Clickable Extracellular Vesicles to Silk-Based Biomaterials for Regenerative Medicine

项目摘要 成功的再生医学方法需要利用适当的细胞信号， 正确的时间来指导宿主组织的功能。这些信号通常是由自然再生的 在发育和稳态过程中由间充质干细胞（MSC）控制的过程， 他们分泌的细胞外囊泡（EV），这允许一种基于细胞但无细胞的方法， 下游再生技术。这个多学科的MPI提案汇集了一个团队， 两名资深研究人员领导再生医学重点小组，优势互补， 共同研究者与关键角色，以及工业合作伙伴RoosterBio，Inc.我们将共同创造， 测试一种创新的使能技术，以稳定地将EV结合到用于组织的生物材料中， 工程和再生医学应用。具体来说，我们将使用一种新型的叠氮化物基点击 化学技术可控地将EVs转化为丝素蛋白作为示范应用， 也可以在其它生物材料、基质或表面，甚至组织上进行固定。我们 鉴于其FDA批准的状态和广泛的用途，我们选择丝作为本申请中的生物材料。 我们假设“叠氮化物可点击”MSC衍生的EV（我们将简称为“Az-EV”） 将比未改性的EV更稳定地固定到丝素蛋白生物材料上，这将导致 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 -证明和验证Az-EV固定到丝纤蛋白基材料; 目的2 -证明体外固定到丝上的Az-EV的MSC模拟效果; 目的3 -证明Az-EV在小鼠慢性伤口愈合模型中的再生作用; 目的4 -证明Az-EV在大鼠组织工程血管移植模型中的再生作用。 合作伙伴RoosterBio，Inc.将MSC衍生的Az-EV的生产“工业化”（扩大规模）， 商业化，以提供给其他研究人员和临床医生。 这项研究将为这种新型的选择性EV固定化的有效性提供见解 在感兴趣的生物系统内有效地引导EV递送的技术。我们的概念验证 研究将证明如何利用这种再生技术可以帮助治疗慢性 并使TEVG具有改善的通畅率。

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.