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: 10196181
• 负责人: Richard A. Flavell
• 金额: $ 24.08万
• 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/10196181
• 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 项目总结/摘要 酒精性肝病（ALD）是美国肝脏相关死亡的第一原因。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ALD的全部谱。 ALD。在人类ALD中，存在显著的炎症、肝损伤和脂肪变性。另一方面，老鼠 ALD模型显示非常温和的表型。因此，迫切需要开发一种临床治疗方法，但仍未得到满足 相关的ALD模型，可以捕捉人类疾病的关键特征。我们已经开发了一个 人源化鼠系统，其中小鼠通过敲入人基因在关键基因座处被人源化 （MISTRG-6-Fah-KO小鼠），并且已经通过植入人Fah-KO小鼠在细胞水平上进一步人源化。 肝细胞和CD 34+干细胞。这些小鼠支持人肝细胞、免疫、内皮和 星状细胞群。重要的是，我们已经表明，人源化MISTRG-6-Fah-KO小鼠的治疗 Lieber-DeCarli乙醇液加单次饮酒引起的肝脏炎症和肝损伤更高 比非人源化的要多。因此，我们假设人体细胞在体内暴露于酒精可以更好地 模仿人类的ALD病理学。为了验证这一假设，我们建议治疗人源化MISTRG- 6-Fah-KO小鼠进行酒精饮食，并与人类相比检查疾病病理学。接下来我们就 检查MISTRG-6-Fah-KO小鼠的人肝细胞的转录状态和组成， 酒精治疗以及它们与人类ALD的重叠程度。与拟议 我们的目标是开发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 将驱动炎症和肝损伤的发展，而人类肝细胞将驱动炎症和肝损伤的发展。 脂肪变性