Identification of Circulating MicroRNAs in Biliary Atresia by Next-Generation Sequencing.

Objectives: Biliary atresia (BA) is an idiopathic neonatal liver disease, characterized by inflammatory and fibrotic obliteration of extrahepatic bile ducts. Therefore, reliable methods for noninvasive diagnosis are needed. This study aimed to analyze circulating microRNAs (miRNAs) in patients with BA using next-generation sequencing (NGS) for identifying novel diagnostic biomarkers. Methods: An initial screening of miRNAs in plasma from patients with BA and healthy controls (HCs) was performed on an Illumina NGS platform. Differential miRNAs were validated by quantitative real-time PCR (qPCR). Target genes and related signal transduction pathways of differential miRNAs were predicted by online software. Results: In total, 146 differential miRNAs were identified by deep sequencing. Fifteen miRNAs with read counts more than 1000, that included 7 upregulated and 8 downregulated miRNAs, were predicted to be associated with liver fibrosis, biliary differentiation and bile duct development. Of these, six miRNAs with read counts more than 5000 were analyzed by qPCR on an independent sample set comprising 44 patients with BA, 20 cholestatic disease controls and 20 HCs. Two up-regulated miRNAs (miR-122-5p, miR-100-5p) and two down-regulated miRNAs (miR-140-3p, miR-126-3p) were confirmed by individual qPCR. Only miR-140-3p was significantly different from controls (P

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