胆汁来源的小细胞外囊泡中的磷脂酰胆碱作为胆管癌的新型生物标志物。

Phosphatidylcholine in bile-derived small extracellular vesicles as a novel biomarker of cholangiocarcinoma.

机构信息

Second Department of Surgery, Hamamatsu University School of Medicine, Hamamatsu, Japan.

Department of Cellular and Molecular Anatomy, Hamamatsu University School of Medicine, Hamamatsu, Japan.

出版信息

Cancer Med. 2023 Jun;12(12):13007-13018. doi: 10.1002/cam4.5973. Epub 2023 Apr 25.

DOI:10.1002/cam4.5973

PMID:37096775

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10315768/

Abstract

BACKGROUND

Owing to the lack of definite diagnostic modalities, it is challenging to distinguish malignant cases of cholangiocarcinoma (CCA), which often causes biliary tract obstruction, from benign ones. Here, we investigated a novel lipid biomarker of CCA in bile-derived small extracellular vesicles (sEVs) and developed a simple detection method for clinical application.

METHODS

Bile samples from seven patients with malignant diseases (hilar CCA = 4, distal CCA = 3) and eight patients with benign diseases (gallstones = 6, primary sclerosing cholangitis = 1, autoimmune pancreatitis = 1) were collected through a nasal biliary drainage tube. sEVs were isolated via serial ultracentrifugation and characterized using nanoparticle tracking analysis, transmission electron microscopy, and immunoblotting (with CD9, CD63, CD81, and TSG101). Comprehensive lipidomic analysis was performed using liquid chromatography-tandem mass spectrometry. Using a measurement kit, we further confirmed whether lipid concentrations could be used as a potential CCA marker.

RESULTS

Lipidomic analysis of bile sEVs in the two groups identified 209 significantly increased lipid species in the malignant group. When focusing on lipid class, phosphatidylcholine (PC) level was 4.98-fold higher in the malignant group than in the benign group (P = 0.037). The receiver operating characteristic (ROC) curve showed a sensitivity of 71.4%, a specificity of 100%, and an area under the curve (AUC) of 0.857 (95% confidence interval [CI]:0.643-1.000). Using a PC assay kit, the ROC curve showed a cutoff value of 16.1 μg/mL, a sensitivity of 71.4%, a specificity of 100%, and an AUC of 0.839 (95% CI: 0.620-1.000).

CONCLUSION

PC level in sEVs from human bile is a potential diagnostic marker for CCA and can be assessed by a commercially available assay kit.

摘要

背景

由于缺乏明确的诊断方式，区分常导致胆道阻塞的胆管癌（CCA）的恶性病例与良性病例具有挑战性。在这里，我们研究了胆汁衍生的小细胞外囊泡（sEVs）中 CCA 的新型脂质生物标志物，并开发了一种简单的临床应用检测方法。

方法

通过鼻胆管引流管收集了 7 名恶性疾病患者（肝门部 CCA=4，远端 CCA=3）和 8 名良性疾病患者（胆结石=6，原发性硬化性胆管炎=1，自身免疫性胰腺炎=1）的胆汁样本。通过连续超速离心分离 sEVs，并通过纳米颗粒跟踪分析、透射电子显微镜和免疫印迹（使用 CD9、CD63、CD81 和 TSG101）进行表征。使用液相色谱-串联质谱进行全面脂质组学分析。使用试剂盒进一步确认脂质浓度是否可用作潜在的 CCA 标志物。

结果

两组胆汁 sEVs 的脂质组学分析确定了恶性组中 209 种显著增加的脂质种类。当关注脂质类别时，恶性组的磷脂酰胆碱（PC）水平比良性组高 4.98 倍（P=0.037）。接收器操作特征（ROC）曲线显示敏感性为 71.4%，特异性为 100%，曲线下面积（AUC）为 0.857（95%置信区间[CI]：0.643-1.000）。使用 PC 检测试剂盒，ROC 曲线显示截断值为 16.1μg/mL，敏感性为 71.4%，特异性为 100%，AUC 为 0.839（95%CI：0.620-1.000）。