Selective neurovascular regulation by a vascular dementia-related noncoding RNA Snord118

血管性痴呆相关非编码 RNA Snord118 的选择性神经血管调节

基本信息

批准号：
10435866
负责人：
Jianfu Chen
金额：
$ 46.75万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
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：
2022-05-15 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10435866
关键词：
Address Affect Alzheimer&apos s Disease Alzheimer&apos s disease related dementia Astrocytes Base Pairing Binding Biogenesis Biological Assay Biology Blood - brain barrier anatomy Blood Vessels Brain Cell Line Cell model Cell physiology Cells Cerebral hemisphere hemorrhage Cerebrum Code Cyst Data Defect Detection Disease Electrical Resistance Endothelial Cells Event Functional disorder Generations Genes Goals Human Impairment Induced Mutation Knock-in Mouse Lesion Leukoencephalopathy Maps Mediating Messenger RNA Methods Microglia Microvascular Dysfunction Molecular Target Morphology Motor Mutation Names Neurodegenerative Disorders Neuroglia Neurologic Neurons Oligodendroglia Organ Outcome Paris, France Pathogenesis Pathologic Patients Pericytes Permeability Phenotype Point Mutation Process Property Proteins Protocols documentation Psoralens RNA RNA analysis RNA methylation RNA, Ribosomal, 28S Regulation Resolution Ribosomal Proteins Ribosomal RNA Ribosomes Small Nucleolar RNA Smooth Muscle Myocytes Specificity Stem Cell Research Structure Structure-Activity Relationship Technology Testing Tight Junctions Tissues Tube Untranslated RNA Vascular Dementia Vascular Smooth Muscle base blood-brain barrier permeabilization brain endothelial cell calcification cell type cognitive function crosslink data tools genome-wide improved induced pluripotent stem cell mutant nervous system disorder neurovascular neurovascular unit novel single molecule transcytosis white matter

Address Affect Alzheimer&apos s Disease Alzheimer&apos s disease related dementia Astrocytes Base Pairing Binding Biogenesis Biological Assay Biology Blood - brain barrier anatomy Blood Vessels Brain Cell Line Cell model Cell physiology Cells Cerebral hemisphere hemorrhage Cerebrum Code Cyst Data Defect Detection Disease Electrical Resistance Endothelial Cells Event Functional disorder Generations Genes Goals Human Impairment Induced Mutation Knock-in Mouse Lesion Leukoencephalopathy Maps Mediating Messenger RNA Methods Microglia Microvascular Dysfunction Molecular Target Morphology Motor Mutation Names Neurodegenerative Disorders Neuroglia Neurologic Neurons Oligodendroglia Organ Outcome Paris, France Pathogenesis Pathologic Patients Pericytes Permeability Phenotype Point Mutation Process Property Proteins Protocols documentation Psoralens RNA RNA analysis RNA methylation RNA, Ribosomal, 28S Regulation Resolution Ribosomal Proteins Ribosomal RNA Ribosomes Small Nucleolar RNA Smooth Muscle Myocytes Specificity Stem Cell Research Structure Structure-Activity Relationship Technology Testing Tight Junctions Tissues Tube Untranslated RNA Vascular Dementia Vascular Smooth Muscle base blood-brain barrier permeabilization brain endothelial cell calcification cell type cognitive function crosslink data tools genome-wide improved induced pluripotent stem cell mutant nervous system disorder neurovascular neurovascular unit novel single molecule transcytosis white matter

