Pathobiology of Hepatic Epithelia

肝上皮的病理生物学

• 批准号: 7223475
• 负责人: Nicholas F. LaRusso
• 金额: $ 50.22万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-12-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7223475
• 关键词: Address; Affect; Animal Model; Apical; Area; Autosomal Recessive Polycystic Kidney; Award; Bile Acids; Bile fluid; Biliary; Cell Line; Cell physiology; Chemicals; Cilia; Clinical; Condition; Cyst; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Disease; Ductal; Epithelial Cells; Epithelium; Experimental Animal Model; Family; Functional disorder; Gene Silencing; Genetic; Glucose; Hepatic; Hepatobiliary; Hepatocyte; Hereditary Disease; Heterogeneity; Intracellular Second Messenger; Intrahepatic bile duct; Ion Transport; Ions; Knockout Mice; Liver; Mechanics; Membrane; Methodology; Methods; Modeling; Molecular; Molecular and Cellular Biology; Mutate; Nucleotides; Organelles; PKHD1 gene; Pathologic Processes; Pathway interactions; Physiology; Production; Proteins; Publishing; Rate; Rattus; Recycling; Regulation; Research; Role; Second Messenger Systems; Sensory; Signal Transduction; Small Interfering RNA; Structure; Surface; System; Techniques; Technology; Testing; Transgenic Organisms; United States National Institutes of Health; Water; Water Movements; apical membrane; bile duct; bile formation; biliary tract; cholangiocyte; concept; innovation; insight; novel; programs; receptor; solute; water channel

DESCRIPTION (provided by applicant): Our long-term objectives remain to apply the fundamental concepts and broad technologies of cell and molecular biology to understand hepatic epithelial cell function and dysfunction. We continue to focus on cholangiocytes, the epithelial cells lining intrahepatic bile ducts, because of their biologic and clinical importance, and because of the new concepts, hypotheses, and techniques we have developed to study, cholangiocyte pathobiology, an underserved area of liver research. Recent evidence from our lab indicates that: (i) aquaporins (AQPs), a family of water channels, are important in ductal bile formation; and (ii) cholangiocytes contain primary cilia that act as sensory organelles and participate in normal bile formation and in biliary cystogenesis. Thus, we will test the central hypothesis that ductal bile formation: (i) is the net result of solute-driven, passive movement of water molecules through AQPs constitutively expressed in or recycled among distinct cellular compartments; (ii) is influenced by luminal mechanical, chemical, and osmotic signals sensed via primary cilia on the apical cholangiocyte membrane; and (iii) is abnormal in genetic spontaneous or experimental animal models of autosomal recessive polycystic kidney disease (ARPKD) when cholangiocyte ciliary structure and/or function is disturbed. Our three distinct but integrated specific aims test the hypotheses that: (i) ductal bile formation involves the normal function of primary cilia expressed on the apical membrane of each cholangiocyte to detect mechanical (e.g., bile flow rate), chemical (e.g., nucleotides, bile acids, glucose), and/or osmotic (e.g., bile hypo/hyperosmolarity) signals from bile; (ii) cellular expression, compartmentalization, and recycling of key 'flux' proteins (e.g., AQPs, AE2, CFTR) regulating ductal bile formation are influenced by ciliary stimulation; and (iii) abnormalities in structure, expression, and/or cellular localization of cilia-associated proteins (e.g., fibrocystin, the protein product of PKHD1, the gene mutated in ARPKD) contribute to disturbances in cholangiocyte water, solute, and ion transport promoting biliary cystogenesis. We will employ established and new methods, models, and probes, including: perfused bile duct units, isolated biliary cysts, isolated cholangiocyte cilia, spontaneous (i.e., the PCK rat) and transgenic (i.e., fibrocystin knockout mouse) animal models of ARPKD, gene silencing using small-interfering RNAs (siRNAs), novel cholangiocyte culture systems, and innovative morphologic techniques. Our results will further clarify the role of AQPs in cholangiocyte bile formation, address directly the potential importance of cholangiocyte cilia in ductal bile production, and explore the relationship of cholangiocyte cilia to possible disturbances of water, ion, and solute transport in biliary cystogenesis. Innovative aspects of our program include novel methodologies and animal models, and new concepts regarding the importance of cholangiocyte AQPs and cilia in ductal bile formation and biliary cystogenesis. We will generate information to yield new insights into normal cholangiocyte function, explore highly promising, selected aspects of cholangiocyte dysfunction, and continue to provide a broad theoretical framework for understanding and managing the cholangiopathies, a group of genetic and acquired hepatobiliary diseases in which the cholangiocyte is the principal target of diverse pathologic processes.

描述（由申请人提供）：我们的长期目标仍然是应用细胞和分子生物学的基本概念和广泛技术来了解肝上皮细胞功能和功能障碍。我们继续关注胆管细胞，肝内胆管的上皮细胞，因为它们在生物学和临床上的重要性，也因为我们已经开发了新的概念，假设和技术来研究胆管细胞病理生物学，这是肝脏研究的一个不足领域。我们实验室的最新证据表明：（i）水通道蛋白（AQP），一个水通道家族，在胆管胆汁形成中很重要;（ii）胆管细胞含有初级纤毛，作为感觉细胞器，参与正常胆汁形成和胆汁囊肿形成。因此，我们将测试的核心假设，即胆管胆汁形成：（i）是溶质驱动的，被动运动的水分子通过水通道蛋白的净结果组成性表达或在不同的细胞室之间回收;（ii）是由管腔机械，化学和渗透信号的影响，通过初级纤毛上的顶端胆管细胞膜上的感觉;和（iii）当胆管细胞纤毛结构和/或功能受到干扰时，在常染色体隐性多囊肾病（ARPKD）的遗传自发或实验动物模型中是异常的。我们的三个不同但综合的具体目标测试假设：（i）导管胆汁形成涉及每个胆管细胞顶膜上表达的初级纤毛的正常功能，以检测机械（例如，胆汁流速），化学（例如，核苷酸，胆汁酸，葡萄糖），和/或渗透的（例如，胆汁低/高渗透压）来自胆汁的信号;（ii）细胞表达、区室化和关键“通量”蛋白的再循环（例如，调节导管胆汁形成的AQP、AE 2、CFTR）受到纤毛刺激的影响;和（iii）纤毛相关蛋白（例如，纤维囊蛋白，PKHD 1的蛋白质产物，ARPKD中突变的基因）导致胆管细胞水、溶质和离子转运的紊乱，从而促进胆管囊形成。我们将采用已建立的和新的方法、模型和探针，包括：灌注胆管单位、孤立的胆管囊肿、孤立的胆管细胞纤毛、自发的（即，PCK大鼠）和转基因（即，纤维囊蛋白敲除小鼠）ARPKD动物模型、使用小干扰RNA（siRNA）的基因沉默、新型胆管细胞培养系统和创新的形态学技术。我们的研究结果将进一步阐明水通道蛋白在胆管细胞胆汁形成中的作用，直接解决胆管细胞纤毛在导管胆汁产生中的潜在重要性，并探讨胆管细胞纤毛与胆汁囊肿形成中水、离子和溶质转运可能紊乱的关系。我们计划的创新方面包括新的方法和动物模型，以及关于胆管细胞AQP和纤毛在胆管胆汁形成和胆管囊肿形成中的重要性的新概念。我们将产生的信息，以产生新的见解正常的胆管细胞功能，探索非常有前途的，选择方面的胆管细胞功能障碍，并继续提供一个广泛的理论框架，了解和管理的胆管疾病，一组遗传性和获得性肝胆疾病，其中胆管细胞是不同的病理过程的主要目标。