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

项目摘要 拟议的研究项目解决了生物工程方法的需要， 可促进心肌梗死后组织修复的基于靶向外泌体的治疗递送平台 （密歇根州）。在临床前和临床环境中积累的证据一致表明， 从成体干细胞/基质细胞分泌的细胞因子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。基于外泌体的疗法代表了从常规疗法的范式转变， 以细胞为基础的心血管治疗方法。然而，目前的基于外泌体的治疗剂遭受以下问题： 存在几个问题，包括缺乏靶向性、装载额外治疗剂的复杂制造工艺 货物和费用此外，使得能够控制递送基于外泌体的产物的技术已经被广泛应用。 尚待探索。为了克服这些挑战，我们建议开发一种智能心脏起搏器， 纳米胶囊（nanocaptantine，CXo）用于靶向和控制递送间充质干细胞衍生的 外泌体（MSC-Exo）和治疗性蛋白质以促进MI后的心脏修复。我们的主要假设是， (1)用外来体和心脏保护蛋白（例如，前动力蛋白2，聚集蛋白， 胰岛素样生长因子1，骨髓衍生生长因子）可以通过静电逐层工程化 与新型酶介导的自由基聚合（EMRP）技术相结合的组装过程;以及（2） 所述的ESTXo能够响应于基质的上调而递送组合治疗剂 金属蛋白酶（MMPs）是心肌缺血损伤的标志，并触发修复信号通路 通过治疗性蛋白质介导修复缺血性心脏。拟议中的XuanXo战略的创新 包括（1）靶向损伤的心肌细胞;（2）以非创伤性方式递送组合治疗剂的能力， 需求时尚，通过利用缺血心脏中的心脏病理学;（3）双重再生机制， 靶向心脏修复，结合了类似于干细胞治疗的外泌体介导的旁分泌机制 心脏保护蛋白介导的信号通路。我们将制造并优化 负载有各种心脏保护蛋白，并评估BACTEXo的体外生物活性。安全性， 将在患有以下疾病的大鼠中研究CXO治疗的功能益处和潜在作用模式： 缺血性心脏损伤成功地执行拟议的研究项目将建立一个温和和轻便的 外泌体功能化的方法，并形成创新和现成的外泌体的基础- 基于智能和有针对性的纳米医学的治疗，用于MI后心脏的修复和再生。 虽然这个特殊的拨款申请的目标是心脏，但XiamXo战略代表了一个平台， 该技术可推广到多种组织的修复和再生。

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.