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）是一种重要且令人困惑的新生儿疾病数十年来，病因和病理生理学的发现。最近，NIDDK支持儿童网络在患有心脏和心脏和腹部侧向特征 - 与BA脾脏畸形（BASM）综合征的腹部特征确定该群体中是否有多器官发育的遗传病因畸形发生。 BASM外显子组序列的分析发现了几个参与者睫状基因PKD1L1的显着突变，一种与心脏侧向缺陷相关的基因，但尚未与胆道疾病有关。为了探索机械后果人类中的PKD1L1信号受损，我们开发了ep骨内胆管细胞受限的 PKD1L1FL/FL; AFP-CRE（LKO）鼠标。初步数据表明在发育小鼠肝脏不仅导致早期胆汁畸形症，而且会导致增强的周长。成人年龄的肌膜炎症，在胆管连接后不同目标的情况下（BDL）。这些组织学特征非常模仿了人类BA生活中的那些组织学特征。 AIM 1探索 LKO和其他信息中缺乏PKD1L1信号传导的纤维炎症后果 PKD1L1FL/FL杂交线（包括一个带有人类胆汁池的pkd1fl/fl PKD1L1在整个胆道中）和对选择基于胆汁的治疗干预措施的反应。 AIM 2探讨了早期胆管畸形学在发展产前和早期的描述产后生活在谱系跟踪和多重空间RNA研究中。最后，AIM 3是胆管细胞器官，偏光式植物培养物和3D导管的体外实验集片上的研究定义了分离的pkd1l1fl/fl中的分子和信号传导后果 LKO胆管细胞。总之，我们预计这三个目标将提供有史以来第一个遗传 BA模型中毒，发现了胆道反应性的新细胞和分子机制和损坏。另外，在信息丰富的PKD1L1小鼠中测试基于胆汁酸途径的剂模型可能有助于提供支持性的临床前证据，以解决当前的缺乏有效的医疗疗法。