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）是一种侵略性的原发性肝癌（PLC），特别难以进行 治疗，总5年生存率为8％。像主要PLC一样，肝细胞癌（HCC），ICC 由于胆固性肝病，非酒精性脂肪肝疾病的持续增加，发病率正在上升 2型糖尿病，酒精和烟草滥用。与HCC不同，经常在高级阶段检测到ICC 生存前景很差，这是其与早期HCC不准确分化的并发症 技术。由于其独特的管理和缺席，两个PLC的早期差异诊断至关重要 ICC和HCC共享的可靶向致癌驱动器。可以通过识别PLC-可以解决此问题 型不可知治疗方法，例如免疫检查点抑制（ICI），适用于两个ICC 和HCC。但是，尽管基于ICI的疗法已被批准用于HCC，并且其功效已得到提高 结合VEGF抑制剂，它们在ICC中的表现不佳。这种故障可以部分归因于 对ICC特异性免疫抑制肿瘤微环境（TME）和 缺乏合适的鼠标模型，可以将ICI与能够结合使用的ICI评估 拆除免疫抑制。我们通过开发一种新的ICC模型来解决此问题，该模型与 以前的小鼠模型不依赖于迅速的有效致癌驱动器的过度表达 在没有选择性压力的情况下诱导ICC来获得其他突变。我们的混乱/nrf2act 模型取决于ER应力与双重电位中转录因子NRF2激活的组合 肝胆管祖细胞和成熟的肝细胞，表现出健壮的，高度渗透，类似于人类的逐步 向ICC的进展，伴随着表现出CD8+ T的免疫抑制TME的积累 细胞排除。我们计划建立混乱/NRF2ACT小鼠作为研究进化的主要模型 ICC及其免疫抑制性TME，并寻找可以消除免疫抑制和的治疗方法 增强对现有PD-（L）1抑制剂的反应。为了实现这一目标，我们将充分表征mup- UPA/NRF2ACT小鼠，定义ICC每个阶段的转录组，表观遗传和遗传变化 进展，包括导管反应，胆道上皮内肿瘤和建立的脱肿瘤癌。 我们还将定义在每个阶段的肿瘤浸润淋巴细胞的T细胞受体（TCR）曲目 恶性进展并挖掘癌细胞产生介导的因子的转录组数据 去乳突，免疫抑制和CD8+ T细胞排除。这些研究将得到补充 免疫表型和免疫启动实验的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。我们将使用此信息 确定可以克服免疫抑制并大大提高功效的临床批准治疗方法 基于PD-（L）1抑制剂的ICI治疗。