Circulating miRNA is a useful diagnostic biomarker for nonalcoholic steatohepatitis in nonalcoholic fatty liver disease

Tae Hyung Kim,Baek-Hui Kim,Yoonseok Lee,Soon Ho Um,Moon Young Kim,Hyung Joon Yim,Jong Eun Yeon,Yeon Seok Seo,Hayeon Kim,Sun Young Yim,Eunjung Ko,Young Kul Jung,Kwan Soo Byun,Young-Sun Lee,Seonghee Kang,Ji Hoon Kim,Jeong-An Gim

doi:10.1038/s41598-021-94115-6

Abstract

Nonalcoholic steatohepatitis (NASH) is considered as a progressive form of nonalcoholic fatty liver disease (NAFLD). To distinguish NASH from nonalcoholic fatty liver (NAFL), we evaluated the diagnostic value of circulating miRNAs. Small RNA sequencing was performed on 12 NAFL patients and 12 NASH patients, and the miRNA expression was compared. After selecting miRNAs for the diagnosis of NASH, we analyzed the diagnostic accuracy of each miRNA and the combination of miRNAs. External validation was performed using quantitative reverse transcription PCR. Among the 2,588 miRNAs, 26 miRNAs significantly increased in the NASH group than in the NAFL group. Among the 26 elevated miRNAs in the NASH group, 8 miRNAs were selected, and in silico analysis was performed. Only four miRNAs (miR-21-5p, miR-151a-3p, miR-192-5p, and miR-4449) showed significant area under the receiver operating characteristic curve (AUC) values for NASH diagnosis. The combination of the four miRNAs showed satisfactory diagnostic accuracy for NASH (AUC 0.875; 95% CI 0.676–0.973). External validation revealed similar diagnostic accuracy for NASH (AUC 0.874; 95% CI 0.724–0.960). NASH represents significantly distinct miRNA expression profile compared with NAFL. The combination of serum circulating miRNAs can be used as a novel biomarker for the NASH diagnosis in NAFLD.

Highlights

Nonalcoholic steatohepatitis (NASH) is considered as a progressive form of nonalcoholic fatty liver disease (NAFLD)
For miRNA sequencing, sera from 24 biopsy-proven patients with NAFLD were collected between February 2019 and July 2019, including 12 patients with NAFL and 12 patients with NASH
Ballooning is essential for the diagnosis of NASH; there was a significant difference in the presence of ballooning between the NAFL and NASH groups

Summary

Introduction

Nonalcoholic steatohepatitis (NASH) is considered as a progressive form of nonalcoholic fatty liver disease (NAFLD). To distinguish NASH from nonalcoholic fatty liver (NAFL), we evaluated the diagnostic value of circulating miRNAs. Small RNA sequencing was performed on 12 NAFL patients and 12 NASH patients, and the miRNA expression was compared. The combination of serum circulating miRNAs can be used as a novel biomarker for the NASH diagnosis in NAFLD. Nonalcoholic steatohepatitis (NASH) is considered a progressive disease of NAFLD, which is distinguished from non-alcoholic fatty liver (NAFL)[3]. Diagnostic accuracy to distinguish NASH from NAFL was evaluated using miR-34a, but it showed moderate accuracy [area under the receiver operating characteristic curve (AUC) = 0.78)]. We aimed to develop a biomarker to diagnose NASH in NAFLD by analyzing the circulating miRNA expression profile in sera from patients with biopsy-confirmed NAFLD using emerging generation sequencing (NGS)

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