■线粒体功能障碍已被证明在癌症生物学中起关键作用。然而,其在肝内胆管癌(iCCA)中的参与仍未得到充分研究。
■30对iCCA和癌旁组织的RNA测序数据来自温州医科大学附属第一医院(WMU)。将WMU队列(n=30)与公共TCGA(n=30)和GSE107943(n=30)数据集整合以建立多中心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(MRGs)的功能解释。此外,使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。进行了进一步的研究,以检查线粒体功能障碍对药物反应的影响,肿瘤免疫微环境的改变,和免疫反应。
■确定了二百六十三个与iCCA相关的MRG与脂肪酸代谢有关,氧化磷酸化,和凋亡。通过单变量和多变量Cox,和LASSO分析,在探索队列中,我们建立了5个最佳MRGs的线粒体特征来评估iCCA患者的预后,这些患者的AUC值在0.785~0.928之间.该特征在AUC值为0.817-0.871的WMU队列中也表现出令人满意的表现,并在两个队列中被确定为独立的风险预测因子。此外,我们发现线粒体评分较高、预后较差的患者CD4+T细胞浸润水平较低,NK细胞,和单核细胞,并对靶向治疗表现出更高的敏感性,包括索拉非尼.此外,确定了两个远处的线粒体亚型,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,5个关键MRGs的差异蛋白表达通过免疫组织化学证实。
■我们发现线粒体功能障碍调节异常代谢,氧化应激,免疫反应,凋亡,和iCCA中的药物敏感性。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。
UNASSIGNED: Mitochondrial dysfunction has been shown to play a critical role in cancer biology. However, its involvement in intrahepatic cholangiocarcinoma (iCCA) remains significantly understudied.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.