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

• 批准号: 10534137
• 负责人: Rachel Mizenko
• 金额: $ 4.01万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 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/10534137
• 关键词: Address; Area; Autoimmune; Biodistribution; Biological Assay; Biological Availability; Biomimetics; Blocking Antibodies; Blood - brain barrier anatomy; Cell Differentiation process; Cells; Central Nervous System; Clinical Trials; Coculture Techniques; Data; Demyelinations; Development; Differentiation Inducer; Disease; Disease Progression; Drug Delivery Systems; Drug Targeting; Dyes; Education; 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; Neurodegenerative Disorders; Neuroglia; Neurologic; Neurology; Oligodendroglia; Pathogenesis; Pathway interactions; Pattern; Pharmaceutical Preparations; Pharmacology; Population; Proteins; Recovery; Reporting; Resources; Sorting; 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; remediation; remyelination; stem cells; synthetic drug; therapeutically effective; trafficking; transcytosis; translational medicine; uptake

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.

摘要 这款F31 Ruth L. Kirchstein NRSA请求支持，以解决 知识阻止工程的有效药物输送车辆进行性多发性硬化症（PMS）。 经前综合征是一种常见的，使人衰弱的神经退行性疾病，其导致中枢神经系统广泛的脱髓鞘。 神经系统目前还没有可靠的治疗方法来补救PMS的进展，但一种新兴的治疗方法正在出现。 一种策略是通过启动少突胶质祖细胞（OPCs）分化为 少突胶质细胞，在PMS中耗尽的髓鞘产生细胞。目前的药物输送技术，以实现 髓鞘再生的效果差或高度侵入性，这是临床转化的主要障碍。 PMS的有效髓鞘再生治疗必须穿过完整的血脑屏障（BBB），然后靶向 OPCs。这项提案的重点是合成和测试工程细胞外囊泡（eEVs）作为药物 运输工具来完成这些壮举。天然EV已显示靶向特定细胞/组织 并且还穿过内皮屏障。然而，由于其巨大的功能异质性，这些特性 不会发生在相同的电动汽车中。尽管已经鉴定了有效靶向OPC的EV群体， 最好的EV群体和促进BBB穿越的关键蛋白仍然未知，阻止了新的仿生 工程战略。为了解决目前治疗的局限性，我建议识别和融合 有效地穿过BBB和靶向OPC的EV亚群，用microRNA-219（OPC 分化剂，以工程化生物可利用的和选择性的药物递送载体。 基于我的初步数据所获得的结果，我将分三步进行这个项目：（1）优化一个 荧光激活囊泡分选方法鉴定和分离BBB交叉EV;（2）阐明 使用功能性transwell模型的BBB转胞吞过程中的分子机制和关键蛋白;和（3） 通过融合内源性EV并加载髓鞘再生治疗剂来产生eEV。我们将量化eEV 能够穿过BBB和靶向OPCs以启动分化并在体外和体内产生髓鞘。这 该项目的重点是生产一种独特的适合PMS的新型治疗方法，但这种工程电动汽车的管道 具有多种靶向功能的药物递送系统可用于解决许多医学问题的药物递送障碍。 这个项目是与严格的培训计划同时开发的，以提高我的培训和技术技能， 神经学、药理学和转化医学领域。这个计划将使我能够过渡到 独立，因为我专注于我的长期目标，追求学术生涯发展神经 治疗学培训将利用大学的许多资源促进专业和教育发展。