Identification And Functional Analysis Of Circulating MiRNAs And Their Target Genes In Myasthenia Gravis Patients

Objective: To analysis the expression profile of miRNA in plasma of patients with Myasthenia Gravis and explore potential biomarkers in blood.To screen the differential expressed circulating miRNA,explore the pathogenesis of myasthenia gravis at the molecular level and provide new ideas for diagnosis and treatment in the future.Method:1.Plasma samples of experimental group(patients with myasthenia gravis)and the recruited healthy control group were collected from the department of Neurology,Second Xiangya Hospital,Central South University.2.miRNA expression profiles in plasma of experimental group and control group were analyzed by second-generation sequencing.3.An independent cohort validation experiment was conducted for significantly differential expressed miRNAs.Quantitative analysis was performed by real-time fluorescence quantitative assay(RT-q PCR),and ROC curve was used to evaluate the possibility of plasma miRNA as potential biomarkers for myasthenia gravis.4.Circulating miRNA related to myasthenia gravis were further screened from Pub Med,Web of Science and Google Scholar databases,and target Gene prediction and GO,PPI and KEGG pathways analysis were carried out through online database.Transcriptome data from the GEO database were analyzed to assess hub-genes of myasthenia gravis.Results:1.Compared with normal,total 30 differentially expressed miRNA were screened by sequencing,including 13 up-regulated miRNAs and 17down-regulated miRNA,the criteria for screening is P<0.05,| log FC | >1.2.The down-regulated miR-196a-5p was affirmed by RT-q PCR,and the results were consistent with the sequence.ROC curve analysis showed that the AUC value of miR-196a-5p was 0.7222(sensitivity,66.7%;specificity,66.7%,95%CI: 0.5575 to 0.8870).3.A total of 16 differentially expressed miRNAs in serum or plasma of MG patients were screened from article databases which were affirmed by RT-q PCR and the sample size of validation set is more than 10.4.Differentially expressed miRNA are considered to participate in cancer related pathway,hepatitis B related pathway,MAPK and PI3K-Akt signal pathway by GO,KEGG pathway analysis and PPI interaction analysis.5.TP53,AKT1,STAT3,CCND1,KRAS,HIF1 A,VEGFA,ERBB2,SMAD2 and PTEN which targeted by miR-30e-5p,miR-150-5p,miR-133a-3p,miR-20 b,miR-21-5p,miR-19b-3p,miR-27a-3p,miR-140-5p,miR-185-5p and miR-15b-5p are top ten hub genes of myasthenia gravis.Conclusion:1.miR-196a-5p is down-regulated in plasma of patients with myasthenia gravis and is expected to be a potential biomarker for myasthenia gravis.2.TP53,AKT1,STAT3,CCND1,KRAS,HIF1 A,VEGFA,ERBB2,SMAD2 and PTEN are hub gene of myasthenia gravis.Circulating miR-30e-5p,miR-150-5p,miR-133a-3p,miR-20 b,miR-21-5p,miR-19b-3p,miR-27a-3p,miR-140-5p,miR-185-5p and miR-15b-5p target hub genes and may play important regulatory in the pathological process of myasthenia gravis by tumor related pathway,hepatitis B related pathway,MAPK and PI3K-Akt signaling pathway.