CFTR modulates innate immune response in biliary epithelium: Role in the pathogenesis and treatment of Cystic-Fibrosis-related liver disease.

CFTR 调节胆管上皮的先天免疫反应：在囊性纤维化相关肝病的发病机制和治疗中的作用。

• 批准号: 9982301
• 负责人: Mario Strazzabosco
• 金额: $ 56.09万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9982301
• 关键词: Affect; Bicarbonates; Bile Acids; Bile fluid; Biliary; Biliary cirrhosis; Biochemical; Birth; Cells; Chlorides; Chronic; Clinical; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Sequence Alteration; Data; Delta F508 mutation; Development; Duct (organ) structure; Endotoxemia; Endotoxins; Epithelial; Epithelial Physiology; Epithelium; Event; Exposure to; Family; Fluids and Secretions; Foundations; Funding; Genetic; Genetic Diseases; Hepatobiliary; Human; Impairment; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Innate Immune Response; Innate Immune System; Intervention; Intestinal permeability; Intestines; Knockout Mice; Knowledge; Ligands; Liver; Liver diseases; Mediating; Membrane Proteins; Modeling; Mus; Mutation; Natural Immunity; Noxae; Outcome; PTPRC gene; Pathogenesis; Patients; Phenotype; Phosphotransferases; Play; Prevalence; Production; Quality of life; Regulation; Role; Sclerosing Cholangitis; Signal Transduction; TLR4 gene; Testing; acquired factor; apical membrane; base; bile duct; biliary tract; cholangiocyte; clinically significant; cystic fibrosis mouse; cystic fibrosis patients; cytokine; dextran sulfate sodium induced colitis; disease-causing mutation; experimental study; gut microbiota; improved; induced pluripotent stem cell; innate immune mechanisms; liver cystic fibrosis; liver development; liver transplantation; macrophage; microbiota; novel; response; src-Family Kinases; stem cell technology; treatment strategy

SUMMARY Cystic Fibrosis (CF) is a common and clinically severe genetic disease, caused by mutations in CFTR, a membrane protein that mediates chloride and fluid secretion in a number of secretory epithelia, including the biliary tree. Cystic Fibrosis liver disease (CFLD) is a chronic cholangiopathy that can eventually evolve into sclerosing cholangitis and focal biliary cirrhosis. The pathogenesis of this condition is not well understood and treatment is limited to the administration of choleretic bile acids, or in selected cases, liver transplantation. CFLD has been classically considered a consequence of the impaired bile secretion caused by the defective CFTR channel function. However, while biliary secretion is universally reduced in CF, the spontaneous development of CFLD is less frequent, suggesting that genetic and/or acquired factors are at play. Our previous studies suggested that reduced tolerance of the biliary innate immune system to endotoxins plays a major pathogenetic role in CFLD. We showed that cholangiocytes isolated from Cftr-KO mice have higher NF-κB activity and secrete a larger amount of inflammatory cytokines, when exposed to the TLR4-ligand LPS. We have also demonstrated that in CF-defective cholangiocytes, TLR4 is activated by the unrestrained function of Src family kinases (SFK), a consequence of defective CFTR. This mechanism is present also in cholangiocytes derived from human iPSC homozygous for the ΔF508 mutation. In addition, novel exciting preliminary data show that the gut microbiota in CFTR-KO mice is already different from WT littermates at birth and it is skewed towards the prevalence of a more pro-inflammatory flora. In this application we will test the hypothesis that CFLD may result from the combination of a genetic mutation affecting biliary epithelial innate immunity along with changes in microbiota composition and increased intestinal permeability. In particular, (1) we will use iPSC technology to dissect the impact of functionally different CFTR mutations on the mechanisms leading to a pro-inflammatory phenotype in human cholangiocytes and (2) we will study whether changes in the gut microbiota play a causal role in the development of liver disease in mouse models of CF. Our study will discover novel aspects of secretory epithelia physiology and innate immunity and clarity if changes in the gut microbiota play a possible causal role in CFLD. These studies represent a paradigm-shift in the understanding of the pathogenesis of CFLD and imply that treatment for CFLD should also control inflammation and the impact of the intestinal microbiota. The outcome of this project will lay the foundation for novel intervention strategies that will have an impact on the management of CFLD and other cholangiopathies. !

概括 囊性纤维化（CF）是一种常见且临床上严重的遗传疾病，是由CFTR突变引起的 在许多秘书上皮中介导氯化物和液体分泌的膜蛋白，包括 胆汁树。囊性纤维化肝病（CFLD）是一种慢性胆管疾病，有时可以演变成 硬化性胆管炎和局灶性胆汁肝硬化。这种情况的发病机理尚不清楚，并且 治疗仅限于胆碱胆酸或在某些情况下肝移植的施用。 在经典上，CFLD被认为是胆汁分泌受损的结果 有缺陷的CFTR通道功能。但是，尽管CF普遍降低了胆道的分泌 CFLD的赞助开发较少，表明遗传和/或获得因素是 玩。我们先前的研究表明，胆道先天免疫系统对内毒素的耐受性降低 在CFLD中起主要的致病作用。我们表明，从CFTR-KO小鼠中分离出的胆管细胞具有 当暴露于TLR4配体时 LPS。我们还证明，在CF缺陷性胆管细胞中，TLR4被无情的 SRC家族激酶（SFK）的功能，CFTR有缺陷的结果。这种机制也存在于 源自ΔF508突变的人IPSC纯合的胆管细胞。此外，令人兴奋的新颖 初步数据表明，CFTR-KO小鼠中的肠道微生物群已经与出生时的wt同窝室不同 它倾向于促进促炎的菌群的流行。 在此应用程序中，我们将测试以下假设：CFLD可能是由通用的组合引起的 影响胆道上皮先天免疫的突变以及微生物群组成和变化 肠渗透性增加。特别是（1）我们将使用IPSC技术来剖析 功能上不同的CFTR突变在导致人类促炎表型的机制上 胆管细胞和（2）我们将研究肠道微生物群的变化在 CF小鼠模型中肝病的发展。 如果 肠道微生物群的变化在CFLD中起可能的因果作用。这些研究代表了 对CFLD发病机理的理解，并暗示CFLD的治疗也应控制 炎症和肠道菌群的影响。该项目的结果将为 新颖的干预策略将对CFLD和其他胆管病的管理产生影响。 呢