Generation and characterization of a humanized mouse model of alcoholic liver disease

酒精性肝病人源化小鼠模型的生成和表征

• 批准号: 10403562
• 负责人: Richard A. Flavell
• 金额: $ 19.89万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-10 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10403562
• 关键词: Acute; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcoholic liver damage; Alcohols; Animal Disease Models; Animal Model; CD34 gene; Cells; Cessation of life; Cholestasis; Clinical; Data; Development; Diet; Disease; Disease model; Endothelial Cells; Endothelium; Engraftment; Environment; Ethanol; Etiology; Fatty Liver; Fumarylacetoacetase; Future; Generations; Genes; Genetic Transcription; Granulocyte-Macrophage Colony-Stimulating Factor; Hepatocyte; Human; Human Pathology; IL3 Gene; Immune; Individual; Inflammation; Inflammatory; Injury; Interleukin-6; Knock-in; Knockout Mice; Left; Liquid substance; Liver; Macrophage Colony-Stimulating Factor; Modeling; Mus; Names; National Institute on Alcohol Abuse and Alcoholism; Nuclear Pore Complex; Pathology; Pathway interactions; Phenotype; Population; Proteins; Syndrome; System; Testing; Transcriptional Activation; Umbilical Cord Blood; alcohol abuse therapy; alcohol response; cell type; clinically relevant; cytokine; effective therapy; experimental study; exposed human population; human data; human disease; humanized mouse; in vivo; in vivo Model; liver inflammation; liver injury; mortality; mouse model; neutrophil; problem drinker; response; response to injury; single-cell RNA sequencing; stellate cell; stem cells; transcriptome sequencing

Generation and characterization of a humanized mouse model of Alcoholic liver disease Project Summary/Abstract Alcoholic Liver Disease (ALD) is the first cause of liver related deaths in USA. However, effective treatment options for ALD are very limited due to the lack of suitable in vivo models that recapitulate the full spectrum of ALD. In human ALD, there is marked inflammation, liver damage and steatosis. On the other hand, mouse models of ALD display very mild phenotype. Thus, there is an urgent but still unmet need to develop a clinical relevant ALD model that can capture the key features of human disease. We have already developed a humanized murine system in which mice have been humanized at key loci by knock-in of human genes (MISTRG-6-Fah-KO mouse) and have been further humanized at a cellular level by engraftment of human hepatocytes and CD34+ stem cells. These mice support human liver hepatocytes, immune, endothelial and stellate cell populations. Crucially, we have shown that the treatment of humanized MISTRG-6-Fah-KO mice with Lieber-DeCarli ethanol liquid plus a single binge ethanol induced higher liver inflammation and liver damage than in non-humanized. Therefore, we hypothesize that the exposure of human cells to alcohol in vivo can better mimic the ALD pathology of humans. To test this hypothesis, we are proposing to treat the humanized MISTRG- 6-Fah-KO mice with alcohol diet and to examine disease pathology in comparison to humans. Next, we will examine the transcriptional state and the composition of human liver cells of MISTRG-6-Fah-KO mice upon alcohol treatment and how much is the extent of their overlap with the human ALD. With the proposed experiments, we aim to develop and characterize the first human-clinical relevant alcoholic liver damage model in the humanized MISTRG-6-Fah-KO mice. In that model, human immune cells and human effector cytokines will drive the development of inflammation and liver damage and human hepatocytes will drive the development of steatosis.

人源化酒精性肝病小鼠模型的建立及鉴定 项目摘要/摘要 酒精性肝病(ALD)是美国肝脏相关死亡的第一位原因。然而，有效的治疗 ALD的选择非常有限，因为缺乏合适的体内模型来概括全谱的ALD ALD.在人类ALD中，存在明显的炎症、肝脏损伤和脂肪变性。另一方面，鼠标 ALD模型表现出非常轻微的表型。因此，有一种迫切但仍未得到满足的需求，即开发一种临床 相关的ALD模型，可以捕捉人类疾病的关键特征。我们已经开发了一种 通过敲入人类基因使小鼠在关键位置人源化的人源化小鼠系统 (MISTRG-6-Fah-KO小鼠)，并通过植入人类在细胞水平上进一步人性化 肝细胞和CD34+干细胞。这些小鼠支持人肝细胞、免疫、内皮和 星状细胞群。至关重要的是，我们已经证明了人源化MISTRG-6-Fah-KO小鼠的治疗 Lieber-DeCarli酒精液加单次狂饮酒精引起更高的肝脏炎症和肝损伤 而不是非人性化的。因此，我们假设人体细胞在体内暴露在酒精中可以更好地 模仿人类的阿尔茨海默病的病理。为了验证这一假设，我们建议将人性化的MISTRG- 6-Fah-KO小鼠给予酒精饮食，并与人类比较检查疾病病理。接下来，我们将 检测MISTRG-6-Fah-KO小鼠的人肝细胞转录状态和组成 酒精治疗以及它们与人类ALD重叠的程度是多少。与建议的 通过实验，我们的目标是建立和表征第一个人类临床相关的酒精性肝损伤模型。 在人源化的MISTRG-6-Fah-KO小鼠中。在该模型中，人类免疫细胞和人类效应细胞因子 将推动炎症和肝脏损伤的发展，而人类肝细胞将推动发展 脂肪变性。