Han Jin-Yi, Ahn Keun Soo, Kim Yong Hoon, Kim Tae-Seok, Baek Won-Ki, Suh Seong-Il, Kang Koo Jeong
Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, Keimyung University Dongsan Medical Center, Daegu, Korea.
Department of Microbiology, Keimyung University School of Medicine, Daegu, Korea.
Ann Surg Treat Res. 2021 Sep;101(3):140-150. doi: 10.4174/astr.2021.101.3.140. Epub 2021 Aug 31.
In this pilot study, using next-generation sequencing and integrated messenger RNA (mRNA) sequencing, we investigated circulating microRNA (miRNA) expression profiling from bile-derived exosomes to identify dysregulated miRNA signatures and oncogenic pathways and determine their effects on targeted mRNAs in cholangiocarcinoma (CCA). Moreover, we explored the possibility that genetic analysis using bile-derived exosomes may replace gene analysis using tissue.
Bile was collected from a patient with perihilar CCA before curative resection. As a control, bile was collected from a patient with a common bile duct stone. Exosomes were isolated from the bile, and we performed next-generation miRNA sequencing using isolated exosomes. To evaluate miRNA-mRNA interactions, mRNA sequencing was performed using bile fluid in both patients.
We identified 22 differentially expressed miRNAs. More than 65% of the predicted mRNA targets of those miRNAs were actually differentially expressed between control and CCA bile samples. In functional pathway analysis, targets of 22 miRNAs were primarily enriched in mitogen-activated protein kinase, platelet derived growth factor, vascular endothelial growth factor, epidermal growth factor receptor, and p53 signaling. In particular, in the functional assessment of miRNA-mRNA interactions, RAS pathways, including downstream pathways (PI3K-AKT-mTOR and RAS-RAF-MEK-ERK), were determined to be enriched.
Circulating miRNAs in bile-derived exosomes provide new information for the development of miRNA analysis in CCA. These miRNAs may represent the oncogenic characteristics of CCA tissue, enabling them to be used instead of tissue samples for the diagnosis of CCA. Further research investigating circulating miRNAs in bile exosomes may lead to more rational, targeted approaches to treatment.
在这项初步研究中，我们使用下一代测序和整合信使核糖核酸（mRNA）测序技术，研究了胆汁来源外泌体中循环微核糖核酸（miRNA）的表达谱，以识别失调的miRNA特征和致癌途径，并确定它们对胆管癌（CCA）中靶向mRNA的影响。此外，我们探讨了使用胆汁来源外泌体进行基因分析可能替代使用组织进行基因分析的可能性。
在根治性切除术前，从一名肝门部CCA患者收集胆汁。作为对照，从一名胆总管结石患者收集胆汁。从胆汁中分离出外泌体，并使用分离出的外泌体进行下一代miRNA测序。为了评估miRNA与mRNA的相互作用，对两名患者的胆汁进行了mRNA测序。
我们鉴定出22种差异表达的miRNA。这些miRNA预测的mRNA靶标中，超过65%在对照和CCA胆汁样本之间实际存在差异表达。在功能通路分析中，22种miRNA的靶标主要富集在丝裂原活化蛋白激酶、血小板衍生生长因子、血管内皮生长因子、表皮生长因子受体和p53信号通路中。特别是，在miRNA与mRNA相互作用的功能评估中，确定RAS通路（包括下游通路PI3K-AKT-mTOR和RAS-RAF-MEK-ERK）富集。
胆汁来源外泌体中的循环miRNA为CCA中miRNA分析的发展提供了新信息。这些miRNA可能代表CCA组织的致癌特征，使其能够替代组织样本用于CCA的诊断。进一步研究胆汁外泌体中的循环miRNA可能会带来更合理、有针对性的治疗方法。
