A new mouse model for studying the pathogenesis and immunobiology of intrahepatic cholangiocarcinoma and improving its immunotherapy

研究肝内胆管癌发病机制和免疫生物学并改进其免疫治疗的新小鼠模型

• 批准号: 10711615
• 负责人: Michael Karin
• 金额: $ 56.39万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711615
• 关键词: Accounting; Alcohol abuse; Biliary; CD8-Positive T-Lymphocytes; Cell Communication; Cell Nucleus; Cell Surface Receptors; Cells; Characteristics; Cholestasis; Clinical; Clinical Management; Complement; Complication; Data; Desmoplastic; Diagnosis; Differential Diagnosis; Early Diagnosis; Epigenetic Process; Evaluation; Evolution; Exclusion; Exhibits; Failure; Fibroblasts; Goals; Hepatobiliary; Hepatocyte; High Fat Diet; Human; Immune; Immune checkpoint inhibitor; Immunobiology; Immunocompetent; Immunophenotyping; Immunosuppression; Immunotherapy; Incidence; Inflammatory Bowel Diseases; Intraepithelial Neoplasia; Intrahepatic Cholangiocarcinoma; KDR gene; Lesion; Malignant - descriptor; Malignant Neoplasms; Maps; Mediating; Modeling; Molecular; Mus; Mutation; Myeloid-derived suppressor cells; Nodule; Non-Insulin-Dependent Diabetes Mellitus; Oncogenic; Pathogenesis; Pathologic; Primary Malignant Neoplasm of Liver; Primary carcinoma of the liver cells; Radiology Specialty; Reaction; Risk Factors; Signal Transduction; Study models; Survival Rate; T cell receptor repertoire sequencing; T-Lymphocyte; T-cell receptor repertoire; Techniques; Therapeutic; Transcriptional Activation; Translating; Tumor-Infiltrating Lymphocytes; Variant; Vegf Inhibitor; biliary tract; cancer cell; cancer type; checkpoint inhibition; cholangiocyte; endoplasmic reticulum stress; experimental study; immunogenic; improved; information gathering; inhibitor; mouse model; neoplastic; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; overexpression; pressure; prevent; progenitor; regenerative; response; tobacco abuse; transcription factor; transcriptomics; tumor; tumor microenvironment; tumor-immune system interactions

ABSTRACT Intrahepatic cholangiocarcinoma (ICC) is an aggressive primary liver cancer (PLC) that is particularly hard to treat, having an overall 5-year survival rate of 8%. Like the major PLC, hepatocellular carcinoma (HCC), ICC incidence is on the rise due to the ongoing increase in cholestatic liver diseases, non-alcoholic fatty liver diseases, type 2 diabetes and alcohol and tobacco abuse. Unlike HCC, ICC is often detected at an advanced stage when survival prospects are poor, a complication of its imprecise differentiation from early HCC by current radiological techniques. Early differential diagnosis of two PLCs is critical due to their distinct management and the absence of targetable oncogenic drivers shared by ICC and HCC. This problem could be solved by identification of a PLC- type agnostic therapeutic approach, such as immune checkpoint inhibition (ICI), that is applicable to both ICC and HCC. However, while ICI based therapies were approved for HCC and their efficacy has been improved by combination with VEGF inhibitors, they have performed poorly in ICC. This failure can be attributed, in part, to poor mechanistic understanding of the ICC-specific immunosuppressive tumor microenvironment (TME) and lack of suitable mouse models that allow the evaluation of ICI in combination with therapeutics capable of dismantling immunosuppression. We have solved this problem by developing a new ICC model that unlike previous mouse models does not depend on forced overexpression of potent oncogenic drivers that rapidly induce ICC in the absence of selective pressure for acquisition of additional mutations. Our MUP-uPA/NRF2Act model depends on combination of ER stress with activation of transcription factor NRF2 in bipotential hepatobiliary progenitors and mature hepatocytes and shows robust, highly penetrant, human-like stepwise progression towards ICC that is accompanied by the buildup of an immunosuppressive TME exhibiting CD8+ T cell exclusion. We plan to establish the MUP-uPA/NRF2Act mouse as the leading model for studying the evolution of ICC and its immunosuppressive TME and for finding treatments that will dismantle immunosuppression and boost the response to existing PD-(L)1 inhibitors. To accomplish this goal, we will fully characterize the MUP- uPA/NRF2Act mouse, defining the transcriptomic, epigenetic, and genetic alterations at each stage of ICC progression, including ductular reactions, biliary intraepithelial neoplasia and established desmoplastic cancer. We will also define the T cell receptor (TCR) repertoire of tumor infiltrating lymphocytes at each stage of malignant progression and mine the transcriptomic data for cancer cell produced factors that mediate desmoplasia, immunosuppression and CD8+ T cell exclusion. These studies will be complemented by immunophenotyping and immunodepletion experiments whose goal is the identification of targetable molecular switches responsible for establishment of the ICC-specific immunosuppressive TME. We will use this information to identify clinically approved treatments that can overcome immunosuppression and vastly improve the efficacy of ICI therapy based on PD-(L)1 inhibitors.

