Clickable Extracellular Vesicles to Silk-Based Biomaterials for Regenerative Medicine

项目总结 成功的再生医学方法需要利用适当的细胞信号 指导宿主组织功能的正确时机。这些信号通常是由自然再生细胞发出的 间充质干细胞(MSCs)控制发育和动态平衡的过程 它们分泌的细胞外小泡(EV)，允许基于细胞但无细胞的方法 下游再生技术。这项多学科的MPI提案汇集了一支 两名资深研究人员领导优势互补的再生医学专题小组， 关键角色的联合调查人员，以及行业合作伙伴RoosterBio，Inc.，我们将共同创建和 测试一项创新的使能技术，将电动汽车稳定地整合到用于组织的生物材料中 工程和再生医学应用。具体地说，我们将使用一种新的基于叠氮的点击 可控制地将EVS固定在丝素蛋白上的化学技术是一个示范应用，但 固定化也可以在其他生物材料、基质或表面，甚至组织上进行。我们 考虑到其FDA批准的地位和广泛的用途，在本申请中选择丝绸作为我们的生物材料。 我们假设“叠氮可点击的”MSC衍生的电动汽车(我们将简称为“Az-EVS”) 对丝素生物材料的固定化比未修饰的EVS更稳定，这将导致 具有更高的再生能力。为了验证这一假设并提供概念验证应用程序，我们 将追求四个具体目标： 目的1-验证Az-EV对丝素蛋白材料的固定化； 目的2-体外研究Az-EVS在真丝上的模拟MSC作用； 目的3-证明Az-EVS在小鼠慢性创面愈合模型中的再生作用； 目的4-证明Az-EVS在大鼠组织工程化血管移植模型中的再生作用。 合作伙伴RoosterBio，Inc.将把MSC衍生的Az-Ev的生产“工业化”(放大)，用于 商业化以供其他研究人员和临床医生使用。 这项研究将为这种新型选择性电动汽车固定化的效果提供洞察力。 在感兴趣的生物系统内有效地指导电动汽车交付的技术。我们的概念验证 研究将展示利用这种再生技术如何帮助治疗慢性 创面和使TEVGs具有更高的通畅率。

PROJECT SUMMARY Successful regenerative medicine approaches require harnessing the appropriate cell signals at the right time to direct host tissue functions. These signals are often informed by the natural regenerative processes controlled during development and homeostasis by mesenchymal stem cells (MSCs) and their secreted extracellular vesicles (EVs), which allow a cell-based yet cell-free approach for downstream regenerative technologies. This multidisciplinary, MPI proposal brings together a team of two senior investigators leading regenerative medicine-focused group with complementary strengths, co-investigators with critical roles, and industrial partner RoosterBio, Inc. Together, we will create and test an innovative enabling technology to stably incorporate EVs to a biomaterial intended for tissue engineering and regenerative medicine applications. Specifically, we will use a novel azide-based click chemistry technique to controllably immobilize EVs to silk fibroin as a demonstrative application, but immobilization can also be done on other biomaterials, substrates, or surfaces, or even tissues. We chose silk as our biomaterial in this application given its FDA-approved status and wide use. We hypothesize that “azide-clickable" MSC-derived EVs (which we will refer to simply as “Az-EVs”) will have more stable immobilization to silk fibroin biomaterials than unmodified EVs, and this will result in higher regenerative potency. To test this hypothesis and provide proof-of-concept applications, we will pursue four specific aims: Aim 1 - Demonstrate and validate Az-EV immobilization to silk fibroin-based materials; Aim 2 - Demonstrate the MSC-mimicking effects of Az-EVs immobilized to silk in vitro; Aim 3 - Demonstrate the regenerative effects of Az-EVs in a mouse chronic wound healing model; Aim 4 - Demonstrate the regenerative effects of Az-EVs in a rat tissue engineered vascular graft model. Partner RoosterBio, Inc. will “industrialize” (scale-up) the production of MSC-derived Az-EVs for commercialization to make available to other researchers and clinicians. This research will provide insight to the efficacy of this novel selective EV immobilization technology to efficiently direct EV delivery within a biological system of interest. Our proof-of-concept studies will demonstrate how utilization of this regenerative technology can aid in treating chronic wounds and enabling TEVGs with improved patency rates.