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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.

项目摘要/摘要胆道闭锁(BA)是新生儿的一种重要且令人困惑的疾病，一直没有得到很大的重视。几十年来病因学和病理生理学的发现。最近，NIDDK支持儿童网络对患有心脏和心脏疾病的BA患者的子集进行了外显子组测序腹部偏侧特征--依次为BA脾畸形(BASM)综合征以确定多器官发育组中是否存在遗传病因畸形发育。对BASM外显子序列的分析发现，几个参与者具有纤毛基因PKD1L1的显著突变，这是一种与心脏偏侧缺陷有关的基因，但尚未与胆道疾病有关。为了探索机械后果，在人类PKD1L1信号受损的情况下，我们开发了一种肝内胆管细胞受限的 Pkd1l1F1/Fl；AFP-Cre(LKO)小鼠。初步数据表明，Pkd1l1在发育中的小鼠肝脏不仅导致早期胆管畸形，而且胆管周围增强。成人纤维炎症，多见于胆管结扎后远端梗阻 (BDL)。这些组织学特征强烈地模仿了人类BA肝脏中的特征。目标1探索 LKO中Pkd1l1信号缺失的纤维炎症后果及其他信息 Pkd1l1F1/F1杂交品系(包括一个带有人胆汁酸池的品系和另一个将删除 Pkd1l1在整个胆管树中)以及对选择的基于胆汁酸的治疗干预的反应。目的2探讨早期胆管畸形在产前和早期发育中的作用。出生后肝脏的谱系追踪和多重空间RNA研究。最后，《目标3》是一个进军用胆管细胞器官、极化Transwell培养和3D导管进行的体外实验- 芯片上研究以确定分离的Pkd1L1F1和Fl中的分子和信号后果 LKO胆管细胞。综上所述，我们预计这3个目标将提供有史以来第一个基因 BA模型有望发现胆道反应性的新细胞和分子机制和破坏。此外，在提供信息的Pkd1l1小鼠身上测试基于胆汁酸途径的药物模型可能有助于提供支持性的临床前证据，以解决目前缺乏英国航空公司的有效医疗疗法。