Engineered extracellular vesicles as a targeted drug delivery system for multiple sclerosis

工程细胞外囊泡作为多发性硬化症的靶向药物递送系统

基本信息

批准号：
10387930
负责人：
Rachel Mizenko
金额：
$ 3.92万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10387930
关键词：
Address Area Autoimmune Biodistribution Biological Assay Biological Availability Biomimetics Blocking Antibodies Blood - brain barrier anatomy Cell Differentiation process Cells Clinical Trials Coculture Techniques Data Demyelinations Development Differentiation Inducer Disease Disease Progression Drug Delivery Systems Drug Targeting Dyes Electroporation Endothelium Engineering Exhibits Experimental Autoimmune Encephalomyelitis Extracellular Protein Fluorescence Goals Heterogeneity Immune Targeting In Vitro Individual Inflammation Knowledge Label Laboratories Lesion Liposomes Mediating Medical Membrane Proteins Methods MicroRNAs Modeling Molecular Multiple Sclerosis Myelin National Research Service Awards Neuraxis Neurodegenerative Disorders Neuroglia Neurologic Neurology Oligodendroglia Pathogenesis Pathway interactions Pattern Pharmaceutical Preparations Pharmacology Population Proteins Recovery Reporting Resources Sorting - Cell Movement Source Surface System Technical Expertise Techniques Testing Therapeutic Tissues Training Universities Untranslated RNA Vesicle blood-brain barrier crossing blood-brain barrier penetration blood-brain barrier permeabilization brain endothelial cell career cell type clinical translation delivery vehicle density detection limit effective therapy extracellular vesicles in vivo in vivo evaluation interest nanoparticle nanoparticle drug next generation novel therapeutics oligodendrocyte precursor oligodendrocyte progenitor precursor cell prevent protein expression receptor remyelination stem cells synthetic drug therapeutically effective trafficking transcytosis translational medicine uptake

项目摘要

Abstract The ultimate goal of this F31 Ruth L. Kirchstein NRSA is to request support to address a fundamental gap in knowledge preventing engineering of efficacious drug delivery vehicles for progressive multiple sclerosis (PMS). PMS is a common, debilitating neurodegenerative disease that causes widespread demyelination in the central nervous system. There are currently no therapies that reliably remediate the advance of PMS, but an emerging strategy is to promote recovery by initiating differentiation of oligodendrocyte progenitor cells (OPCs) to oligodendrocytes, the myelin producing cells depleted in PMS. Current drug delivery techniques to achieve remyelination are either poorly efficacious or highly invasive, major impediments to clinical translation. An effective remyelinating therapeutic for PMS must cross the intact blood-brain barrier (BBB) and then target OPCs. This proposal focuses on the synthesis and testing of engineered extracellular vesicles (eEVs) as a drug delivery vehicle to accomplish these feats. Natural EVs have been shown to both target specific cells/tissues and also cross endothelial barriers. However, due to their immense functional heterogeneity, these qualities do not occur in the same EVs. Although a population of EVs that efficiently targets OPC has been identified, the best EV population and key proteins that promote BBB crossing remain unknown, preventing new biomimetic engineering strategies. To address the limitations of current therapeutics, I propose to identify and fuse subpopulations of EVs that efficiently cross the BBB and target OPCs, load them with microRNA-219, an OPC differentiating agent, to engineer a bioavailable and selective drug delivery vehicle. Building on results obtained in my preliminary data, I will carry out this project in three steps: (1) to optimize a method of fluorescence activated vesicle sorting to identify and isolate BBB-crossing EVs; (2) to elucidate the molecular mechanisms and key proteins during BBB transcytosis using a functional transwell model; and (3) to produce eEVs via fusion of endogenous EVs and loading with remyelinating therapeutics. We will quantify eEV ability to cross the BBB and target OPCs to initiate differentiation and produce myelin in vitro and in vivo. This project is focused on producing a novel therapeutic uniquely suited to PMS, but this pipeline to engineer EVs with multiple targeted functions could be applied to address drug delivery barriers for many medical problems. This project was developed in parallel with a rigorous training plan to enhance my training and technical skills in the areas of neurology, pharmacology, and translational medicine. This plan will enable my transition to independence as I focus on my long-term goal of pursuing an academic career developing neurological therapeutics. Training will exploit the university’s many resources for professional and educational development.

抽象的这个F31 Ruth L. Kirchstein NRSA的最终目标是请求支持以解决基本差距知识预防有效的药物进行工程，用于进行性多发性硬化症（PMS）。 PMS是一种常见的，令人衰弱的神经退行性疾病，会导致中央宽度脱髓鞘神经系统。目前尚无可靠地纠正PM的疗法，但新兴的疗法策略是通过将少突胶质细胞祖细胞（OPC）分化为促进恢复少突胶质细胞，髓磷脂产生的细胞在PMS中耗尽。当前的药物输送技术再髓效率较差，要么是高度侵入性的临床翻译的主要障碍。对PMS进行有效的透明疗法必须越过完整的血脑屏障（BBB），然后靶向 OPC。该提案重点介绍了工程细胞外蔬菜（EEV）作为药物的合成和测试运送工具来完成这些壮举。天然电动汽车已显示出靶向特定细胞/组织的两个以及跨越内皮障碍。但是，由于其极大的功能异质性，这些特质确实如此不发生在同一电动汽车中。尽管已经确定了有效针对OPC的电动汽车人群，但促进BBB杂交的最佳EV种群和关键蛋白仍然未知，以防止新的仿生工程策略。为了解决当前疗法的局限性，我建议识别和融合有效地越过BBB并靶向OPC的电动汽车的亚群，用microRNA-219加载它们区分剂，以设计生物利用和选择性药物输送车辆。在我的初步数据中获得的结果的基础上，我将以三个步骤执行此项目：（1）优化荧光激活的囊泡排序方法，以识别和分离BBB交叉电动汽车；（2）阐明使用功能性跨韦尔模型，在BBB转介症期间的分子机制和关键蛋白质；（3）到通过融合内源性电动汽车并使用透明度治疗产生EEV。我们将量化EEV 能够越过BBB和靶向OPC来启动分化并在体外和体内产生髓磷脂。这项目的重点是生产一种适合PMS的新型热，但该管道向工程师电动汽车具有多个目标功能可用于解决许多医疗问题的药物输送障碍。该项目与严格的培训计划并行开发，以增强我的培训和技术技能神经病学，药理学和转化医学领域。该计划将使我的过渡到独立，我专注于追求发展神经学的长期目标疗法。培训将探索大学用于专业和教育发展的许多资源。