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Identifying the role of notch3 in brain pericyte function in health and Alzheimer's disease

确定 notch3 在健康和阿尔茨海默病中大脑周细胞功能中的作用

基本信息

批准号：
10679198
负责人：
Richard Daneman
金额：
$ 183.88万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
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：
2023-04-15 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10679198
关键词：
Adult Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease patient Astrocytes Automobile Driving Biological Assay Blood Blood - brain barrier anatomy Blood Circulation Blood brain barrier dysfunction Brain Brain Diseases CNS degeneration CRISPR interference CRISPR-mediated transcriptional activation CRISPR/Cas technology Cell Communication Cell Line Cells Central Nervous System ChIP-seq Coculture Techniques Data Development Disease Endothelial Cells Endothelium Epilepsy Extracellular Matrix Functional disorder Gene Expression Profile Generations Genes Genetic Genetic Transcription Genomic approach Health Human Immune Impaired cognition Knockout Mice Knowledge Laboratories Leukocyte Adhesion Molecules Link Maintenance Mediating Mesoderm Modeling Molecular Multiple Sclerosis NOTCH3 gene Neural Crest Neurons Organ Pathogenesis Pathology Pathway interactions Patients Pericytes Permeability Phenotype Pluripotent Stem Cells Process Property Prosencephalon Protocols documentation Regulation Role Serum Signal Transduction Smooth Muscle Myocytes Somatic Cell Specific qualifier value Stroke Supporting Cell System Tight Junctions Transport Process Traumatic Brain Injury Up-Regulation Vascular Smooth Muscle blood-brain barrier function brain endothelial cell cell type extracellular facsimile fetal human disease human model human pluripotent stem cell improved in vitro Model in vivo innovation insight mouse genetics mouse model nervous system disorder neurovascular unit notch protein progenitor programs restoration self assembly stem cell technology transcription factor transcriptome transcytosis

项目摘要

ABSTRACT The BBB acts as a signaling and transport interface between the blood and brain, and with its very low permeability and a wealth of molecular transport systems, the BBB helps regulate the extracellular composition of the brain. While brain microvascular endothelial cells (BMECs) are possessive of these BBB functions, the BBB is greatly influenced by interactions with supporting cells of the neurovascular unit (NVU) such as astrocytes, pericytes and neurons. Recent studies have indicated the importance of CNS pericytes in BBB formation and maintenance, with pericytes triggering reduced transcytosis, reduced expression of leukocyte adhesion molecules and proper tight junction organization in BMECs. Loss of pericyte-endothelial cell interactions and BBB dysfunction are thought to be critical for the pathogenesis of Alzheimer’s disease. Despite the potential importance, the molecular mechanisms driving brain pericyte regulation of the BBB in health and Alzheimer’s disease are largely unknown, particularly in humans. In this proposal, we aim to further examine the mechanisms by which brain pericytes are specified and subsequently impact BBB function. A powerful and innovative approach to explore human pericyte development and function is the use of human pluripotent stem cell (hPSC) technology to model brain pericytes. However, current strategies for differentiating brain pericytes result in cells that lack key brain pericyte hallmarks. We have devised a protocol where brain pericytes can be differentiated from hPSCs by activation of Notch3 signaling in neural crest, yielding improved facsimiles of in vivo brain pericytes. Here, we will further explore the impact of Notch3 signaling on pericyte development and the subsequent effects on BBB induction and maintenance in health and disease. The impact of Notch3 activation in hPSC-derived neural crest will be evaluated by assessing brain pericyte fate, pericyte functionality and the ability to induce BBB properties in co-cultured BMECs. In parallel, using genomics approaches, we have identified a transcriptional network directly regulated by Notch3 activation in pericytes, and have found that this network is downregulated in brain pericytes of Alzheimer’s disease patients. To elucidate the mechanism by which Notch3 activation drives pericyte specification and development, we will use complementary tools of CRISPR-edited hPSC lines and developmental mouse models to systematically regulate the Notch3 transcriptional network and determine impacts on pericyte development and BBB formation and maintenance. Finally, we will assess whether upregulation of Notch signaling in a mouse model of Alzheimer’s disease can ameliorate the effects of pericyte dysfunction on BBB pathophysiology associated with Alzheimer’s disease. Taken together, understanding the impact of Notch3 signaling on pericyte development and BBB function could yield many new mechanistic insights about human BBB induction and maintenance and open new avenues for restoring BBB function in Alzheimer’s disease.

摘要血脑屏障作为血液和大脑之间的信号传导和运输界面，其非常低的血脑屏障水平，血脑屏障具有良好的通透性和丰富的分子转运系统，有助于调节细胞外成分大脑。虽然脑微血管内皮细胞（BMEC）具有这些BBB功能，但脑微血管内皮细胞（BMEC）的功能可能与BBB的功能有关。 BBB受到与神经血管单位（NVU）的支持细胞的相互作用的极大影响，星形胶质细胞、周细胞和神经元。近年来的研究表明，中枢神经系统周细胞在血脑屏障中的重要性形成和维持，周细胞触发减少的转胞吞作用，减少的白细胞表达 BMEC中的粘附分子和适当的紧密连接组织。周细胞-内皮细胞丢失相互作用和BBB功能障碍被认为是阿尔茨海默病发病机制的关键。尽管有潜在的重要性，但在脑缺血中驱动脑周细胞调节BBB的分子机制仍不清楚。健康和阿尔茨海默病在很大程度上是未知的，特别是在人类中。在这项建议中，我们的目标是进一步研究脑周细胞被指定并随后影响BBB功能的机制。一一个强大的和创新的方法来探索人类周细胞的发育和功能是利用人类多能干细胞（hPSC）技术来模拟脑周细胞。然而，目前的差异化战略脑周细胞导致缺乏关键脑周细胞标志的细胞。我们设计了一个方案周细胞可以通过激活神经嵴中的Notch 3信号传导而从hPSC分化，产生改善的细胞分化。活体脑周细胞的复制品。在这里，我们将进一步探讨Notch 3信号对周细胞的影响，发展和随后对健康和疾病中BBB诱导和维持的影响。的 Notch 3活化在hPSC衍生神经嵴中的影响将通过评估脑周细胞命运，周细胞功能和诱导共培养的BMEC中的BBB性质的能力。同时，使用通过基因组学方法，我们已经确定了一个直接受Notch 3激活调控的转录网络，周细胞，并发现该网络在阿尔茨海默病患者的脑周细胞中下调。为了阐明Notch 3激活驱动周细胞特化和发育的机制，我们将使用CRISPR编辑的hPSC系和发育小鼠模型的互补工具，调节Notch 3转录网络并确定对周细胞发育和BBB的影响形成和维持。最后，我们将评估是否上调Notch信号在小鼠模型中，可以改善周细胞功能障碍对BBB病理生理学的影响，患有老年痴呆症总之，了解Notch 3信号对周细胞的影响发展和血脑屏障功能可以产生许多新的机制的见解，人类血脑屏障诱导和维持并为恢复阿尔茨海默病中的BBB功能开辟新途径。