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.

项目概要/摘要 该应用的长期目标是了解基底膜（BM）、 神经血管单元的非细胞成分，参与阿尔茨海默病的发病机制 疾病，并可以有针对性地治疗阿尔茨海默病和阿尔茨海默病相关的痴呆症。 这与NIA的使命是一致的，该提案旨在研究其生物学功能。 周细胞层粘连蛋白在：（1）神经血管功能，包括血脑屏障（BBB）完整性、脑 血流 (CBF) 和脑流入/流出功能，以及 (2) 神经元存活/功能 在目标 1 中， 首先，将调查周细胞层粘连蛋白在 BBB 完整性中的功能是否丧失以及程度如何。 周细胞层粘连蛋白对 BBB 完整性的影响将使用各种分子的 FITC-Dextrans 进行研究 接下来，周细胞层粘连蛋白诱导 BBB 损失的分子机制。 将研究分解，重点是细胞旁和跨细胞运输的变化 此外，介导周细胞层粘连蛋白作用的受体。 将鉴定和检查内皮细胞（目标 1C）。在目标 2 中，周细胞层粘连蛋白的功能。 首先，将使用以下方法研究周细胞层粘连蛋白的损失如何影响 CBF。 接下来，定量放射自显影和双光子成像（目标 2A）是否减少了 CBF。 将在体外和体内研究由周细胞损失/变性引起的（目标 2B）。 将鉴定并检查介导周细胞层粘连蛋白作用的受体（目标 2C）。 目标 3，将研究周细胞层粘连蛋白在脑流入/流出功能中的作用，首先，是否和。 将通过流入/流出来研究周细胞层粘连蛋白的损失如何影响大脑流入/流出功能 使用各种荧光标记的大分子进行分析（目标 3A），以确定是否受损。 流入/流出功能是由于 BM 损伤以及周细胞层粘连蛋白的损失如何影响 BM 将探讨神经元中周细胞层粘连蛋白的组成/结构（目标 3B）。 在此目的中，将研究周细胞层粘连蛋白的损失是否会导致损伤/神经变性。 神经元损伤/神经变性将在生化、结构和功能水平上进行研究。 此外，将确定神经损伤/神经变性发生的年龄并 与神经血管功能障碍发生时相比，本研究的成功完成将。 阐明周细胞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，并确定具有阿尔茨海默病治疗潜力的新分子靶点 此外，这一提议还可能导致与阿尔茨海默病相关的痴呆症的产生。 神经退行性变的创新小鼠模型的研究，为新研究打开了大门。