Analysis Of The Expression Level Of Plasma Exosomes Microrna In Patients With Intracranial Aneurysms

Objective The aim of this study was to investigate the expression levels of plasma exosomal microRNA(miRNA)in patients with intracranial aneurysm(IA)to find potential biomarkers that are closely related to the occurrence,development,and rupture of IA.Methods A total of 69 patients with IA and 30 healthy controls(HC)were recruited,among whom 30 had unruptured IA(UA)and 39 had ruptured IA(RA).First,plasma exosomes were extracted using a commercial exosome extraction kit,and the morphology and specific protein markers of exosomes were identified by transmission electron microscopy and Western blot analysis.Second,the miRNA expression profiles of plasma exosomes in 12 IA patients(4 UA and 8 RA)and 4 HC were determined using next-generation sequencing.In addition,significantly differentially expressed miRNAs were further analyzed by Quantitative Real-Time PCR(qRT-PCR)in a validation cohort of 99 subjects.Finally,the receiver operating characteristic(ROC)curves were used to evaluate the diagnostic potential of the miRNAs.Results In this study,isolated vesicle material was extracted in a shape having a substantially spherical shape with a diameter ranging from 100 to 150 nm;the vesicles expressed exosomal specific proteins CD9,HSC70 and TSG101.Second,from the sequencing analysis,181 miRNAs were identified to be differently(p < 0.05)expressed.Of these,9 miRNAs were up-regulated and 20 were down-regulated in patients with UA compared to HC.Also,21 were upregulated and 10 down-regulated in patients with RA compared to HC.In addition,compared with UA,92 miRNAs were up-regulated in RA whereas 29 were downregulated.Furthermore,qRT-PCR analysis confirmed that miR-145-5p and miR-29a-3p were upregulated in IA samples.To distinguish IA patients from controls,the area under the ROC curve(AUC)was 0.791 for miR-29a-3p,while that of miRNA-145-5p was 0.773 in terms of discriminating whether the aneurysm was ruptured.Multivariate logistic regression analysis showed that both miR-29a-5p and miR-145-5p were independent diagnostic indicators of intracranial aneurysms.Conclusions This study demonstrates for the first time that IA patients have specific plasma exosome miRNA expression profiles.Secondly,the selected miR-29a-3p and miR-145-5p have certain guiding significance for the diagnosis of IA and the prediction of IA rupture.