The roles of pericyte-derived laminin in neurovascular function and neurodegeneration

周细胞源性层粘连蛋白在神经血管功能和神经变性中的作用

• 批准号: 10296497
• 负责人: Yao Yao
• 金额: $ 181.11万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10296497
• 关键词: Affect; Age; Alzheimer' s Disease; Alzheimer' s disease related dementia; Amyloid beta-Protein; Astrocytes; Basement membrane; Biochemical; Biological Assay; Biological Process; Blood - brain barrier anatomy; Blood Vessels; Brain; Cells; Cerebrovascular Circulation; Defect; Deposition; Deterioration; Development; Disease; Endothelial Cells; Extracellular Matrix; Functional disorder; Generations; Image; Impaired cognition; Impairment; In Vitro; Knockout Mice; Label; Laboratories; Laminin; Lead; Maintenance; Mediating; Mission; Molecular; Molecular Target; Molecular Weight; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neuronal Injury; Neurons; Pathogenesis; Perfusion; Pericytes; Protein Isoforms; Proteins; Quantitative Autoradiography; Research; Role; Smooth Muscle Myocytes; Structure; Testing; Therapeutic; Thick; Vascular Smooth Muscle; age group; age related; blood-brain barrier disruption; cognitive disability; conditional knockout; effective therapy; fluorescein isothiocyanate dextran; in vivo; innovation; laminin S; macromolecule; mouse model; neuron loss; neuronal survival; neurovascular; neurovascular unit; novel; receptor; two photon microscopy; two-photon

Project Summary/Abstract The long-term objective of this application is to understand whether the basement membrane (BM), the non-cellular component of the neurovascular unit, is involved in the pathogenesis of Alzheimer’s disease and can be targeted to treat Alzheimer’s disease and Alzheimer’s disease-related dementias. This is consistent with the mission of NIA. This proposal aims to investigate the biological functions of pericytic laminin in: (1) neurovascular function, including blood brain barrier (BBB) integrity, cerebral blood flow (CBF) and brain influx/efflux function, and (2) neuronal survival/function. In Aim 1, the function of pericytic laminin in BBB integrity will be investigated. First, whether and to what extent loss of pericytic laminin affects BBB integrity will be investigated using FITC-Dextrans of various molecular weights (Aim 1A). Next, the molecular mechanism underlying loss of pericytic laminin-induced BBB breakdown will be investigated, with a focus on changes in paracellular and transcellular transport in endothelial cells (Aim 1B). Furthermore, the receptors that mediate pericytic laminin’s effect in endothelial cells will be identified and examined (Aim 1C). In Aim 2, the function of pericytic laminin in CBF will be investigated. First, how loss of pericytic laminin affects CBF will be investigated using quantitative autoradiography and two-photon imaging (Aim 2A). Next, whether the reduced CBF is caused by pericyte loss/degeneration will be investigated in vitro and in vivo (Aim 2B). Furthermore, the receptors that mediate pericytic laminin’s effect in pericytes will be identified and examined (Aim 2C). In Aim 3, the role of pericytic laminin in brain influx/efflux function will be investigated. First, whether and how loss of pericytic laminin affects brain influx/efflux function will be investigated by influx/efflux assays using various fluorescently labeled macromolecules (Aim 3A). Next, whether the impaired influx/efflux function is due to BM damage and how loss of pericytic laminin affects BM composition/structure will be explored (Aim 3B). In Aim 4, the function of pericytic laminin in neuronal injury/neurodegeneration will be investigated. In this aim, whether loss of pericytic laminin leads to neuronal injury/neurodegeneration will be investigated at biochemical, structural, and functional levels. In addition, the age at which neuronal injury/neurodegeneration occurs will be determined and compared to that at which neurovascular dysfunction occurs. Successful completion of this study will elucidate the fundamental roles of pericytic laminin in neurovascular function and neuronal survival/function, and identify novel molecular targets with therapeutic potential in Alzheimer’s disease and Alzheimer’s disease-related dementias. In addition, this proposal may also lead to the generation of an innovative mouse model for neurodegeneration and open doors for new research.

项目摘要/摘要 本应用的长期目的是了解地下室膜（BM）是否是 神经血管单元的非细胞成分参与阿尔茨海默氏症的发病机理 疾病可以针对治疗阿尔茨海默氏病和阿尔茨海默氏病与疾病相关的痴呆症。 这与NIA的使命是一致的。该建议旨在调查 周围层粘连蛋白在：（1）神经血管功能，包括血脑屏障（BBB）完整性，脑障碍 血流（CBF）和大脑影响/外排功能，以及（2）神经元存活/功能。在AIM 1中 将研究周围层粘连蛋白在BBB完整性中的功能。首先，无论损失何种程度 将使用各种分子的FITC-脱发射研究，将研究周围层粘连蛋白的影响BBB的完整性 权重（AIM 1A）。接下来，细胞周层粘连蛋白诱导的BBB的分子机制 将研究崩溃，重点是细胞细胞和跨细胞运输的变化 内皮细胞（AIM 1B）。此外，介导周细胞层粘连蛋白作用的接收器 将识别和检查内皮细胞（AIM 1C）。在AIM 2中，周细胞层粘连蛋白的功能 CBF将被调查。首先，将如何使用周细层粘连蛋白影响CBF的损失。 定量放射自显影和两光子成像（AIM 2A）。接下来，是否减少了CBF 由周细胞丧失/变性引起的体外和体内将研究（AIM 2B）。此外， 将鉴定并检查介导周细胞周围层粘蛋白作用的受体（AIM 2C）。 AIM 3，将研究周细层粘连蛋白在大脑影响/外排功能中的作用。首先，是否和 影响/外排的损失如何影响脑影响/外排功能 使用各种荧光标记的大分子的测定（AIM 3A）。接下来，是否受损 涌入/外排功能是由于BM损伤以及周细胞层粘连蛋白的损失如何影响BM 将探索组成/结构（AIM 3B）。在AIM 4中，周神经元中周细胞层粘连蛋白的功能 将研究损伤/神经变性。在此目的中，是否失去了周细层粘连蛋白会导致 神经元损伤/神经退行性将在生化，结构和功能水平上进行研究。 此外，将确定发生神经元损伤/神经退行性的年龄，并 与神经血管功能障碍发生的那样。这项研究成功完成将 阐明周细层粘连蛋白在神经血管功能和神经元中的基本作用 生存/功能，并确定具有治疗潜力的新型分子靶标在阿尔茨海默氏病中 和阿尔茨海默氏病与疾病有关的痴呆症。此外，该提议也可能导致一代 用于神经变性的创新小鼠模型和新研究的开放门。