A human Liver-on-a-Chip model for studying alcohol-associated liver disease

用于研究酒精相关肝病的人类芯片肝脏模型

基本信息

批准号：
10752839
负责人：
EKIHIRO SEKI
金额：
$ 43.84万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-11 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10752839
关键词：
3-Dimensional Acetaldehyde Adherent Culture Alcohol dehydrogenase Alcoholic Hepatitis Alcoholic Liver Diseases Alcoholic liver damage Alcohols Aldehydes Animal Model Biological Models CYP2E1 gene Cancer cell line Cell Culture System Cell Culture Techniques Cell Death Cells Cessation of life Cirrhosis Data Development Devices Disease Disease model Endothelial Cells Engineering Enzymes Ethanol Fibrosis Health Heavy Drinking Hepatic Hepatic Stellate Cell Hepatocyte Human In Vitro Lipids Liver Liver diseases Malignant neoplasm of liver Mediating Metabolism Modeling Molecular Patients Phenotype Physiological Physiology Play Predisposition Production Reactive Oxygen Species Reporting Research Risk Factors Rodent Role Study models System Testing Tissues Variant alcohol measurement aldehyde dehydrogenases design disease phenotype effective therapy experimental study in vivo insight liver inflammation liver injury microphysiology system mortality novel senescence tool

项目摘要

Project Summary Alcohol-associated liver disease (ALD) is still a serious health concern as 47% of liver-related death is due to excessive alcohol consumption. Despite of extensive research efforts, no effective therapy has been developed for ALD. One of the reasons suspending the development of effective therapy for ALD could be the lack of model systems suitable to study human relevant molecular mechanism of ALD. The current available cell culture systems and rodent ALD models do not recapitulate an ALD condition relevant in humans. To investigate more physiological experimental conditions in vitro, we established a Liver-on-a-chip model, which is a 3D in vitro hepatic micro-physiological system designed to simulate the in vivo functional conditions of liver tissues by co-culturing human-originated cells. We recently engineered a human liver-on-a-chip model suitable for investigating the underlying molecular mechanism of ALD in vitro based on the fact that human primary hepatocytes on liver-on-a-chip express higher levels of alcohol-metabolizing enzymes, such as alcohol dehydrogenase 1 (ADH1), aldehyde dehydrogenase 2 (ALDH2), and CYP2E1, compared with monolayer cultured hepatocytes. In this proposal, based on our new preliminary data using a liver-on-a-chip model, we hypothesis that, in addition to hepatocytes, liver sinusoidal endothelial cells (LSECs) and aldehyde metabolism in LSECs play important roles in regulating ethanol-induced liver damage. We will investigate whether ALDH2 in LSECs contributes to the protection for the development of ALD through acetaldehyde metabolism using a Liver-on-a-chip system (Aim 1). Because fibrosis is a risk factor for high-mortality of alcoholic hepatitis (AH), we further hypothesize that fibrotic changes in hepatic stellate cells (HSCs) and senescent changes in LSECs contribute to the increased susceptibility of ethanol-induced liver damage. We will investigate whether senescent LSECs and fibrotic HSCs contribute to the development of the microenvironment of severe ALD in cirrhotic livers as the mechanism of the development of severe AH in cirrhosis using a Liver-on-a-chip system (Aim 2). The proposed study will provide new insight into the role of ALDH2 expression and acetaldehyde metabolism in various liver cells in the progression of ethanol-induced liver damage, and provide the basis for the use of a liver-on-a-chip model as a new in vitro experimental tool to study ALD with human relevant conditions.

项目概要酒精相关性肝病 (ALD) 仍然是一个严重的健康问题，因为 47% 的肝脏相关死亡是由于过量饮酒。尽管进行了大量的研究工作，但尚未找到有效的治疗方法为 ALD 开发。暂停开发 ALD 有效疗法的原因之一可能是缺乏适合研究人类ALD相关分子机制的模型系统。目前可用的细胞培养系统和啮齿动物 ALD 模型不能概括与人类相关的 ALD 状况。到为了研究更多的体外生理实验条件，我们建立了肝脏芯片模型，是一个3D体外肝脏微生理系统，旨在模拟肝脏的体内功能状况通过共培养人类来源的细胞来形成组织。我们最近设计了一个适合的人类肝脏芯片模型基于人类原发性 ALD 的事实，在体外研究 ALD 的潜在分子机制肝脏芯片上的肝细胞表达更高水平的酒精代谢酶，例如酒精脱氢酶 1 (ADH1)、醛脱氢酶 2 (ALDH2) 和 CYP2E1，与单层细胞相比培养的肝细胞。在这项提案中，基于我们使用肝脏芯片模型的新初步数据，我们假设除了肝细胞外，肝窦内皮细胞（LSEC）也与醛代谢有关 LSEC 在调节乙醇引起的肝损伤中发挥重要作用。我们将调查是否 LSEC 中的 ALDH2 通过乙醛有助于保护 ALD 的发展使用肝脏芯片系统进行新陈代谢（目标 1）。因为纤维化是高死亡率的危险因素酒精性肝炎（AH），我们进一步假设肝星状细胞（HSC）和 LSEC 的衰老变化导致乙醇引起的肝损伤的易感性增加。我们将研究衰老的 LSEC 和纤维化的 HSC 是否有助于肝硬化严重ALD的微环境是严重AH发生的机制使用肝脏芯片系统治疗肝硬化（目标 2）。拟议的研究将为我们的作用提供新的见解乙醇诱导的肝损伤进展中各种肝细胞中 ALDH2 的表达和乙醛代谢肝损伤，并为使用肝脏芯片模型作为新的体外实验工具提供基础研究人类相关条件下的 ALD。