Dwyer Benjamin J, Jarman Edward J, Gogoi-Tiwari Jully, Ferreira-Gonzalez Sofia, Boulter Luke, Guest Rachel V, Kendall Timothy J, Kurian Dominic, Kilpatrick Alastair M, Robson Andrew J, O'Duibhir Eoghan, Man Tak Yung, Campana Lara, Starkey Lewis Philip J, Wigmore Stephen J, Olynyk John K, Ramm Grant A, Tirnitz-Parker Janina E E, Forbes Stuart J
Centre for Regenerative Medicine, Scottish Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, UK; School of Pharmacy and Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, Bentley, WA, Australia.
MRC Human Genetics Unit, Western General Hospital Campus, Edinburgh, UK.
J Hepatol. 2021 Apr;74(4):860-872. doi: 10.1016/j.jhep.2020.11.018. Epub 2020 Nov 19.
BACKGROUND & AIMS: Cholangiocarcinoma (CCA) is a cancer of the hepatic bile ducts that is rarely resectable and is associated with poor prognosis. Tumour necrosis factor-like weak inducer of apoptosis (TWEAK) is known to signal via its receptor fibroblast growth factor-inducible 14 (Fn14) and induce cholangiocyte and myofibroblast proliferation in liver injury. We aimed to characterise its role in CCA.
The expression of the TWEAK ligand and Fn14 receptor was assessed immunohistochemically and by bulk RNA and single cell transcriptomics of human liver tissue. Spatiotemporal dynamics of pathway regulation were comprehensively analysed in rat and mouse models of thioacetamide (TAA)-mediated CCA. Flow cytometry, qPCR and proteomic analyses of CCA cell lines and conditioned medium experiments with primary macrophages were performed to evaluate the downstream functions of TWEAK/Fn14. In vivo pathway manipulation was assessed via TWEAK overexpression in NICD/AKT-induced CCA or genetic Fn14 knockout during TAA-mediated carcinogenesis.
Our data reveal TWEAK and Fn14 overexpression in multiple human CCA cohorts, and Fn14 upregulation in early TAA-induced carcinogenesis. TWEAK regulated the secretion of factors from CC-SW-1 and SNU-1079 CCA cells, inducing polarisation of proinflammatory CD206 macrophages. Pharmacological blocking of the TWEAK downstream target chemokine monocyte chemoattractant protein 1 (MCP-1 or CCL2) significantly reduced CCA xenograft growth, while TWEAK overexpression drove cancer-associated fibroblast proliferation and collagen deposition in the tumour niche. Genetic Fn14 ablation significantly reduced inflammatory, fibrogenic and ductular responses during carcinogenic TAA-mediated injury.
These novel data provide evidence for the action of TWEAK/Fn14 on macrophage recruitment and phenotype, and cancer-associated fibroblast proliferation in CCA. Targeting TWEAK/Fn14 and its downstream signals may provide a means to inhibit CCA niche development and tumour growth.
Cholangiocarcinoma is an aggressive, chemotherapy-resistant liver cancer. Interactions between tumour cells and cells that form a supportive environment for the tumour to grow are a source of this aggressiveness and resistance to chemotherapy. Herein, we describe interactions between tumour cells and their supportive environment via a chemical messenger, TWEAK and its receptor Fn14. TWEAK/Fn14 alters the recruitment and type of immune cells in tumours, increases the growth of cancer-associated fibroblasts in the tumour environment, and is a potential target to reduce tumour formation.
胆管癌（CCA）是一种肝内胆管癌，很少能切除，预后较差。已知肿瘤坏死因子样凋亡微弱诱导剂（TWEAK）通过其受体成纤维细胞生长因子诱导14（Fn14）发出信号，并在肝损伤中诱导胆管细胞和成肌纤维细胞增殖。我们旨在阐明其在CCA中的作用。
通过免疫组织化学以及对人肝组织进行批量RNA和单细胞转录组学分析，评估TWEAK配体和Fn14受体的表达。在硫代乙酰胺（TAA）介导的CCA大鼠和小鼠模型中，全面分析通路调节的时空动态。对CCA细胞系进行流式细胞术、qPCR和蛋白质组学分析，并对原代巨噬细胞进行条件培养基实验，以评估TWEAK/Fn14的下游功能。通过在NICD/AKT诱导的CCA中过表达TWEAK或在TAA介导的致癌过程中进行Fn14基因敲除，评估体内通路操作。
我们的数据显示，在多个人类CCA队列中TWEAK和Fn14过表达，且在早期TAA诱导的致癌过程中Fn14上调。TWEAK调节CC-SW-1和SNU-1079 CCA细胞因子的分泌，诱导促炎性CD206巨噬细胞极化。对TWEAK下游靶标趋化因子单核细胞趋化蛋白1（MCP-1或CCL2）进行药理阻断可显著降低CCA异种移植瘤的生长，而TWEAK过表达则促使肿瘤微环境中癌症相关成纤维细胞增殖和胶原沉积。在致癌性TAA介导的损伤过程中，基因敲除Fn14可显著降低炎症、纤维化和小胆管反应。
这些新数据为TWEAK/Fn14在CCA中对巨噬细胞募集和表型以及癌症相关成纤维细胞增殖的作用提供了证据。靶向TWEAK/Fn14及其下游信号可能提供一种抑制CCA微环境发育和肿瘤生长的方法。
胆管癌是一种侵袭性、化疗耐药的肝癌。肿瘤细胞与为肿瘤生长形成支持性环境的细胞之间的相互作用是这种侵袭性和化疗耐药性的来源。在此，我们描述了肿瘤细胞与其支持性环境之间通过化学信使TWEAK及其受体Fn14的相互作用。TWEAK/Fn14改变肿瘤中免疫细胞的募集和类型，增加肿瘤环境中癌症相关成纤维细胞的生长，是减少肿瘤形成的潜在靶点。
