权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Human Astrocyte-Based Nanovesicles to Target Neuroinflammation in Alzheimer's Disease

基于人星形胶质细胞的纳米囊泡可针对阿尔茨海默病的神经炎症

基本信息

批准号：
10348978
负责人：
Robert Conrad Krencik
金额：
$ 44.41万
依托单位：
METHODIST HOSPITAL RESEARCH INSTITUTE
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：
2022-02-15 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10348978
关键词：
3-Dimensional Adhesions Adrenal Cortex Hormones Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Amyloid beta-Protein Anti-Inflammatory Agents Antibodies Area Astrocytes Back Binding Biological Assay Biomedical Engineering Biomimetics Calcium Cell Adhesion Molecules Cell Separation Cell physiology Cells Charge Clinical Coculture Techniques Data Development Dexamethasone Drug Targeting Early identification Endothelium Flow Cytometry Fluorescence Formulation Foundations Future Gene Expression Profiling Generations Genetic Goals Homeostasis Human Image Image Cytometry Imaging Device Impaired cognition Inflammation Inflammatory Intervention Knowledge Laboratories Lead Leukocytes Lipids Measures Mediating Membrane Membrane Proteins Microglia Microscopy Nanotechnology Nerve Degeneration Nervous system structure Neuroglia Neuronal Dysfunction Neurons Neurophysiology - biologic function Neurosciences Outcome Pathway interactions Pharmaceutical Preparations Property Proteins Proteomics Protocols documentation Research Signal Transduction Signaling Molecule Source Surface System Technology Testing Therapeutic Therapeutic Agents Time Translating abeta oligomer aged base cell community cell type effective therapy exosome human pluripotent stem cell improved inhibitor innovation innovative technologies insight monolayer nanovesicle neural network neuroinflammation neuropathology next generation novel optical imaging relating to nervous system specific biomarkers stability testing targeted delivery targeted imaging targeted treatment theranostics tool uptake

项目摘要

PROJECT SUMMARY-ABSTRACT Astrocytes are specialized glial cells that are highly abundant in the nervous system and that maintain functional homeostasis of neural networks. In the aged and the Alzheimer’s disease brain, astrocytes can contribute inflammatory signaling molecules to the surrounding micro-environment, in turn, negatively impacting neural function. How can therapeutics, such as anti-inflammatories, be selectively delivered to these dysfunctional astrocytes in order to reduce off-target drug effects on other cells? At present, no effective clinical approaches exist for this purpose. Here, we aim to overcome this lack of technology by formulating lipid nanovesicles capable of enhanced targeted delivery. Development of this innovative technology will be enabled through the combined expertise of two synergistic laboratories who will bioengineer membrane proteins from human pluripotent stem cell-derived astrocytes into the surface of lipid-based nanovesicles. Our preliminary studies have revealed that nanovesicles integrated with membrane proteins derived from unique cell sources retain unique cell adhesion proteins that may lead to cell-specific targeting. This finding provoked our hypothesis that nanovesicles coated with adhesion molecules derived from Alzheimer’s disease-model astrocytes (a.k.a., AstroVesicles (AVs)) will bind to protein-interacting partners, specifically at the surface of inflammatory astrocytes and, thus, increase cellular uptake by dysfunctional astrocytes. In this way, AVs could be a potential new theranostic tool that allows for the early identification of inflamed areas as well as the delivery of therapeutic cargo. Astrocyte inflammation will be induced by amyloid beta oligomer treatment to model the Alzheimer’s disease microenvironment and then reactivity will be confirmed by functional calcium imaging and gene expression profiling. To test the hypothesis, in Aim 1, we will formulate nanovesicles and compare the size, charge, and stability of those containing membrane proteins from naïve and oligomer-treated inflammatory astrocytes as well as from other sources (e.g., neurons, microglia, and cell-derived exosomes). We will perform proteomics-based discovery of the nanovesicles to identify cell-specific proteins with high potential for cell targeting based on known cell-cell protein interactions. In Aim 2, we will validate the expected capability of AVs to preferentially target astrocytes that are inflamed via amyloid beta oligomer treatment, in comparison to naïve astrocytes and microglia. We will also test potential mechanisms of targeting by interfering with candidate proteins identified in preliminary data. Our approach will be to measure the presence of AVs upon treatment of sphere cultures composed of astrocytes, neurons, and microglia, using three-dimensional optical imaging and flow cytometry. Notably, these studies will pioneer the use of human neural spheres for nanovesicle testing. Finally, in Aim 3, we will test whether AVs yield enhanced functional delivery of anti-inflammatories, focusing on the NFB pathway. After optimizing and validating the AVs, we expect this innovative system will be utilized throughout the neuroscience community for cell-targeted therapeutics and imaging tools in Alzheimer’s disease.

项目摘要星形胶质细胞是神经系统中高度丰富的特化胶质细胞，神经网络的功能稳态。在老年人和阿尔茨海默病患者的大脑中，星形胶质细胞可以促进炎症信号分子周围的微环境，反过来，神经功能如何选择性地将治疗药物，如抗炎药，功能失调的星形胶质细胞，以减少脱靶药物对其他细胞的影响？目前尚无有效的临床存在用于此目的的方法。在这里，我们的目标是克服这种缺乏技术，能够增强靶向递送的纳米囊泡。这项创新技术的发展将使通过两个协同实验室的综合专业知识，他们将生物工程膜蛋白，将人多能干细胞衍生的星形胶质细胞植入基于脂质的纳米囊泡的表面。我们的初步研究表明，与来自独特细胞来源的膜蛋白整合的纳米囊泡保留独特的细胞粘附蛋白，可能导致细胞特异性靶向。这一发现激发了我们的假设用源自阿尔茨海默病模型星形胶质细胞（a.k.a.， AstroVesicles（AV））将与蛋白质相互作用的伴侣结合，特别是在炎症表面。星形胶质细胞，并因此增加功能失调的星形胶质细胞的细胞摄取。这样一来，自动驾驶汽车可能成为一种潜在的一种新的治疗诊断工具，可以早期识别发炎区域并提供治疗药物，货物.星形胶质细胞炎症将通过淀粉样蛋白β寡聚体治疗诱导以模拟阿尔茨海默氏症疾病微环境，然后反应性将通过功能性钙成像和基因表达谱分析为了检验这一假设，在目标1中，我们将配制纳米囊泡并比较其大小，电荷和稳定性的那些含有膜蛋白从幼稚和寡聚体治疗的炎症星形胶质细胞以及其它来源（例如，神经元、小胶质细胞和细胞衍生的外泌体）。我们将执行基于蛋白质组学的纳米囊泡发现，以确定具有高细胞增殖潜力的细胞特异性蛋白质。基于已知的细胞-细胞蛋白质相互作用的靶向。在目标2中，我们将验证无人驾驶汽车的预期能力与未经治疗的星形胶质细胞相比，星形胶质细胞和小胶质细胞。我们还将通过干扰候选人来测试潜在的靶向机制。在初步数据中发现的蛋白质。我们的方法将是测量AV治疗后的存在，使用三维光学成像技术，流式细胞仪值得注意的是，这些研究将开创使用人类神经球进行纳米囊泡测试的先河。最后，在目标3中，我们将测试AV是否产生增强的抗炎药功能性递送，重点是 NF κ B B通路。经过优化和验证的自动驾驶汽车，我们希望这个创新的系统将被利用在整个神经科学界的细胞靶向治疗和成像工具在阿尔茨海默病。