Detection of cholangiocarcinoma (CCA) remains a diagnostic challenge. We established diagnostic peptide biomarkers in bile and urine based on capillary electrophoresis coupled to mass spectrometry (CE-MS) to detect both local and systemic changes during CCA progression. In a prospective cohort study we recently demonstrated that combined bile and urine proteome analysis could further improve diagnostic accuracy of CCA diagnosis in patients with unknown biliary strictures. As a continuation of these investigations, the aim of the present study was to investigate the pathophysiological mechanisms behind the molecular determinants reflected by bile and urine peptide biomarkers. Protease mapping and gene ontology cluster analysis were performed for the previously defined CE-MS based biomarkers in bile and urine. For that purpose, bile and urine peptide profiles (from samples both collected at the date of endoscopy) were investigated from a representative cohort of patients with benign (n = 76) or CCA-associated (n = 52) biliary strictures (verified during clinical follow-up). This was supplemented with a literature search for the association of the individual biomarkers included in the proteomic patterns with CCA or cancer progression. For most of the peptide markers, association to CCA has been described in literature. Protease mapping revealed ADAMTS4 activity in cleavage of both bile and urine CCA peptide biomarkers. Furthermore, increased chymase activity in bile points to mast cell activation at the tumor site. Gene ontology cluster analysis indicates cellular response to chemical stimuli and stress response as local and extracellular matrix reorganization by tissue destruction and repair as systemic events. The analysis further supports that the mapped proteases are drivers of local and systemic events. The study supports connection of the CCA-associated peptide biomarkers to the molecular pathophysiology and indicates an involvement in epithelial-to-mesenchymal transition, generation of cancer-associated fibroblasts and activation of residual immune cells. Proteases, extracellular matrix components, inflammatory cytokines, proangiogenic, growth and vasoactive factors released from the tumor microenvironment are drivers of systemic early events during CCA progression.

中文翻译：

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胆汁和尿液肽标记物谱：胆管癌分子通路和生物学过程的访问键

胆管癌（CCA）的检测仍然是诊断上的挑战。我们基于毛细管电泳结合质谱（CE-MS）在胆汁和尿液中建立了诊断性肽生物标志物，以检测CCA进展过程中的局部和全身性变化。在一项前瞻性队列研究中，我们最近证明了结合胆汁和尿液蛋白质组分析可以进一步提高胆道狭窄未知患者的CCA诊断的诊断准确性。作为这些研究的延续，本研究的目的是研究胆汁和尿肽生物标志物反映的分子决定因素背后的病理生理机制。对胆汁和尿液中先前定义的基于CE-MS的生物标记物进行了蛋白酶作图和基因本体聚类分析。为了这个目的，胆汁和尿液肽谱（均来自内窥镜检查时收集的样品）是从具有代表性的良性（n = 76）或CCA相关（n = 52）胆管狭窄（在临床随访中验证）患者的队列研究。补充文献搜索以寻找蛋白质组学模式中包括的单个生物标志物与CCA或癌症进展的关联。对于大多数肽标记物，在文献中已经描述了与CCA的缔合。蛋白酶作图揭示了ADAMTS4在胆汁和尿液CCA肽生物标志物裂解中的活性。此外，胆汁中的糜蛋白酶活性增加表明肿瘤部位的肥大细胞活化。基因本体簇分析表明细胞对化学刺激的反应和应激反应是通过组织破坏和修复作为系统性事件而进行的局部和细胞外基质重组。该分析进一步支持所定位的蛋白酶是局部和全身事件的驱动器。该研究支持CCA相关肽生物标志物与分子病理生理学的联系，并表明参与上皮到间充质转化，癌症相关成纤维细胞的产生和残余免疫细胞的活化。从肿瘤微环境释放的蛋白酶，细胞外基质成分，炎性细胞因子，促血管生成，生长和血管活性因子是CCA进展过程中系统性早期事件的驱动因素。该分析进一步支持所定位的蛋白酶是局部和全身事件的驱动器。该研究支持CCA相关肽生物标志物与分子病理生理学的联系，并表明参与上皮到间充质转化，癌症相关成纤维细胞的产生和残余免疫细胞的活化。从肿瘤微环境释放的蛋白酶，细胞外基质成分，炎性细胞因子，促血管生成，生长和血管活性因子是CCA进展过程中系统性早期事件的驱动因素。该分析进一步支持所定位的蛋白酶是局部和全身事件的驱动器。该研究支持CCA相关肽生物标志物与分子病理生理学的联系，并表明参与上皮到间充质转化，癌症相关成纤维细胞的产生和残余免疫细胞的活化。从肿瘤微环境释放的蛋白酶，细胞外基质成分，炎性细胞因子，促血管生成，生长和血管活性因子是CCA进展过程中系统性早期事件的驱动因素。癌症相关的成纤维细胞的产生和残余免疫细胞的活化。从肿瘤微环境释放的蛋白酶，细胞外基质成分，炎性细胞因子，促血管生成，生长和血管活性因子是CCA进展过程中系统性早期事件的驱动因素。癌症相关的成纤维细胞的产生和残余免疫细胞的活化。从肿瘤微环境释放的蛋白酶，细胞外基质成分，炎性细胞因子，促血管生成，生长和血管活性因子是CCA进展过程中系统性早期事件的驱动因素。