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.

项目摘要提取星形胶质细胞是专门的神经胶质细胞，在神经系统中高度丰富并且维持神经网络的功能稳态。在老年人和阿尔茨海默氏病大脑中，星形胶质细胞可以反过来促进周围微环境的炎症信号分子。神经功能。如何选择治疗，例如抗炎药功能失调的星形胶质细胞以减少对其他细胞的脱靶药物影响？目前，没有有效的临床为此目的存在方法。在这里，我们的目标是通过制定脂质来克服这种缺乏技术能够增强目标递送的纳米媒体。将启用这种创新技术的开发通过两个协同实验室的联合专业知识，这些实验室将来自人多能干细胞衍生的星形胶质细胞进入基于脂质的纳米层的表面。我们的初步研究表明，与源自独特细胞源的膜蛋白整合的纳米植物保留可能导致细胞特异性靶向的独特细胞粘附蛋白。这一发现激发了我们的假设那是涂有阿尔茨海默氏病模型星形胶质细胞的粘合分子（又称，也是果蝇（AVS））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，因此增加了功能失调的星形胶质细胞的细胞摄取。这样，AV可能是一种潜力新的Theranotic工具，允许早期鉴定发炎的区域以及提供治疗货物。星形胶质细胞炎症将由淀粉样蛋白β低聚物治疗引起，以建模阿尔茨海默氏症疾病微环境，然后通过功能性钙成像和基因证实反应性表达分析。为了检验假设，在AIM 1中，我们将制定纳米层并比较大小，来自幼稚和低聚物治疗的炎症的膜蛋白的电荷和稳定性星形胶质细胞以及其他来源（例如神经元，小胶质细胞和细胞衍生的外泌体）。我们将表演基于蛋白质组学的发现纳米孢子以鉴定细胞特异性蛋白具有很高的细胞潜力基于已知的细胞蛋白相互作用的靶向。在AIM 2中，我们将验证AV的预期能力优先针对通过淀粉样蛋白β低聚物治疗发炎的星形胶质细胞，与幼稚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靶向的潜在机制初步数据中鉴定出的蛋白质。我们的方法是在处理时测量AV的存在由星形胶质细胞，神经元和小胶质细胞组成的球体培养物，使用三维光学成像和流式细胞仪。值得注意的是，这些研究将开拓人类神经球的使用进行纳米层测试。最后，在AIM 3中，我们将测试AVS是否产生抗炎药的功能传递，重点关注 NFB路径。在优化和验证AV之后，我们希望将使用这种创新系统通过神经科学界的细胞靶向疗法和阿尔茨海默氏病的成像工具。