Endothelial Cell Reprogramming in Familial Intracranial Aneurysm

家族性颅内动脉瘤的内皮细胞重编程

• 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
• 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

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.

摘要 囊状颅内动脉瘤（IA）是美国和全球的一个重大健康问题。IA破裂 会导致颅内出血，并带来毁灭性的后果：30%的患者在最初的一个月内死亡。 50%的幸存者留下了严重的神经功能缺损，需要长期护理。它是一个多- 由遗传和环境危险因素引起的因素性疾病。在一系列的基因组关联中 研究中，我们发现了多种常见的变异，有助于IA的风险。 为了发现具有大效应大小的罕见编码变体，我们对 来自58个家庭的>250名患者的队列，每个家庭至少有3名成员被诊断患有IA。该分析 在两个新基因WBP 11和PPIL 4中鉴定了杂合的罕见和有害突变， 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- 功能缺失的ppil 4和wbp 11斑马鱼和小鼠模型显示出显著的和相似的变化， 脑血管形态和脑出血，表明收敛作用，并揭示了一个特定的 这两种基因在内皮细胞（EC）中的需求。我们还确定了在细胞中WBP 11或PPIL 4的缺失， 人EC诱导病理性转录重编程朝向衰老细胞状态。 总的来说，这些发现导致了以下假设：a）PPIL 4和WBP 11是网络的关键组成部分 B）病理性EC重编程是脑血管形态学和EC稳态的调节因子; IA相关PPIL 4和WBP 11变体的潜在机制;和c）PPIL 4和WBP 11会聚作用 与Wnt信号通路的相互作用是通过结合伴侣介导的，并被IA相关突变破坏。 在这项提议中，我们应用实验和功能基因组学方法来验证这些假设， 在发现罕见的- 与家族性IA相关的有害编码突变。这项工作的预期成果是界定和 整合支持WBP 11和PPIL 4功能的多个方面：1）形态学和组织学 WBP 11和PPIL 4的整体和EC特异性缺陷对大脑结构完整性的影响 2）WBP 11和PPIL 4缺失以及IA相关变体对EC的影响 WBP 11和PPIL 4在内皮细胞中的会聚是通过激活 Wnt信号这些结果将为涉及病理性EC重编程的框架提供信息， 在IA病因学中被忽视的因素，同时为IA提供了一个新的概念框架 病理生理学，长期目标是制定精准医学策略，以改善诊断和 为IA患者提供分子信息的治疗干预。