Endothelial Cell Reprogramming in Familial Intracranial Aneurysm

家族性颅内动脉瘤的内皮细胞重编程

基本信息

批准号：
10595404
负责人：
Tanyeri Barak
金额：
$ 60.3万
依托单位：
YALE 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：
2023-02-01 至 2028-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10595404
关键词：
3-Dimensional Address Adult Affect Angiography Animal Model Animals Arteries Binding Biochemical Blood Vessels Brain Brain hemorrhage Candidate Disease Gene Cell Reprogramming Cell model Cell physiology Cells Central Nervous System Cerebral hemisphere hemorrhage Cerebrovascular system Cerebrum Code Complex Data Defect Development Diagnosis Diagnostic Disease Endothelial Cells Endothelium Environmental Risk Factor Etiology Event Family Fluorescence Microscopy Functional disorder Future Gene Expression Profiling General Population Generations Genes Genetic Genetic Predisposition to Disease Genetic Transcription Genetic study Genomic approach Goals Health Hemorrhage Heterozygote Histologic Homeostasis Human Individual Intracranial Aneurysm Intracranial Hemorrhages Knock-out Left Light Long-Term Care Mediating Medicine Modeling Molecular Morbidity - disease rate Morphology Mus Mutation Nature Neurologic Deficit Organoids Outcome Pathogenesis Pathogenicity Pathologic Pathway interactions Patients Phenotype Predisposition Prevalence Proteins Public Health Reporter Reporting Research Risk Risk Factors Role Rupture Ruptured Aneurysm Series Stroke Structure Subarachnoid Hemorrhage Survivors Testing Therapeutic Therapeutic Intervention Transgenic Organisms Variant Visualization WNT Signaling Pathway Work Zebrafish blood vessel development cerebral artery cerebrovascular cohort exome sequencing experimental study functional genomics genetic risk factor genome editing genome wide association study hemodynamics improved in vivo loss of function member microCT model organism mortality mouse model mutant novel precision medicine risk variant senescence targeted treatment transcriptional reprogramming

项目摘要

ABSTRACT Saccular intracranial aneurysms (IA) represent a significant health issue in the US and worldwide. IA rupture leads to intracranial hemorrhage, with devastating outcomes: 30% of patients die within a month of the initial event, and 50% of survivors are left with severe neurological deficits requiring long-term care. IA is a multi- factorial disorder underlain by genetic and environmental risk factors. In a series of genome-wide association studies we identified multiple common variants that contribute to IA risk. To discover rare coding variants with large-effect size, we conducted whole exome sequencing analysis of a cohort of >250 patients from 58 families, each with at least 3 members diagnosed with IA. This analysis identified heterozygous rare and deleterious mutations in two novel genes, WBP11 and PPIL4, which collectively explain more than 10% of familial IA cases in our cohort. In preliminary experiments, we demonstrated that loss- of-function ppil4 and wbp11 zebrafish and mouse models display dramatic and similar changes in cerebrovascular morphology and cerebral hemorrhage, suggesting convergent action, and uncovered a specific requirement for both genes in endothelial cells (ECs). We also determined that depletion of WBP11 or PPIL4 in human ECs induces a pathological transcriptional reprogramming towards a senescent cellular state. Collectively, these findings led to the hypotheses that a) PPIL4 and WBP11 are critical components of a network that regulates cerebrovascular morphology and EC homeostasis; b) pathological EC reprogramming is the underlying mechanism in IA associated PPIL4 and WBP11 variants; and c) PPIL4 and WBP11 convergent action to the Wnt signaling pathway is mediated through binding partners and disrupted by IA-associated mutations. In this proposal we apply experimental and functional genomics approaches to test these hypotheses, aiming to establish the converging role of WBP11 and PPIL4 in cerebrovascular ECs, following the discovery of rare- deleterious coding mutations associated with familial IA. The expected outcome of this work is to define and integrate multiple facets that underpin the function of WBP11 and PPIL4: 1) the morphologic and histologic consequences of global and EC-specific deficiency of WBP11 and PPIL4 for structural integrity of the cerebral vessels in model organisms; 2) the impact of WBP11 and PPIL4 depletion and IA-associated variants on EC function at the cellular and molecular level; and 3) the convergence of WBP11 and PPIL4 in ECs via activation of Wnt signaling. These outcomes will inform a framework implicating pathological EC reprogramming as an overlooked contributing factor in IA etiology, while providing a novel conceptual framework for IA pathophysiology, with the long-term goal to develop precision medicine strategies to improve diagnosis and molecularly informed therapeutic interventions for IA patients.

抽象的 accular颅内动脉瘤（IA）代表了美国和全球的重大健康问题。 IA破裂导致颅内出血，造成毁灭性的结果：30％的患者在初始的一个月内死亡事件和50％的幸存者留下了需要长期护理的严重神经缺陷。 ia是一个多遗传和环境危险因素是阶乘障碍。在一系列全基因组关联中研究我们确定了导致IA风险的多种常见变体。为了发现具有较大效果大小的稀有编码变体，我们对来自58个家庭的> 250例患者组成的队列，每个患者至少有3名被诊断为IA的成员。这个分析在两个新型基因WBP11和PPIL4中鉴定出杂合的稀有和有害突变，它们统称为在我们的队列中解释超过10％的家族IA病例。在初步实验中，我们证明了损失 - 功能PPIL4和WBP11斑马鱼和鼠标模型显示出巨大的变化脑血管形态和脑出血，提示收敛作用，并发现了特定的内皮细胞（EC）中这两个基因的需求。我们还确定WBP11或PPIL4在人类ECS诱导了对衰老细胞状态的病理转录重编程。总的来说，这些发现导致了a）ppil4和wbp11是网络的关键组成部分的假设调节脑血管形态和EC稳态； b）病理EC重编程是 IA相关的PPIL4和WBP11变体中的基本机制； c）PPIL4和WBP11收敛动作 Wnt信号通路是通过结合伙伴介导的，并被IA相关的突变扰动。在此提案中，我们采用实验和功能基因组学方法来检验这些假设，旨在在发现罕见的情况下，建立WBP11和PPIL4在脑血管EC中的融合作用与家族性IA相关的有害编码突变。这项工作的预期结果是定义和整合了WBP11和PPIL4的功能的多个方面：1）形态学和组织学 WBP11和PPIL4全球和EC特异性缺乏对大脑结构完整性的后果模型生物中的血管； 2）WBP11和PPIL4耗竭以及与IA相关的变体对EC的影响细胞和分子水平的功能； 3）通过激活在EC中WBP11和PPIL4的收敛性 Wnt信号传导。这些结果将为涉及病理EC重编程的框架提供信息忽视了IA病因的贡献因素，同时为IA提供了一种新颖的概念框架病理生理学，其长期目标是制定精确的医学策略以改善诊断和 IA患者的分子知情治疗干预措施。