with KD had 46 differentially expressed miRNAs in acute phase platelets compared with other febrile diseases,of which 10 were significantly up-regulated by more than 1.5 times and 11 were significantly down-regulated.(2)The miRNAs in the peripheral blood and urine of the Kawasaki group and the fever control group showed that there were 37 miRNAs specific to platelets,accounting for 20.4%,and 23 miRNAs specific to urine,accounting for 12.7%,and miRNAs shared by platelets and urine,there are 121,the overall agreement is 66.8%,close to 70%.Conclusion:The content of miRNA in urine is lower than that in blood,but the size and distribution of miRNA fragments extracted from urine are basically the same as those in blood,and the integrity of RNA is also good.The composition of peripheral blood miRNA and urine miRNA,the degree of coincidence is high.Urine miRNA can be used for Kawasaki disease prediction.Part ?.Urine miRNA predicts the risk of Kawasaki disease in children with high feverObjective:To find miRNAs specifically expressed in the urine of children with acute Kawasaki disease(KD),as a biomarker,to construct a predictive model for the risk of Kawasaki disease in children with high fever.Methods:10 children with Kawasaki disease were admitted to the Children's Hospital of Suzhou University,and 10 children with other febrile diseases were enrolled in the same period as the control group,the peripheral blood and urine samples of acute phase were collected for specific miRNA in urine(biomarkers)search.In addition,10 other KD children and 8 other febrile diseases were selected,the urine samples of acute phase were collected for verification of the auxiliary diagnosis of Kawasaki disease model.High-throughput sequencing technology was used to detect miRNAs specifically expressed in urine,and the Linear SVM machine learning analysis model was used to rank the importance of the detected miRNAs for distinguishing children with Kawasaki disease and fever control groups.The two top miRNAs were used as urine biomarkers and were validated in an additional 18 samples.Results:(1)There were 12 miRNAs significantly different in the urine between the Kawasaki group and other febrile disease control groups,Among them,hsa-miR-223-3p,hsa-miR-7-5p,hsa-miR-99b-5p,hsa-miR-423-5p,hsa-miR-320a,hsa-miR-182-5p was significantly up-regulated in the urine of patients with Kawasaki disease of acute phase(P<0.05,FDR<0.05),and hsa-let-7g-5p,hsa-let-7i-5p,hsa-miR-199b-3p,hsa-miR-199a-3p,hsa-miR-27a-3p,hsa-miR-25-3p was significantly down-regulation in the urine of patients with Kawasaki disease of acute phase(P<0.05,FDR<0.05).(2)The top two most important ones are the hsa-miR-223-3p and hsa-let-7g-5p,which are ranked by the Linear SVM machine learning analysis model.Using the expression levels of miR-223-3p and hsa-let-7g-5p,18 samples were validated by the Linear SVM analysis model,draw ROC curve,it was found that only miR-223-3p and has-let-7g-5p were used as training characteristics,the AUC could reach 0.96,the specificity was 91%,and the sensitivity was 97%.Conclusion:There are 12 miRNAs characteristically expressed in the urine of acute Kawasaki disease.Among them,miR-223-3p was significantly up-regulated in the urine of Kawasaki disease,while hsa-let-7g-5p was significantly down-regulated.These two miRNAs were used to distinguish the most diagnostic value of children with Kawasaki disease and fever control group.It can be used as one of the urine biomarker for early diagnosis of Kawasaki disease.