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

• 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
• 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' s Disease; Alzheimer' 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

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.

项目总结/摘要 神经血管单位（NVU）功能障碍是神经退行性疾病的关键病理事件，包括 阿尔茨海默病和阿尔茨海默病相关痴呆（AD/ADRD）。细胞的潜在机制- NVU中的类型选择性漏洞知之甚少。这项提案的目的是建立一个新的 NVU细胞通过全局核糖体生物合成的破坏而选择性受损的机制。 我们将重点关注一个新发现的核糖体病基因Snord 118，它编码一个非编码RNA， 作为核糖体生物合成因子。这很有趣，因为Snord 118突变导致了第一个纯粹的 核糖体病中的神经系统疾病，称为伴有钙化和囊肿的白质脑病（LCC）， NVU病变。Snord 118和LCC发病机制的了解很少。这项研究有一个 有机会确定Snord 118和LCC疾病的功能和机制。我们已经聚集了 以下初步数据：1）产生了两个疾病点突变敲入（KI）小鼠，其显示早期 周细胞和血脑屏障缺损。这些结果表明，脑内皮细胞（ECs）和周细胞选择性地 在LCC中受到影响，这证明了我们的iPSC研究重点是脑EC和周细胞; 2）产生了五个 Snord 118突变iPSC系与同基因对照，建立了将iPSC导入脑的方案 微血管内皮细胞（BMEC）和周细胞与中枢神经系统的身份，并准备功能测定 BMEC，周细胞和血脑屏障（BBB）特性; 3）开发了巴黎方法，以提高 在单分子和全基因组水平上， 碱基对分辨率我们的巴黎揭示了Snord 118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 Snord 118突变介导的核糖体生物发生的破坏选择性地影响BMEC和 通过靶向rRNA和非rRNA来抑制周细胞。目的1将确定Snord 118在细胞中的功能。 神经血管细胞集中于BMEC和周细胞。目标2将识别Snord 118靶标及其RNA 结构-功能关系。总的来说，使用我们新的iPSC衍生的NVU细胞和最新的PARIS 2，这项研究 将为SNORD 118 LCC生成目前尚不存在的第一个人类细胞模型， SNORD 118作用和LCC疾病的机制，揭示了一个以前未知的特定的脆弱性， NVU细胞破坏一个普遍存在的核糖体生物合成过程，因此有助于调和 核糖体病的神经系统表型特异性与核糖体生物合成的全球需求。