抽象的 肝内胆管癌 (ICC) 是一种侵袭性原发性肝癌 (PLC)，特别难以治疗。 治疗，总体 5 年生存率为 8%。像主要的PLC、肝细胞癌（HCC）、ICC 由于胆汁淤积性肝病、非酒精性脂肪肝病的持续增加，发病率呈上升趋势， 2 型糖尿病以及酗酒和吸烟。与 HCC 不同，ICC 通常在晚期才被发现 生存前景不佳，这是目前放射学无法准确区分早期 HCC 的并发症 技术。两个 PLC 的早期鉴别诊断至关重要，因为它们的管理不同且缺乏 ICC 和 HCC 共有的目标致癌驱动因素。这个问题可以通过识别PLC来解决 类型不可知的治疗方法，例如免疫检查点抑制 (ICI)，适用于 ICC 和肝癌。然而，虽然基于 ICI 的疗法已被批准用于 HCC，并且其疗效已通过以下方法得到改善： 与 VEGF 抑制剂联合使用时，它们在 ICC 中表现不佳。这种失败的部分原因可归咎于 对 ICC 特异性免疫抑制肿瘤微环境 (TME) 的机制了解不足， 缺乏合适的小鼠模型来评估 ICI 与能够联合治疗的药物 解除免疫抑制。我们通过开发一种新的 ICC 模型解决了这个问题，该模型不同于 以前的小鼠模型并不依赖于强效致癌驱动因素的强制过度表达，这些驱动因素会迅速 在没有选择压力的情况下诱导 ICC 以获得额外的突变。我们的 MUP-uPA/NRF2 法案 模型取决于 ER 应激与双电位转录因子 NRF2 激活的组合 肝胆祖细胞和成熟肝细胞，显示出稳健、高渗透性、类似人类的逐步 向 ICC 进展，伴随着表现出 CD8+ T 的免疫抑制性 TME 的积累 细胞排除。我们计划建立 MUP-uPA/NRF2Act 小鼠作为研究进化的主要模型 ICC 及其免疫抑制 TME 的研究，并寻找能够消除免疫抑制和免疫抑制的治疗方法 增强对现有 PD-(L)1 抑制剂的反应。为了实现这一目标，我们将充分描述 MUP- uPA/NRF2Act 小鼠，定义 ICC 每个阶段的转录组、表观遗传和遗传改变 进展，包括胆管反应、胆道上皮内瘤变和已形成的促纤维增生性癌。 我们还将定义肿瘤浸润淋巴细胞在各个阶段的 T 细胞受体 (TCR) 库。 恶性进展并挖掘癌细胞产生介导因子的转录组数据 结缔组织增生、免疫抑制和 CD8+ T 细胞排除。这些研究将得到补充 免疫表型分析和免疫耗竭实验，其目标是鉴定可靶向分子 负责建立 ICC 特异性免疫抑制 TME 的开关。我们将使用这些信息 确定临床批准的治疗方法，可以克服免疫抑制并大大提高疗效 基于 PD-(L)1 抑制剂的 ICI 治疗。