Huang Pinzhu, Wei Guangyan, Kirkpatrick Jesse D, Lin Yi, Tan Li, Matta Heansika, Nasser Imad, Huang Mingzhe, Chen Li, Petitjean Mathieu, Skelton-Badlani Disha, Gao Wen, Vaid Kahini, Zhao Shuangshuang, Lugovskoy Alicia, Alenzi Maram, Chen Xin, Gores Gregory J, Popov Yury V
Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Department of Colon and Rectum Surgery, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Division of Gastroenterology and Hepatology, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Department of Radiation Oncology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
J Hepatol. 2025 Jan;82(1):84-96. doi: 10.1016/j.jhep.2024.07.016. Epub 2024 Jul 30.
BACKGROUND & AIMS: Cholangiocarcinoma (CCA) is a dreaded complication of primary sclerosing cholangitis (PSC) that is difficult to diagnose and associated with high mortality. A lack of animal models of CCA recapitulating the hepatic microenvironment of sclerosing cholangitis has hindered the development of novel treatments. Herein, we sought to develop a mouse model of PSC-associated CCA.
Ten-week-old Mdr2 mice with congenital PSC-like disease, and healthy wild-type littermates were subjected to either modified retrograde biliary instillation or hydrodynamic tail vein injection of a sleeping beauty transposon-transposase plasmid system with activated AKT (myr-AKT) and Yap (YapS127A) proto-oncogenes (SB AKT/YAP1). The role of TGFβ was interrogated via ALK5 inhibitor (SB-525334) administration. Tumor phenotype, burden and desmoplastic reaction were analyzed histologically and via RNA sequencing.
While SB AKT/YAP1 plasmids administered via retrograde biliary injection caused tumors in Mdr2, only 26.67% (4/15) of these tumors were CCA. Alternatively, hydrodynamic tail vein injection of SB AKT/YAP1 resulted in robust tumorigenesis in all fibrotic Mdr2 mice with high CCA burden compared to healthy mice. Tumors phenotypically resembled human CCA, expressed multiple CCA (but not hepatocellular carcinoma) markers, and exhibited a profound desmoplastic reaction. RNA sequencing analysis revealed profound transcriptional changes in CCA evolving in a PSC-like context, with specific alterations in multiple immune pathways. Pharmacological TGFβ inhibition led to enhanced immune cell tumor infiltration, reduced tumor burden and suppressed desmoplastic collagen accumulation compared to placebo.
We established a new high-fidelity cholangiocarcinoma model in mice, termed SB CCA.Mdr2, which recapitulates the increased susceptibility to CCA in the setting of biliary injury and fibrosis observed in PSC. Through transcriptomics and pharmacological studies, we show dysregulation of multiple immune pathways and TGFβ signaling as potential drivers of CCA in a PSC-like microenvironment.
Animal models for primary sclerosing cholangitis (PSC)-related cholangiocarcinoma (PSC-CCA) are lacking. Thus, we have developed and characterized a new mouse model of PSC-CCA, termed SB CCA.Mdr2, which features reliable tumor induction on a PSC-like background of biliary injury and fibrosis. Global gene expression alterations were identified and standardized tools, including automated whole slide image analysis methodology for tumor burden and feature analysis, were established to enable systematic research into PSC-CCA biology and formal preclinical drug testing.
胆管癌（CCA）是原发性硬化性胆管炎（PSC）的一种可怕并发症，难以诊断且死亡率高。缺乏能够重现硬化性胆管炎肝脏微环境的CCA动物模型阻碍了新型治疗方法的开发。在此，我们试图建立一种PSC相关CCA的小鼠模型。
将患有先天性PSC样疾病的10周龄Mdr2小鼠和健康的野生型同窝小鼠，通过改良逆行胆管灌注或尾静脉高压注射携带激活的AKT（myr-AKT）和Yap（YapS127A）原癌基因的睡美人转座子-转座酶质粒系统（SB AKT/YAP1）。通过给予ALK5抑制剂（SB-525334）来探究TGFβ的作用。通过组织学和RNA测序分析肿瘤表型、负荷和促结缔组织增生反应。
虽然通过逆行胆管注射给予的SB AKT/YAP1质粒在Mdr2小鼠中引发了肿瘤，但这些肿瘤中只有26.67%（4/15）是CCA。相比之下，与健康小鼠相比，尾静脉高压注射SB AKT/YAP1在所有具有高CCA负荷的纤维化Mdr2小鼠中都导致了强烈的肿瘤发生。肿瘤在表型上类似于人类CCA，表达多种CCA（而非肝细胞癌）标志物，并表现出明显的促结缔组织增生反应。RNA测序分析揭示了在类似PSC的背景下发展的CCA中存在深刻的转录变化，多个免疫途径有特定改变。与安慰剂相比，药物性TGFβ抑制导致免疫细胞肿瘤浸润增加、肿瘤负荷降低以及促结缔组织增生性胶原积累受到抑制。
我们在小鼠中建立了一种新的确保真度高的胆管癌模型，称为SB CCA.Mdr2，它重现了在PSC中观察到的胆管损伤和纤维化情况下对CCA易感性增加的情况。通过转录组学和药理学研究，我们表明多种免疫途径和TGFβ信号失调是类似PSC微环境中CCA的潜在驱动因素。
缺乏原发性硬化性胆管炎（PSC）相关胆管癌（PSC-CCA）的动物模型。因此，我们开发并鉴定了一种新的PSC-CCA小鼠模型，称为SB CCA.Mdr2，其特点是在类似PSC的胆管损伤和纤维化背景下能可靠地诱导肿瘤。识别了全局基因表达改变，并建立了标准化工具，包括用于肿瘤负荷和特征分析的自动全玻片图像分析方法，以促进对PSC-CCA生物学的系统研究和正式的临床前药物测试。