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项目摘要

PROJECT SUMMARY/ABSTRACT Dysfunction of neurovascular unit (NVU) is a key pathological event of neurodegenerative diseases, including Alzheimer's disease and Alzheimer's disease-related dementias (AD/ADRD). The mechanism underlying cell- type selective vulnerability in NVU is poorly understood. The goal of this proposal is to establish a novel mechanism by which NVU cell(s) are selectively impaired by the disruption of a global ribosome biogenesis. We will focus on a newly identified ribosomopathy disease gene Snord118, which encodes a noncoding RNA acting as a ribosome biogenesis factor. This is interesting because Snord118 mutations cause the first purely neurological disorder in ribosomopathies, named leukoencephalopathy with calcifications and cysts (LCC), with NVU lesions. There is very little understanding of Snord118 and LCC pathogenesis. This study has an opportunity to determine functions and mechanisms of Snord118 and LCC disease. We have assembled the following preliminary data: 1) generated two disease point mutation knock-in (KI) mice, which display early pericyte and BBB defects. These results suggest that brain endothelial cells (ECs) and pericytes are selectively affected in LCC, which justifies our iPSC research focus on brain ECs and pericytes; 2) generated five Snord118 mutant iPSC lines with isogenic controls, established protocols of directing iPSCs into brain microvascular endothelial cells (BMECs) and pericytes with the CNS identities, and prepared functional assays for BMEC, pericyte, and blood-brain barrier (BBB) properties; 3) developed the PARIS method to high throughput map RNA structures and identify RNA targets at single molecule and genome-wide levels with base-pair resolution. Our PARIS revealed a dynamic RNA structure and interaction network in Snord118 ribosome biogenesis and LCC. Leveraging on these preliminary data and tools, we propose to test the hypothesis that Snord118 mutation-mediated disruption of ribosome biogenesis selectively affects BMECs and pericytes via targeting rRNAs and non-rRNAs. Aim 1 will determine cellular functions of Snord118 in neurovascular cells focusing on BMECs and pericytes. Aim 2 will identify Snord118 targets and its RNA structure-function relationships. Overall, using our new iPSC-derived NVU cells and latest PARIS2, this study will generate the first human cellular models that do not currently exist for SNORD118 LCC, identify mechanisms of SNORD118 action and LCC disease, uncover a previously unknown vulnerability of specific NVU cells to the disruption of a ubiquitous ribosome biogenesis process, and therefore help to reconcile the neurological phenotype specificity of ribosomopathies with the global requirement for ribosome biogenesis.

项目摘要/摘要神经血管单元（NVU）功能障碍是神经退行性疾病的关键病理事件，包括阿尔茨海默氏病和阿尔茨海默氏病与疾病有关的痴呆症（AD/ADRD）。细胞的基础机制 NVU中的选择性脆弱性知之甚少。该提议的目的是建立小说 NVU细胞被整体核糖体生物发生的破坏选择性损害的机制。我们将重点关注新鉴定的核糖瘤病疾病基因SNORD118，该疾病基因SNORD118编码非编码RNA 充当核糖体生物发生因子。这很有趣，因为snord118突变导致第一个纯粹导致核糖症的神经系统疾病，称为钙化和囊肿（LCC）的白斑脑病（LCC），具有 NVU病变。对SNORD118和LCC发病机理的了解很少。这项研究有一个确定SNORD118和LCC病的功能和机制的机会。我们组装了遵循初步数据：1）产生两种疾病点突变敲击（Ki）小鼠，它们早期显示周细胞和BBB缺陷。这些结果表明脑内皮细胞（EC）和周细胞有选择性在LCC中受到影响，这证明了我们的IPSC研究专注于脑EC和周细胞； 2）产生五个 SNORD118具有同基因控制的突变体IPSC线，已建立的将IPSC引导到大脑的方案微血管内皮细胞（BMEC）和具有CNS身份的周细胞，并制备的功能测定对于BMEC，周细胞和血脑屏障（BBB）特性； 3）将巴黎方法开发为高吞吐量图RNA结构并在单分子和全基因组水平上鉴定RNA靶标碱基对分辨率。我们的巴黎揭示了SNORD118中的动态RNA结构和相互作用网络核糖体生物发生和LCC。利用这些初步数据和工具，我们建议测试 SNORD118突变介导的核糖体生物发生的破坏选择性影响BMEC和通过靶向RRNA和非RRNA的周细胞。 AIM 1将确定SNORD118的细胞功能致力于BMEC和周细胞的神经血管细胞。 AIM 2将识别SNORD118目标及其RNA 结构功能关系。总体而言，使用我们新的IPSC衍生的NVU细胞和最新的Paris2，这项研究将生成当前为SNORD118 LCC不存在的第一个人类细胞模型，识别 SNORD118动作和LCC疾病的机制，揭示了以前未知的特定脆弱性 NVU细胞破坏无处不在的核糖体生物发生过程，因此有助于调和核糖体病的神经表型特异性具有核糖体生物发生的全球需求。