The Department of Infectious Diseases, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Front Cell Infect Microbiol. 2022 Jul 7;12:923300. doi: 10.3389/fcimb.2022.923300. eCollection 2022.
The small noncoding RNAs (sncRNAs) including microRNAs and the noncanonical sncRNAs [i.e., tRNA-derived small RNAs (tsRNAs) and rRNA-derived small RNAs (rsRNAs)] are a vital class of gene regulators in response to a variety of diseases. We focus on an sncRNA signature enriched in hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF) to develop a plasma exosome-based noninvasive biomarker for human ACLF.
In this work, sncRNAs related to HBV-ACLF were identified by small RNA sequencing (RNA-seq) in plasma exosomes collected from 3 normal subjects, 4 chronic hepatitis B (CHB) patients with flare, and 6 HBV-ACLF patients in the discovery cohort. Thereafter, the differentially expressed sncRNAs were further verified in a validation cohort (n = 313) using the newly developed molecular signature incorporating different mi/ts/rsRNAs (named as MTR-RNAs) through qRT-PCR assays. Subsequently, using the least absolute shrinkage and selection operator (LASSO) logistic regression (LR) model analysis, we developed an MTR-RNA classifier for early detection of ACLF.
The identified sncRNAs (hsa-miR-23b-3p, hsa-miR-223-3p, hsa-miR-339-5p, tsRNA-20, tsRNA-46, and rsRNA-249) were specifically differentially expressed in plasma exosomes of HBV-ACLF. The MTR-RNA signature (AUC = 0.787) containing the above sncRNAs distinguished HBV-ACLF cases among normal subjects with 71.67% specificity and 74.29% sensitivity, CHB patients with flare (AUC = 0.694, 85.71% sensitivity/59.5% specificity), and patients with CHB/cirrhosis (AUC = 0.785, 57.14% sensitivity/94.59% specificity). Notably, it revealed 100% specificity/94.80% sensitivity in detecting patients or normal people.
Our as-constructed plasma-derived exosomal sncRNA signature can serve as a reliable biomarker for ACLF detection and also be adopted to be the pre-triage biomarker for selecting cases that can gain benefits from adjuvant treatment.
小非编码 RNA（sncRNA）包括 microRNA 和非规范 sncRNA[即 tRNA 衍生的小 RNA（tsRNA）和 rRNA 衍生的小 RNA（rsRNA）]是一类重要的基因调节剂，可响应多种疾病。我们专注于富含乙型肝炎病毒（HBV）相关慢加急性肝衰竭（ACLF）的 sncRNA 特征，以开发一种基于血浆外泌体的人类 ACLF 无创生物标志物。
在本研究中，通过对来自 3 名正常对照、4 名慢性乙型肝炎（CHB）患者爆发和 6 名 HBV-ACLF 患者的血浆外泌体进行小 RNA 测序（RNA-seq），鉴定出与 HBV-ACLF 相关的 sncRNA。此后，使用新开发的分子特征（命名为 MTR-RNAs），通过 qRT-PCR 检测，在验证队列（n=313）中进一步验证差异表达的 sncRNA。随后，使用最小绝对收缩和选择算子（LASSO）逻辑回归（LR）模型分析，我们开发了一种用于早期检测 ACLF 的 MTR-RNA 分类器。
在 HBV-ACLF 患者的血浆外泌体中，特异性差异表达的 sncRNA（hsa-miR-23b-3p、hsa-miR-223-3p、hsa-miR-339-5p、tsRNA-20、tsRNA-46 和 rsRNA-249）。包含上述 sncRNA 的 MTR-RNA 特征（AUC=0.787）能够区分 HBV-ACLF 患者与正常对照（特异性为 71.67%，敏感性为 74.29%）、CHB 患者爆发（AUC=0.694，敏感性为 85.71%，特异性为 59.5%）和 CHB/肝硬化患者（AUC=0.785，敏感性为 57.14%，特异性为 94.59%）。值得注意的是，该特征在检测患者或正常人时具有 100%的特异性/94.80%的敏感性。
我们构建的血浆源性外泌体 sncRNA 特征可作为 ACLF 检测的可靠生物标志物，并可作为辅助治疗获益病例的预分诊生物标志物。
