CCN1-induced Senescence in the Cerebral Vasculature

CCN1 诱导的脑血管衰老

基本信息

批准号：
10370877
负责人：
Jose Felix Moruno Manchon
金额：
$ 23.4万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10370877
关键词：
Aging Bilateral Binding Binding Proteins Blood - brain barrier anatomy Blood Vessels Brain Carotid Stenosis Cell Aging Cell Cycle Cell Cycle Progression Cell Proliferation Cell physiology Cells Cerebrovascular Circulation Cerebrovascular system Cerebrum Chronic Cognitive Cognitive deficits Coupled Cysteine DNA DNA Damage Data Dementia Disease Down-Regulation Endothelial Cells Endothelium Extracellular Matrix Proteins FMR1 Fibroblasts Functional disorder G-Quartets GTP-Binding Protein alpha Subunits, Gs Gait Genes Guanine Heterochromatin Impaired cognition Impairment Integrin alpha6beta1 Integrins Ligands Link Malignant Neoplasms Measures Mediating Messenger RNA Microscopy Modeling Molecular Motor Mus Operative Surgical Procedures Pathologic Patients Permeability Phenotype Play Protein Deficiency Proteins RNA RNA-Binding Proteins Ribosomes Role Scheme Secondary to Signal Pathway Structure System Techniques Testing Tissues Translations Tumor Suppressor Proteins Up-Regulation Viral Vector adhesion receptor aged aging brain blood-brain barrier disruption brain endothelial cell cell injury cerebral hypoperfusion cerebrovascular pathology cognitive function conditioned fear cysteine rich protein dementia risk design motor deficit mouse model mutant negative affect novel object recognition overexpression prevent senescence vascular cognitive impairment and dementia vector white matter

项目摘要

Project Summary Vascular contributions to cognitive impairment and dementia (VCID) includes a number of different disorders identified by cognitive deficits secondary to cerebrovascular pathology. Senescence in the cerebral endothelium has been proposed as a mechanism that can drive blood brain barrier disruption (BBB), which precedes VCID. However, the mechanisms that drive endothelial cells to senescence are not fully understood. Guanine-quadruplex (G4) is a non-canonical secondary structure formed in G-rich sequences in DNA and RNA. Importantly, G4 stabilization promotes cellular senescence. Particularly, stabilization of G4s in mRNA (rG4s) stalls ribosomes in the elongation stage and blocks translation. This mechanism is promoted by rG4- binding proteins. Fragile X mental retardation protein (FMRP) binds to the rG4 of tumor suppressors and components of signaling pathways involved in senescence. FMRP deficiency has been associated with senescence in different tissues. We found that overexpressing FMRP in cultured cerebral endothelial cells (CEC) mitigated DNA damage, an important cause of cell senescence. We found that G4 stabilization promoted endothelial senescence and enhanced the levels of Cysteine-rich angiogenic inducer 61 (Cyr61/CCN1), a matricellular protein secreted by endothelial cells in the brain. CCN1 is considered a component of the senescence-associated secretory phenotype (SASP). In fibroblasts, CCN1 induces senescence by binding to integrin α6β1. In our study, we will use bilateral carotid artery stenosis (BCAS) that induces chronic cerebral hypoperfusion and models VCID in mice. We found that BCAS mice showed enhanced levels of G4s, CCN1 and senescence markers. We hypothesize that the endothelial senescence associated with G4 stabilization could be mitigated in mice and hence prevent BBB disruption and cognitive deficits. To test our hypothesis, we propose two aims: In Aim 1, we will selectively target the brain endothelium of young mice with a vector encoding a mutant form of CCN1 that cannot bind to α6β1. We will measure motor and cognitive function, and BBB permeability, and senescence in the brain. In Aim 2, we will determine if upregulating FMRP mitigates motor and cognitive deficits, BBB impairment, and senescence in the cerebral vasculature of aged BCAS mice. Overall, this study proposes two independent strategies to mitigate VCID-associated phenotypes induced by senescence in the cerebrovasculature: impeding the CCN1-α6β1binding in the cerebral endothelium; and upregulating FMPR specifically in the brain vasculature of mice.

项目摘要对认知障碍和痴呆（VCID）的血管贡献包括许多不同的疾病通过继发于脑血管病理的认知缺陷鉴定。大脑内皮衰老已提出是一种可能驱动血脑屏障破坏（BBB）的机制，该机制在VCID之前。但是，尚未完全了解将内皮细胞驱动到感应的机制。鸟嘌呤 - 四链体（G4）是一种非典型的二级结构，在DNA和 RNA。重要的是，G4稳定促进了细胞感应。特别是在mRNA中G4S的稳定（RG4）在伸长阶段失速核糖体并阻止翻译。这种机制是由RG4-促进的结合蛋白。脆弱的X智力低下蛋白（FMRP）与肿瘤补充剂的RG4结合和信号通路的组成部分涉及感应。 FMRP缺乏与在不同组织中的感应。我们发现在培养的脑内皮细胞中过表达FMRP（CEC）减轻DNA损伤，这是细胞感应的重要原因。我们发现G4稳定促进了内皮感应并增强了富含半胱氨酸的水平血管生成诱导剂61（CYR61/CCN1），一种由大脑内皮细胞分泌的基质蛋白。 CCN1是被认为是相关的秘书表型（SASP）的组成部分。在成纤维细胞中，CCN1 通过与整联蛋白α6β1结合来诱导衰老。在我们的研究中，我们将使用双侧颈动脉狭窄（BCA），诱导慢性脑部灌注和小鼠模型VCID。我们发现BCA小鼠的G4S，CCN1和衰老水平增强标记。我们假设在小鼠和因此，防止BBB破坏和认知缺陷。为了检验我们的假设，我们提出了两个目标：在AIM 1中，我们将使用编码突变体形式的向量有选择地靶向幼鼠的脑内皮无法与α6β1结合的CCN1。我们将测量运动和认知功能，以及BBB的渗透性，以及大脑衰老。在AIM 2中，我们将确定上调FMRP是否减轻运动和认知定义，BBB损害和老年BCA小鼠的脑脉管系统中的衰老。总体而言，这项研究提出了两种独立策略，以减轻由VCID相关的表型引起的脑血管系统中的敏感性：阻碍脑内皮中的CCN1-α6β1BIND；和在小鼠的脑脉管系统中特别上调FMPR。