MicroRNAs Derived From Circulating Exosomes As Non-invasive Biomarkers For Screening Three Kinds Of Pleural Effusions

Background: Pleural effusion is a common and complicate complication with poor prognosis,which secondly caused by pulmonary and pleural diseases.The etiology must be defined exactly to get proper therapies.In China,the tuberculous and malignant pleural effusions are frequently found in exudate.Nowadays,it's still difficult to reach an exact diagnosis after examination of X-line,CT-scan and pathologic diagnosis.Pleural biopsy and pleuroscopy are set as golden standard diagnosis of pleural effusion with high identification accuracy,but large range of trauma and high risk.Several soluble biomarkers attract much attention,such as interleukin,CEA and endothelin,it's still difficult to achieve satisfactory diagnosis with diverse sources,lack of traceability,not stable,low sensitivity and specificity.Exosomes extremely increase the stability of the mRNA and other bioactive molecule with the bonding of Ago2 in RISC and physical barrier,which give exosomes the superior properties of stabilization,high content,being traceable and noninvasive acting as diagnostic marker.We plan to collect exosomes from three kinds of pleural effusions(tuberculous,lung adenocarcinoma and benign pleural effusions),to perform the screening test which can be used to identify different pleural effusions,and to analyze the potential clinical value of the miRNA screening test.Objective: To get the characteristic miRNA screening test can be used to identify tuberculous,lung adenocarcinoma and benign pleural effusions.Methods: Three kinds of exosomes from pleural effusions were collected,and the exosomal miRNA were tested by deep sequencing of miRNA.Then characteristic miRNA spectrum were screened out and subsequently verified by quantitative RT-PCR.Results:(1)Extracts from pleural effusions were identified as exosomes.(2)Significantly differently expressed miRNAs among tuberculous,lung adenocarcinoma and benign pleural effusions were identified by deep sequencing of miRNA,which led to the selection of 12 miRNAs to perform a screening test.Compared with tuberculous and benign pleural effusions,miRNAs(miR-483-5p,miR-205-5p,miR-375,miR-200c-3p,miR-429,miR-200b-3p,miR-200a-3p,miR-203a-3p and miR-141-3p)from lung adenocarcinoma pleural effusions were presented in high levels,whereas miR-148a-3p,miR-451 a and miR-150-5p was most differently expressed between tuberculous and benign pleural effusions.(3)Results of verification test from quantitative RT-PCR arrived at almost the same expression pattern except miR-148a-3p and miR-205-5p.Conclusion: This analysis led to the selection of 12 miRNAs to perform a screening test for differential diagnoses of lung adenocarcinomas,pulmonary tuberculosis and benign pleural effusions.