Roles for Pkd1l1 in bile duct development

Pkd1l1 在胆管发育中的作用

基本信息

批准号：
10751883
负责人：
dominick Hellen
金额：
$ 4.77万
依托单位：
EMORY UNIVERSITY
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：
2023-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10751883
关键词：
1 year old Abnormal Cell Address Adult Age Agonist Alleles Antibodies Basic Science Bile Acids Biliary Biliary Atresia Biliary cirrhosis Biology Cell Communication Cell Separation Cell physiology Cells Cellular Structures Child Cholestasis Cilia Confocal Microscopy Data Defect Detection Development Disease Distal Duct (organ) structure Dysmorphology Embryonic Development Epithelium Etiology Evaluation Extrahepatic Bile Ducts Fibrosis Functional disorder Genes Genetic Genetic Transcription Genetic study Goals Handedness Hepatology Histologic Human Immunoprecipitation Impairment In Situ In Vitro Individual Infant Intrahepatic bile duct Knock-out Knowledge Ligation Liquid substance Liver Liver diseases Medical Membrane Membrane Proteins Modeling Molecular Molecular Analysis Mus Mutation Neonatal Obstruction Organ PKD2 protein Participant Pathogenesis Pathology Phenotype Polycystic Kidney Diseases Proliferating Proteomics Publishing RNA Regulation Reporter Reporting Research Research Personnel Role Sensory Signal Transduction Solid Structural Protein Syndrome Techniques Testing Therapeutic Training Translational Research Transmission Electron Microscopy Transplantation Work bile duct biliary tract cholangiocyte exome sequencing experimental study in vivo insight intrahepatic liver transplantation malformation mouse model neonate novel postnatal pre-clinical prenatal response skills stem therapeutic target three-dimensional modeling transcriptome sequencing

1 year old Abnormal Cell Address Adult Age Agonist Alleles Antibodies Basic Science Bile Acids Biliary Biliary Atresia Biliary cirrhosis Biology Cell Communication Cell Separation Cell physiology Cells Cellular Structures Child Cholestasis Cilia Confocal Microscopy Data Defect Detection Development Disease Distal Duct (organ) structure Dysmorphology Embryonic Development Epithelium Etiology Evaluation Extrahepatic Bile Ducts Fibrosis Functional disorder Genes Genetic Genetic Transcription Genetic study Goals Handedness Hepatology Histologic Human Immunoprecipitation Impairment In Situ In Vitro Individual Infant Intrahepatic bile duct Knock-out Knowledge Ligation Liquid substance Liver Liver diseases Medical Membrane Membrane Proteins Modeling Molecular Molecular Analysis Mus Mutation Neonatal Obstruction Organ PKD2 protein Participant Pathogenesis Pathology Phenotype Polycystic Kidney Diseases Proliferating Proteomics Publishing RNA Regulation Reporter Reporting Research Research Personnel Role Sensory Signal Transduction Solid Structural Protein Syndrome Techniques Testing Therapeutic Training Translational Research Transmission Electron Microscopy Transplantation Work bile duct biliary tract cholangiocyte exome sequencing experimental study in vivo insight intrahepatic liver transplantation malformation mouse model neonate novel postnatal pre-clinical prenatal response skills stem therapeutic target three-dimensional modeling transcriptome sequencing

