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.

抽象的囊状颅内动脉瘤（IA）是美国和全世界的一个重大健康问题。内膜破裂导致颅内出血，造成毁灭性后果：30% 的患者在初次治疗后一个月内死亡事件中，50% 的幸存者患有严重的神经功能缺损，需要长期护理。 IA 是一个多由遗传和环境危险因素引起的因子紊乱。在一系列全基因组关联中在研究中，我们发现了多种导致 IA 风险的常见变异。为了发现具有大效应量的罕见编码变异，我们进行了全外显子组测序分析来自 58 个家庭的超过 250 名患者组成的队列，每个家庭至少有 3 名成员被诊断患有 IA。本次分析在两个新基因 WBP11 和 PPIL4 中发现了杂合的罕见和有害突变，这两个基因共同解释了我们队列中超过 10% 的家族性 IA 病例。在初步实验中，我们证明了损失功能丧失的 ppil4 和 wbp11 斑马鱼和小鼠模型在脑血管形态和脑出血，提示收敛作用，并发现了一个特定的内皮细胞（EC）中对这两个基因的需求。我们还确定 WBP11 或 PPIL4 的耗尽人类内皮细胞诱导病理性转录重编程，导致细胞衰老状态。总的来说，这些发现得出了以下假设：a) PPIL4 和 WBP11 是网络的关键组成部分调节脑血管形态和 EC 稳态； b) 病理性 EC 重编程是 IA 相关 PPIL4 和 WBP11 变体的潜在机制； c) PPIL4 和 WBP11 趋同作用 Wnt 信号通路通过结合伴侣介导，并被 IA 相关突变破坏。在本提案中，我们应用实验和功能基因组学方法来测试这些假设，旨在继发现罕见的- 与家族性 IA 相关的有害编码突变。这项工作的预期成果是定义和整合支持 WBP11 和 PPIL4 功能的多个方面：1) 形态学和组织学 WBP11 和 PPIL4 的整体和 EC 特异性缺陷对大脑结构完整性的影响模式生物中的血管； 2) WBP11和PPIL4耗竭以及IA相关变异对EC的影响在细胞和分子水平上发挥作用； 3) WBP11 和 PPIL4 通过激活在 EC 中融合 Wnt 信号传导。这些结果将为一个框架提供信息，该框架将病理性 EC 重编程作为一种忽视了 IA 病因学的影响因素，同时为 IA 提供了一个新颖的概念框架病理生理学，长期目标是开发精准医学策略以改善诊断和治疗 IA 患者的分子信息治疗干预。