Modeling genetic contributions to biliary atresia

模拟遗传对胆道闭锁的影响

基本信息

批准号：
10639240
负责人：
SAUL J. KARPEN
金额：
$ 64.01万
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：
2023-06-15 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10639240
关键词：
3-Dimensional Abdomen Address Adult Age Agonist Antibodies Apical Bile Acids Bile fluid Biliary Biliary Atresia Biliary Tract Diseases Biology Cardiac Cell physiology Cells Chemistry Child Child Support Cholestasis Cilia Clinical Comparative Study Crossbreeding Data Defect Development Disease Distal Duct (organ) structure Dysmorphology Etiology Evaluation Exposure to Extrahepatic Bile Ducts Functional disorder Generations Genes Genetic Models Genetic Predisposition to Disease Genetic study Handedness Histologic Histology Human Human Genetics Impairment In Vitro Individual Knowledge Ligation Link Liver Medical Membrane Membrane Proteins Modeling Molecular Mus Mutation N-terminal National Institute of Diabetes and Digestive and Kidney Diseases Obstruction Organ Organoids Oryctolagus cuniculus Participant Pathogenesis Pathway interactions Polycystic Kidney Diseases Proteomics RNA Research Role Serum Signal Transduction Subcellular structure Surface Syndrome Testing Therapeutic Therapeutic Intervention Time Transgenic Organisms Treatment Efficacy Variant bile duct biliary tract cholangiocyte efficacy evaluation exome exome sequencing experimental study hydrophilicity inhibitor insight intrahepatic liver development liver transplantation malformation mouse model neonate novel novel strategies postnatal pre-clinical prenatal pressure response three-dimensional modeling transcriptomics

项目摘要

Project Summary/Abstract Biliary atresia (BA) is an important and perplexing disease of neonates that has eluded major discoveries of etiology and pathophysiology for decades. Recently, the NIDDK-supported ChiLDReN network performed exome sequencing on a subset of BA individuals with cardiac and abdominal laterality features–those with the BA Splenic Malformation (BASM) syndrome in order to determine if there is a genetic etiology in this group with multi-organ developmental dysmorphogenesis. Analysis of BASM exome sequences found several participants with significant mutations in the ciliary gene PKD1L1, a gene associated with cardiac laterality defects, but not yet linked to biliary tract disease. In order to explore mechanistic consequences to impaired PKD1L1 signaling in humans, we developed an intrahepatic cholangiocyte-restricted Pkd1l1Fl/Fl;Afp-Cre (LKO) mouse. Preliminary data indicates that absence of Pkd1l1 in the developing mouse liver leads not only to early biliary dysmorphology, but an 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. Aim 1 explores the fibroinflammatory consequences of absent Pkd1l1 signaling in the LKO and other informative Pkd1l1Fl/Fl cross-bred lines (including one with a human bile acid pool and another that will delete Pkd1l1 in the entire biliary tree) and response to select bile acid based therapeutic interventions. Aim 2 explores the delineation of early bile duct dysmorphology in developing prenatal and early postnatal livers with lineage tracing and multiplexed spatial RNA studies. Finally, Aim 3 is an in vitro set of experiments with cholangiocyte organoids, polarized Transwell cultures and 3d duct- on-a-chip studies to define the molecular and signaling consequences in isolated Pkd1l1Fl/Fl and LKO cholangiocytes. Taken together we anticipate that these 3 Aims will provide first-ever genetic models of BA poised to discover new cellular and molecular mechanisms of biliary tract reactivity and damage. In addition, testing of bile acid pathway-based agents in informative Pkd1l1 mouse models may help provide supportive pre-clinical evidence to address the current paucity of effective medical therapeutics in BA.

项目概要/摘要胆道闭锁（BA）是一种重要且令人困惑的新生儿疾病，目前尚未引起人们的重视。最近，NIDDK 支持了数十年的病因学和病理生理学发现。 ChiLDReN 网络对患有心脏病和心脏病的 BA 个体子集进行了外显子组测序后来的腹部特征——按顺序患有 BA 脾畸形 (BASM) 综合征的人以确定该多器官发育群体是否存在遗传病因 BASM 外显子组序列的分析发现了一些异常发生。睫状基因 PKD1L1 发生显着突变，该基因与心脏偏侧性缺陷相关，但尚未与胆道疾病联系起来，以探索其机制后果。针对人类 PKD1L1 信号传导受损，我们开发了一种肝内胆管细胞限制性 Pkd1l1Fl/Fl;Afp-Cre (LKO) 小鼠的初步数据表明 Pkd1l1 缺失。发育中的小鼠肝脏不仅导致早期胆道畸形，而且导致胆道周围功能增强成人纤维炎症，尤其是胆管结扎后远端梗阻的情况 (BDL)。这些组织学特征与人类 BA 肝脏中观察到的特征非常相似。 LKO 中缺乏 Pkd1l1 信号传导的纤维炎症后果和其他信息 Pkd1l1Fl/Fl 杂交品系（包括一个带有人类胆汁酸库的品系和另一个会删除整个胆管树中的 Pkd1l1）以及对选择基于胆汁酸的治疗干预措施的反应。目标 2 探讨产前和早期发育过程中早期胆管畸形的描述最后，Aim 3 是一项具有谱系追踪和多重空间 RNA 研究的产后肝脏研究。使用胆管细胞类器官、极化 Transwell 培养物和 3d 导管进行体外实验片上研究，以确定分离的 Pkd1l1Fl/Fl 和综上所述，我们预计这 3 个目标将提供有史以来第一个遗传基因。 BA 模型有望发现胆道反应性的新细胞和分子机制此外，在信息丰富的 Pkd1l1 小鼠中测试基于胆汁酸途径的药物。模型可能有助于提供支持性临床前证据，以解决目前缺乏的问题 BA 中有效的药物治疗。