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项目摘要

PROJECT SUMMARY Biliary atresia (BA) is the main indication to transplant any solid organ in infants less than 1 year of age and has eluded major discoveries of etiology and pathophysiology for decades. BA is a neonatal liver disease that is best characterized by fibroinflammatory obstruction of both intra- and extrahepatic bile ducts. Through exome sequencing of a subset of BA individuals with heterotaxic (laterality) features (those with the BA Splenic Malformation syndrome), several participants were identified with biallelic damaging mutations in the ciliary gene PKD1L1 (Polycystic kidney disease 1 like 1). To explore mechanistic consequences of impaired PKD1L1 signaling in humans, we developed an intrahepatic cholangiocyte-restricted Pkd1l1Fl/Fl;Afp-Cre (LKO) mouse. Recently published data indicates that the absence of Pkd1l1 in the developing mouse liver leads to early biliary dysmorphology and enhanced peribiliary fibroinflammation at adult ages, moreso in the setting of distal obstruction after bile duct ligation (BDL). These histologic features strongly mimic those seen in human BA livers, offering a novel opportunity to discover specific cellular and molecular insights into BA’s rapid and profound pathogenesis. There are two paradigms that must be uncovered to provide a more thorough understanding of Pkd1l1, and thus the molecular pathogenesis of BA. First, the consequences of absent Pkd1l1 in the developing biliary tree that leads to adulthood biliary fibroinflammation are unknown. Second, the mechanism of Pkd1l1 signaling within cholangiocytes, which when absent contributes to biliary pathology, remains to be elucidated. The overarching hypothesis in this proposal is that Pkd1l1 is required for proper biliary development and signaling. This hypothesis will be tested through the following two aims. Aim 1 explores the delineation and characterization of early bile duct dysmorphology in developing prenatal and early postnatal livers. This aim will utilize the recently developed Pkd1l1Null/Fl (NullFl) and Pkd1l1Null/Fl;Afp-cre (NullLKO) mouse lines. Aim 2 is an in vitro set of experiments with isolated cholangiocyte studies to define Pkd1l1-interactors and signaling consequences, specifically in response to various bile acids, in isolated Pkd1l1Fl/Fl and LKO cholangiocytes. Taken together, we anticipate that these two aims will discover new cellular and molecular mechanisms of biliary tract development and signaling. In addition, information stemming from these Pkd1l1-based mouse models will help provide supportive pre-clinical evidence to address the current paucity of effective medical therapeutics in BA.

项目摘要胆道闭锁（BA）是在不到1岁的婴儿中移植任何固体器官的主要指标，并且具有数十年来，躲避病因和病理生理学的主要发现。 BA是一种最佳的新生儿肝病其特征在于脑内和外膜外胆管的纤维炎症反对。通过外显子具有异质性（侧向）特征的BA个体的测序（具有BA脾畸形综合征），在睫状基因中发现了几名参与者的生物损害突变 PKD1L1（多囊肾脏病1类似1）。探索受损PKD1L1的机械后果我们在人类中的信号传导，开发了一种e脚内胆管细胞限制的PKD1L1FL/FL; AFP-CRE（LKO）小鼠。最近发表的数据表明，发育中的小鼠肝脏中缺乏PKD1L1导致早期胆道成年时代的畸形和增强的周围肌膜炎症，在不同的环境中胆管连接后的阻塞（BDL）。这些组织学特征非常模仿人类肝中看到的那些组织学特征，提供了一个新的机会，可以发现特定的细胞和分子见解，以了解BA的快速而深刻发病。必须发现两个范式，以提供对 PKD1L1，因此Ba的分子发病机理。首先，发育中缺乏PKD1L1的后果导致成年胆道肌炎症的胆道未知。第二，PKD1L1的机制胆管细胞内的信号传导，如果没有造成胆道病理的影响，仍有待阐明。该提议中的总体假设是PKD1L1是适当的胆道开发所必需的和信号。该假设将通过以下两个目标进行检验。 AIM 1探索描述以及早期胆管畸形学在发展产后和早期生命中的表征。这个目标将利用最近开发的PKD1L1NULL/FL（NULLFL）和PKD1L1NULL/FL; AFP-CRE（NULLLKO）小鼠线。 AIM 2是一组孤立的胆管细胞研究的体外实验，以定义PKD1L1相互作用和信号传导后果，特别是针对各种胆汁酸的响应，在分离的PKD1L1FL/FL和LKO胆管细胞中的后果。综上所述，我们预计这两个目标将发现胆道的新细胞和分子机制道的发展和信号传导。此外，这些基于PKD1L1的鼠标模型的信息将帮助提供支持性的临床前证据，以解决当前有效医疗疗法的缺乏 BA。