线粒体功能障碍已被证明在癌症生物学中发挥着关键作用。然而，其与肝内胆管癌（iCCA）的关系仍显着不足。30对iCCA和癌旁组织的RNA测序数据来自温州医科大学第一附属医院（WMU）。将 WMU 队列 (n = 30) 与公共 TCGA (n = 30) 和 GSE107943 (n = 30) 数据集整合，建立多中心 iCCA 队列。我们将 TCGA 和 GSE107943 队列合并为一个探索队列，以开发用于预后评估的线粒体特征，并利用 WMU 队列进行外部验证。基因本体论 (GO)、京都基因和基因组百科全书 (KEGG) 和 Hallmarker 分析用于 iCCA 相关线粒体相关基因 (MRG) 的功能解释。此外，还利用三个机构的数据进行无监督聚类，以识别基于线粒体的 iCCA 亚型。进一步研究线粒体功能障碍对药物反应、肿瘤免疫微环境改变和免疫反应的影响。 263 个 iCCA 相关 MRG 被鉴定与脂肪酸代谢、氧化磷酸化和免疫反应有关。细胞凋亡。通过单变量和多变量 Cox 以及 LASSO 分析，建立了具有 5 个最佳 MRG 的线粒体特征，用于评估探索队列中 AUC 值范围为 0.785 至 0.928 的 iCCA 患者的预后。该特征在 WMU 队列中也表现出令人满意的性能，AUC 值为 0.817-0.871，并被确定为两个队列中的独立风险预测因子。此外，我们发现线粒体评分较高且预后较差的患者的 CD4 T 细胞、NK 细胞和单核细胞浸润水平较低，并且对包括索拉非尼在内的靶向治疗表现出较高的敏感性。此外，还确定了两种基于线粒体的远距离亚型，其中亚型 2 与较短的生存时间和免疫抑制的肿瘤微环境有关。最后，通过免疫组织化学验证了五个关键 MRG 的差异蛋白表达。我们发现线粒体功能障碍调节 iCCA 中的异常代谢、氧化应激、免疫反应、细胞凋亡和药物敏感性。确定了线粒体特征和两种基于线粒体的 iCCA 亚型，用于临床风险分层和免疫表型分析。版权所有 © 2024 Chen、Lu、Chen、Zou、Bo、Li、Zhao、Zhao、Yu、Chen 和 Wu。

Mitochondrial dysfunction has been shown to play a critical role in cancer biology. However, its involvement in intrahepatic cholangiocarcinoma (iCCA) remains significantly understudied.RNA sequencing data of 30 pairs of iCCA and paracancerous tissues were collected from the First Affiliated Hospital of Wenzhou Medical University (WMU). The WMU cohort (n = 30) was integrated with public TCGA (n = 30) and GSE107943 (n = 30) datasets to establish a multi-center iCCA cohort. We merged the TCGA and GSE107943 cohorts into an exploration cohort to develop a mitochondria signature for prognosis assessment, and utilized the WMU cohort for external validation. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Hallmarker analyses were used for functional interpretation of iCCA associated mitochondria-related genes (MRGs). In addition, unsupervised clustering was performed to identify mitochondria-based iCCA subtypes with the data of three institutions. Further investigations were conducted to examine the impact of mitochondrial dysfunction on drug responses, alteration of the tumor immune microenvironment, and immune responses.Two hundred and sixty-three iCCA-related MRGs were identified to be related to fatty acid metabolism, oxidative phosphorylation, and apoptosis. Through univariate and multivariate Cox, and LASSO analyses, a mitochondria signature with five optimal MRGs was established to evaluate the prognosis of iCCA patients with the AUC values ranged from 0.785 to 0.928 in the exploration cohort. The signature also exhibited satisfactory performance in the WMU cohort with AUC values of 0.817-0.871, and was identified as an independent risk predictor in both cohorts. Additionally, we found that patients with higher mitochondria score with poor prognosis presented lower infiltration levels of CD4+ T-cell, NK cells, and monocytes, and demonstrated higher sensitivity to targeted therapies, including sorafenib. Furthermore, two distant mitochondria-based subtypes were determined, and subtype 2 was associated with shorter survival time and immunosuppressive tumor microenvironment. Finally, the differential protein expression of five key MRGs was verified by Immunohistochemistry.We found mitochondrial dysfunction modulates aberrant metabolism, oxidative stress, immune responses, apoptosis, and drug sensitivity in iCCA. A mitochondria signature and two mitochondria-based iCCA subtypes were identified for clinical risk stratification and immunophenotyping.Copyright © 2024 Chen, Lu, Chen, Zou, Bo, Li, Zhao, Zhao, Yu, Chen and Wu.