Improving hepatocellular carcinoma mouse modeling by understanding the malignant potential and biology of liver cell subpopulations

通过了解肝细胞亚群的恶性潜能和生物学来改善肝细胞癌小鼠模型

• 批准号: 10610474
• 负责人: Hao Zhu
• 金额: $ 53.22万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10610474
• 关键词: Address; Architecture; Biological; Biological Assay; Biology; Cancer Etiology; Cause of Death; Cells; Cessation of life; Chemicals; Chronic; Cirrhosis; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Competence; Detection; Development; Etiology; Event; Frequencies; Gene Expression; Genes; Hepatic; Hepatic lobule; Hepatocyte; Heterogeneity; Homeostasis; Hypoxia; Incidence; Injury; Knock-in; Label; Liver; Liver Stem Cell; Maintenance; Malignant - descriptor; Malignant Epithelial Cell; Malignant Neoplasms; Metabolic; Metabolic Diseases; Metabolism; Methods; Modeling; Molecular; Mouse Strains; Mus; Mutate; Nutritional; Outcome; Overnutrition; Pathway interactions; Patients; Ploidies; Population; Predisposition; Prevention; Primary carcinoma of the liver cells; Reagent; Regenerative capacity; Source; Therapeutic; Tumor Biology; United States; carcinogenesis; cell type; chronic liver injury; clinically relevant; experimental study; genetic manipulation; genome editing; improved; innovation; insight; liver cancer model; liver injury; mortality; mouse model; neoplastic; nonalcoholic steatohepatitis; novel; progenitor; regenerative; regenerative cell; success; tool; tumor; tumor initiation

PROJECT SUMMARY Hepatocellular carcinoma (HCC) is the fastest growing cause of cancer-related death in the United States and one of the leading causes of death in patients with cirrhosis – the end result of any chronic liver injury. HCC is difficult to treat and outcomes have barely improved over the last 30 years, with 5-year survival under 20% and an incidence-to-mortality ratio near 1. Innovative strategies for detection, prevention, and treatment for HCC are desperately needed, the development of which will depend heavily on mouse models of liver cancer. The current models are inadequate because they do not take into account the cells of origin, which will likely have a significant impact on tumor initiation, maintenance, and therapeutic vulnerabilities. While the liver is appreciated for being a central metabolic hub and for its astounding regenerative capacity, cellular heterogeneity in the liver is mostly unexplored. Along the portal to central axis within the hepatic lobule, profound differences in gene expression, metabolism, hypoxia, and ploidy are observed. Whether or not these differences reflect differences in neoplastic potential, and whether or not they influence metabolic disease or carcinogenesis, is unclear. In recent years, there has been intense controversy about whether or not there is a liver stem/progenitor cell. To compound these debates, the injury assays used to drive HCC development often do not model common etiologies such as non- alcoholic steatohepatitis (NASH), which is emerging as the most common cause of cirrhosis in the U.S. We posit that a critical problem for HCC modeling is a lack of understanding of how different cell types contribute to cancer in the context of clinically relevant injuries. We believe that identifying the specific cellular subtypes that give rise to HCC and then being able to genetically perturb these cells is critical for us to better model HCC. Unfortunately, the field does not have mouse reagents to manipulate much of the heterogeneity in the liver. To address this, my lab has optimized CRISPR genome-editing methods to rapidly generate lineage tracing mice. Nine new CreER knock-in models that label different zone-specific and progenitor populations have been produced, effectively quadrupling the number of CreER lines available to our field. We will use these tools to trace and genetically manipulate cell types in a systematic fashion in order to identify the most important regenerative cell populations (Aim 1) and the HCC cell(s) of origin (Aim 2) in the context of clinically-relevant chemical and nutritional injuries. We will then ask if cell type specific gene manipulation of common HCC driver genes will help to uncover different transformation competencies between hepatocyte subpopulations (Aim 3). Success in this project will provide the community with a large panel of important CreER tools, allow the field to focus on important subpopulations that are more likely to transform, and reveal pathways that control tumor development in these cells.

项目概要 肝细胞癌 (HCC) 是美国癌症相关死亡增长最快的原因 肝硬化患者死亡的主要原因之一——任何慢性肝损伤的最终结果都是。 过去 30 年来，该病难以治疗，结果几乎没有改善，5 年生存率低于 20%， 发病率与死亡率之比接近 1。HCC 检测、预防和治疗的创新策略是 迫切需要，其发展将在很大程度上取决于目前的肝癌小鼠模型。 模型是不充分的，因为它们没有考虑起源细胞，而起源细胞可能具有显着的影响 肝脏因其对肿瘤发生、维持和治疗脆弱性的影响而受到赞赏。 作为一个中央代谢中心，由于其惊人的再生能力，肝脏中的细胞异质性主要是 沿着肝小叶内的门户到中轴，基因表达的深刻差异， 观察代谢、缺氧和倍性是否这些差异反映了肿瘤的差异。 近年来，尚不清楚它们是否会影响代谢疾病或致癌作用。 关于是否存在肝干/祖细胞来复合这些一直存在激烈争议。 争论中，用于驱动 HCC 发展的损伤测定通常不能模拟常见病因，例如非 酒精性脂肪性肝炎 (NASH)，它正在成为美国肝硬化的最常见原因。我们认为 HCC 建模的一个关键问题是缺乏对不同细胞类型如何导致癌症的了解 我们认为，在临床相关损伤的背景下，可以识别引起的特定细胞亚型。 不幸的是，能够从基因上扰乱这些细胞对于我们更好地模拟 HCC 至关重要。 该领域没有小鼠试剂来操纵肝脏中的大部分异质性。 我的实验室优化了 CRISPR 基因组编辑方法，可以快速生成 9 只新的谱系追踪小鼠。 已经产生了标记不同区域特异性和祖细胞群的 CreER 敲入模型， 我们将使用这些工具来跟踪和有效地将我们领域可用的 CreER 线路数量增加四倍。 以系统的方式对细胞类型进行基因操作，以识别最重要的再生细胞 人群（目标 1）和 HCC 细胞来源（目标 2）在临床相关的化学和 然后我们将询问常见 HCC 驱动基因的细胞类型特异性基因操作是否会有帮助。 揭示肝细胞亚群之间不同的转化能力（目标 3）。 项目将为社区提供大量重要的 CreER 工具，让该领域能够专注于 更有可能转化的重要亚群，并揭示控制肿瘤发展的途径 在这些细胞中。

项目成果

Liver homeostasis is maintained by midlobular zone 2 hepatocytes.

• DOI: 10.1126/science.abb1625
• 发表时间: 2021-02-26
• 期刊: Science (New York, N.Y.)
• 作者: Wei Y;Wang YG;Jia Y;Li L;Yoon J;Zhang S;Wang Z;Zhang Y;Zhu M;Sharma T;Lin YH;Hsieh MH;Albrecht JH;Le PT;Rosen CJ;Wang T;Zhu H
• 通讯作者: Zhu H