In Vitro Liver Models to Investigate the Progression of Liver Fibrosis

用于研究肝纤维化进展的体外肝脏模型

• 批准号: 10016367
• 负责人: Srivatsan Kidambi
• 金额: $ 9.98万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2021-09-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10016367
• 关键词: Alcoholic Liver Diseases; Animal Model; Antineoplastic Agents; Area; Biochemical; Biology; Biomimetics; Cell Communication; Cell physiology; Cells; Characteristics; Chronic; Chronology; Cirrhosis; Clinical; Clinical Research; Coculture Techniques; Collagen; Communication; Cues; Development; Diagnosis; Disease; Disease Progression; Endothelial Cells; Engineering; Environment; Etiology; Extracellular Matrix; Extracellular Matrix Proteins; Fibrosis; Gel; Genes; Goals; Health Care Costs; Hepatic; Hepatic Stellate Cell; Hepatitis B; Hepatitis C; Hepatocyte; Human; In Vitro; Kupffer Cells; Laboratories; Laminin; Ligands; Liver; Liver Fibrosis; Liver diseases; Maintenance; Mechanics; Mediating; Modeling; Molecular; Nebraska; Participant; Peptides; Physiological; Play; Population; Role; Signal Transduction; Signaling Molecule; Staging; System; Toxicology; Variant; Virus Diseases; Work; chronic liver disease; cost; density; in vitro Model; in vivo; innovation; insight; interdisciplinary approach; liver function; liver injury; liver metabolism; nanofiber; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; response; screening; stellate cell; therapeutic biomarker; treatment response

ABSTRACT Liver fibrosis, which results from chronic liver damage in conjunction with the accumulation of extracellular matrix (ECM) proteins, is characteristic of several chronic liver diseases. Dynamic changes to the liver microenvironment (LME) are widely recognized as a critical participant in liver fibrosis progression and therapeutic responses. LME components, including interactions between parenchymal (hepatocytes) and non- parenchymal cells (liver sinusoidal endothelial cells [LSECs], hepatic stellate cells, kupffer cells), signaling molecules (ligands-collagen, laminin), and mechanical cues from ECM, have been implicated in the progression of liver fibrosis. Stellate cells activation is the hallmark of liver fibrosis; however, the effect on hepatocytes and LSECs function has not been extensively understood. Also, the mechanisms by which the LME components regulate liver function and various signaling cascades are poorly understood, thus limiting the development of optimal diagnosis and treatment regimes for liver diseases (e.g., alcoholic liver disease, nonalcoholic fatty liver disease, non-alcoholic steatohepatitis, and hepatitis B and C). Therefore, there is a critical need to develop in vitro models that simulate the dynamic LME components and effectively study their role in liver fibrosis. To study the direct effects of LME on cell signaling, it is imperative to use in vitro liver models to simulate the fundamental complexity and dynamism of liver fibrosis and to achieve greater translational validity. The goal of this application is to use a multidisciplinary approach to develop three independent in vitro liver models to study how different LME components (hepatocytes-LSEC interactions, ECM stiffness, ligand type and density) regulate hepatocyte and LSEC function and what role these LME components play in the progression of liver fibrosis. The specific aims of the proposed study are to: 1) investigate the effect of hepatocytes-LSEC interaction(s) on hepatic function, 2) investigate how variation in stiffness alters hepatic cell function, and 3) determine the role of ligand type/density in regulating liver cell function. This work will provide a significant advancement in the ability to utilize in vitro liver models to accurately describe the liver function and metabolism in normal versus diseased states, and especially how the LME regulate the development and maintenance of liver function. Importantly, this model is innovative as it will chronologically emulate the fibrosis stage, is similar to clinical conditions, and boasts an environment that is more controlled and systematic than animal models. This project is expected to have a progressive impact on the study of liver fibrosis and related fields because the availability of a liver model that retains LME will facilitate understanding of the molecular mechanisms that underlie LME activities in mechanisms critical for the maintenance of liver biology.

抽象的 肝纤维化是由慢性肝损伤和细胞外毒素积累引起的 基质（ECM）蛋白是多种慢性肝病的特征。肝脏的动态变化 微环境（LME）被广泛认为是肝纤维化进展的关键参与者 治疗反应。 LME 组成部分，包括实质（肝细胞）和非细胞之间的相互作用 实质细胞（肝窦内皮细胞 [LSEC]、肝星状细胞、库普弗细胞）、信号传导 分子（配体-胶原蛋白、层粘连蛋白）和 ECM 的机械信号，与 肝纤维化的进展。星状细胞活化是肝纤维化的标志；然而，影响 肝细胞和 LSEC 的功能尚未得到广泛了解。此外， LME 成分调节肝功能，并且对各种信号级联反应知之甚少，因此限制了 开发肝脏疾病的最佳诊断和治疗方案（例如酒精性肝病、 非酒精性脂肪肝、非酒精性脂肪性肝炎以及乙型和丙型肝炎）。因此，有一个 迫切需要开发模拟动态 LME 成分并有效研究其成分的体外模型 在肝纤维化中的作用。为了研究LME对细胞信号传导的直接影响，必须使用体外肝脏 模型来模拟肝纤维化的基本复杂性和动态性，并实现更大的目标 翻译效度。该应用程序的目标是使用多学科方法来开发三个 独立的体外肝脏模型来研究不同的 LME 成分（肝细胞 - LSEC 相互作用， ECM 硬度、配体类型和密度）调节肝细胞和 LSEC 功能以及这些 LME 的作用 成分在肝纤维化的进展中发挥作用。拟议研究的具体目标是：1) 研究肝细胞-LSEC 相互作用对肝功能的影响，2) 研究肝细胞与 LSEC 相互作用的变化如何 硬度改变肝细胞功能，3) 确定配体类型/密度在调节肝细胞中的作用 功能。这项工作将在利用体外肝脏模型的能力方面取得重大进展 准确地描述正常状态与患病状态下的肝功能和代谢，特别是如何 LME 调节肝功能的发育和维持。重要的是，这种模式具有创新性，因为它 将按时间顺序模拟纤维化阶段，与临床状况相似，并拥有一个环境 比动物模型更具控制性和系统性。该项目预计将产生积极影响 肝纤维化及相关领域的研究，因为保留 LME 的肝脏模型的可用性将 促进对 LME 活动背后的分子机制的理解，这些机制对 LME 至关重要 维持肝脏生物学。