Ultrasound-assisted extracellular vesicle engineering and induced release: EVEiR

超声辅助细胞外囊泡工程和诱导释放：EVEiR

• 批准号: 10506164
• 负责人: Masamitsu Kanada
• 金额: $ 24.39万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10506164
• 关键词: 3-Dimensional; Anti-Inflammatory Agents; Antiinflammatory Effect; Biophysical Process; Blood Circulation; Cell membrane; Cells; Characteristics; Chemicals; Clinic; Clinical Research; Clinical Treatment; Communities; DNA; Detection; Disease; Distant; Drug Delivery Systems; Economic Burden; Encapsulated; Endothelium; Engineering; Environment; Extracellular Matrix; Foundations; Future; Generations; Genes; Goals; Hydrogels; Immune Evasion; In Vitro; Interleukin-10; Intestines; Kidney; Liposomes; Liver; Mammalian Cell; Mechanical Stimulation; Mechanics; Mediating; Membrane Proteins; Mesenchymal Stem Cells; Messenger RNA; Methods; Mission; Modeling; Nanotechnology; Patients; Periodicity; Pharmaceutical Preparations; Physiologic pulse; Physiological; Plasmids; Production; Property; Proteins; Public Health; Research; Safety; Solid; Spleen; Stretching; Surface; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Toxic effect; Transfection; Translating; Translations; Transmembrane Transport; Treatment Efficacy; United States National Institutes of Health; Untranslated RNA; base; chronic inflammatory disease; clinical application; cost effective; cytokine; cytotoxicity; delivery vehicle; disability; extracellular vesicles; health economics; in vivo; innovation; mechanical properties; mechanical signal; nanomedicine; nanoparticle; novel; novel strategies; preclinical study; small molecule; sonoporation; synergism; targeted delivery; therapeutically effective; three dimensional cell culture; ultrasound; vector; vesicular release; viscoelasticity

PROJECT SUMMARY While nanoparticle-based platforms have become a leading drug delivery platform for treating various diseases, several challenges remain in current nanomedicine, including toxicity, inefficient endothelial barrier crossing, rapid elimination, and nonspecific accumulation in the body. Extra- cellular vesicles (EVs) provide a natural delivery system that can transfer various cellular cargo to adjacent and distant cells. EVs offer unique advantages for engineering while possessing in- herent immune evasion capability and tissue penetrating characteristics. However, inefficient therapeutic cargo packaging and insufficient EV production from cells limit the current EV-based drug delivery approach. Chronic inflammatory disease (CID) imposes health and economic burdens on communi- ties worldwide. Current therapy of CID is neither sufficient nor disease-modifying. Our ultimate goal is to develop a safe and effective EV-based therapy for CID patients. The overall objectives in this application are to (i) develop an ultrasound (US)-based platform termed EVEiR (Extracel- lular Vesicle Engineering and induced Release) for delivering anti-inflammatory cytokine IL10, and (ii) determine their therapeutic efficacy using an in vitro intestinal model. The central hypoth- esis is that externally applied mechanical cues using US stimulation and the mechanical proper- ties of the cell microenvironment may increase the production and function of engineered EVs (eEVs) derived from mesenchymal stem cells (MSCs) in 3D cultures. We will test the central hypothesis by pursuing two Specific Aims: 1) Develop US-assisted EVEiR for efficient production of anti-inflammatory IL10-carrying eEVs (IL10+ eEVs); and 2) Demonstrate the feasibility of IL10+ eEVs derived from MSCs in 3D cultures for targeted delivery of therapeutics. Aim 1 will determine the efficiency of IL10+ eEV production using the novel US-based techniques. Aim 2 will charac- terize the properties of IL10+ eEVs and their anti-inflammatory effect using an in vitro intestinal model. The innovation of this proposal is to utilize the synergy with the impact of non-viral intra- cellular IL10 packaging in EVs, the unique concept of pulsed US stimulation of MSCs in 3D hy- drogel constructs for efficient eEV production. In addition, an in vitro intestinal model allows for efficient characterization of anti-inflammatory IL10+ eEVs in a physiologically relevant environ- ment. The proposed research is significant because the successful completion of this project will develop a rapid, cost-effective, and scalable platform to generate MSC-derived therapeutic EVs for treating CID and facilitate the translation of MSC-derived EVs to the clinic.

项目摘要 虽然基于纳米颗粒的平台已经成为治疗糖尿病的领先药物递送平台， 尽管目前的纳米医学在治疗各种疾病方面仍存在一些挑战，包括毒性、效率低下、 内皮屏障穿越、快速消除和体内非特异性蓄积。额外- 细胞囊泡（EV）提供了一种天然的递送系统， 到邻近和远距离的细胞。电动汽车为工程设计提供了独特的优势，同时拥有 遗传免疫逃避能力和组织穿透特性。然而，效率低下 治疗性货物包装和来自细胞的EV生产不足限制了当前基于EV的 药物输送方法。 慢性炎症性疾病（CID）给社区带来健康和经济负担， 联系全球。目前CID的治疗既不充分也不能改善疾病。我们的最终 目的是为CID患者开发一种安全有效的EV治疗方法。总体目标 在本申请中是（i）开发一种基于超声（US）的平台，称为EVEiR（Extracel， 泡囊工程和诱导释放）用于递送抗炎细胞因子IL 10， 和（ii）使用体外肠模型测定它们的治疗功效。中央hypoth- 是外部应用的机械线索，使用美国刺激和机械适当- 细胞微环境的联系可能会增加工程EV的生产和功能 在3D培养物中的源自间充质干细胞（MSC）的eEV。我们将测试中央 通过追求两个具体目标来实现假设：1）开发美国辅助的EVEiR，以实现高效生产 携带抗炎性IL 10的eEV（IL 10 + eEV）的活性;和2）证明IL 10 + eEV的可行性。 在3D培养物中衍生自MSC的eEV用于靶向递送治疗剂。目标1将决定 使用基于美国的新技术生产IL 10 + eEV的效率。目标2将charac- 使用体外肠内实验证实IL 10 + eEV的性质及其抗炎作用。 模型该提案的创新之处在于利用非病毒内的影响的协同作用， EV中的细胞IL 10包装，3D hy中MSC的脉冲US刺激的独特概念， 用于高效eEV生产的drogel构建体。此外，体外肠模型允许 在生理相关环境中有效表征抗炎性IL 10 + eEV， 我是说。这项研究的意义重大，因为该项目的成功完成将 开发一个快速、具有成本效益和可扩展的平台，以生成MSC衍生的治疗性EV 用于治疗CID并促进MSC衍生的EV向临床的转化。