Ultrasound-assisted extracellular vesicle engineering and induced release: EVEiR

项目总结 而基于纳米颗粒的平台已经成为治疗疾病的领先药物传递平台 各种疾病，目前的纳米医学仍然存在一些挑战，包括毒性、低效 跨越内皮屏障，快速清除，并在体内非特异性蓄积。额外的- 细胞囊泡(EVS)提供了一种天然的递送系统，可以转移各种细胞货物 到相邻和遥远的细胞。电动汽车提供独特的工程优势，同时拥有- 遗传的免疫逃避能力和组织穿透特性。然而，效率低下 治疗性货物包装和细胞产生的电动汽车不足限制了目前基于电动汽车的 药物输送方法。 慢性炎症性疾病(CID)给社区带来了健康和经济负担- 世界各地的联系。目前对CID的治疗既不充分，也不能改变疾病。我们的终极 目标是为CID患者开发一种安全有效的EV疗法。总体目标 本申请中目的是(I)开发一种基于超声(US)的平台，称为EVEiR(Extrael- 叶状囊泡工程和诱导释放)用于传递抗炎细胞因子IL10， 以及(Ii)使用体外肠道模型确定它们的治疗效果。核心假设是-- Esis是使用US刺激和机械本身的外部应用的机械提示- 细胞微环境的联系可能会增加工程化电动汽车的产量和功能 (EEVS)来源于3D培养的间充质干细胞(MSCs)。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 抗炎携带IL10的EEVS(IL10+EEVS)；2)论证IL10+的可行性 从3D培养的骨髓间充质干细胞衍生的EEVS用于靶向治疗。目标一号将决定 使用美国的新技术生产IL10+EEV的效率。目标2将会-- IL-10+EEVS的体外肠道特性及其抗炎作用 模特。这一方案的创新之处在于利用非病毒内毒素的影响发挥协同作用。 EVS中的细胞IL-10包装，脉冲超声刺激MSCs的独特概念 DroGel构建了高效的EEV生产。此外，体外肠道模型允许 生理相关环境中抗炎IL10+EEVS的有效表征- 门槛。拟议的研究具有重要意义，因为该项目的成功完成将 开发快速、经济高效且可扩展的平台，以生成MSC衍生的治疗性电动汽车 用于治疗CID，促进骨髓间充质干细胞来源的电动汽车移植到临床。

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.