Exosome-Based Pathological Microenvironment-Responsive Nanocapsules for Targeted Heart Repair

项目总结 拟议的研究项目解决了对生物工程方法的需求，以实现智能和 可促进心肌梗死后组织修复的靶向外切体治疗给药平台 (Mi)。在临床前和临床环境中积累的证据一致地表明，外切体 由成体干细胞/基质细胞分泌，协调细胞治疗的主要作用模式，以修复 心肌梗死后心肌损伤。基于外显子的疗法代表着从传统的 基于细胞的心血管治疗方法。然而，目前基于外切体的治疗方法受到 几个问题，包括缺乏靶向性，复杂的制造工艺来加载额外的治疗 货物和费用。此外，能够控制外周体基产品递送的技术已经 还有待探索。为了克服这些挑战，我们建议开发一种智能心脏疗法 用于靶向和控制输送间充质干细胞来源的纳米胶囊(MedicoXo) 外切体(MSC-Exo)和促进心肌梗死后心脏修复的治疗蛋白。我们的中心假设是 (1)带有外切体和心脏保护蛋白(例如，原激动素2，集聚蛋白， 胰岛素样生长因子1，髓系衍生生长因子)可以通过逐层静电工程来实现 装配工艺与一种新的酶介导自由基聚合(EMRP)技术相结合；以及(2) 心脏Xo可使联合治疗药物的交付响应于基质的上调 金属蛋白酶(MMPs)，心肌缺血损伤的标志，并触发修复信号通路 由治疗性蛋白介导修复缺血心脏。心动Xo战略的创新 包括(1)靶向受损的心肌细胞；(2)在On-1中提供联合治疗的能力- 在缺血心脏中利用心脏病理改变的需求方式；(3)双再生机制 靶向心脏修复结合外体介导的旁分泌机制，类似于干细胞治疗 与心脏保护蛋白介导的信号通路有关。我们将制造和优化MedicoXo 负载各种心脏保护蛋白，并评估心脏Xo的体外生物活性。安全， 心脏Xo疗法的功能益处和潜在的作用模式将被研究在 缺血性心脏损伤。拟议研究项目的成功实施将建立一个温和而便捷的 探讨外切体功能化并形成创新和现成的外切体的基础-- 基于智能和靶向纳米药物的心肌梗死后心脏修复和再生的基础治疗。 尽管这一特殊的赠款申请针对的是心脏，但MedicoXo战略代表着一个平台 可推广到多种组织修复和再生的技术。

PROJECT SUMMARY The proposed research project addresses the need for a bioengineering approach to a smart and targeted exosome-based therapeutic delivery platform that can promote tissue repair after myocardial infarction (MI). Accumulating evidence in both preclinical and clinical settings has consistently indicated the exosomes secreted from adult stem/stromal cells orchestrate the principal modes of action of cell therapy to repair the injured heart muscle following MI. Exosome-based therapeutics represent a paradigm shift from conventional cell-based approaches to cardiovascular therapy. However, current exosome-based therapeutics suffer from several problems including lack of targetability, complicated fabrication process to load additional therapeutic cargos, and expense. In addition, technologies that enable controlled delivery of exosome-based products have yet to be explored. To overcome these challenges, we propose to develop a smart cardiotherapeutic nanocapsule (CardioXo) for the targeted and controlled delivery of both mesenchymal stem cell-derived exosomes (MSC-Exo) and therapeutic proteins to promote heart repair post-MI. Our central hypotheses are that (1) the CardioXo functionalized with both exosomes and cardioprotective proteins (e.g., prokineticin 2, agrin, insulin-like growth factor 1, myeloid-derived growth factor) can be engineered via an electrostatic layer-by-layer assembly process combined with a novel enzyme-mediated radical polymerization (EMRP) technology; and (2) the CardioXo can enable the delivery of combination therapeutics in response to the upregulation of matrix metalloproteinases (MMPs), a hallmark of cardiac ischemic injury, and trigger the reparative signaling pathways mediated by therapeutic proteins to repair the ischemic heart. The innovation of the proposed CardioXo strategy includes (1) targeting to the injured cardiomyocytes; (2) ability to deliver combination therapeutics in an on- demand fashion by leveraging cardiac pathology in the ischemic heart; (3) dual-regenerative mechanism for targeted heart repair that combines the exosome-mediated paracrine mechanism similar to stem cell therapy with the cardioprotective protein-mediated signaling pathways. We will fabricate and optimize the CardioXo loaded with various cardioprotective proteins and assess the in vitro bioactivity of CardioXo. The safety, functional benefits, and the potential modes of action of CardioXo therapy will be investigated in rats with ischemic heart injury. Successful execution of the proposed research project will establish a mild and facile approach to exosome functionalization and form the foundation of an innovative and off-the-shelf exosome- based therapy based on smart and targeted nanomedicine for the repair and regeneration of the post-MI heart. Although this particular grant application targets the heart, the CardioXo strategy represents a platform technology that can be generalized to the repair and regeneration of multiple tissues.