Surgical Microneedle Patch Delivery of CMMP for Heart Repair

项目摘要 基于细胞的治疗代表了再生医学中一种有前途的策略。然而，活细胞需要 在使用前仔细保存和处理。此外，作为“活药”，细胞移植携带一定的 免疫原性和/或致瘤性风险。无细胞和无生命疗法的发展具有 有可能彻底改变目前的再生医学实践。越来越多的证据表明， 干细胞主要通过分泌促再生因子发挥其有益作用。在此基础上， PI Cheng实验室通过封装干细胞分泌的 生物可降解聚合物块中的因子。我们之前的研究表明，这些“合成细胞” 携带着与亲代细胞相似的分泌蛋白和细胞膜。在小鼠心肌梗死模型中， 心肌梗死（MI）时，心肌内注射CMMP可保护存活心肌， 类似于细胞疗法的心脏功能。尽管概念验证成功，但一个巨大的挑战是 将这些治疗性微粒有效地递送到心脏。心脏补片已被测试， 将治疗细胞移植到心脏表面。一个警告是，由于缺乏补丁-主机通信， 心脏补片与宿主心肌的整合不良。MPI Gu实验室在以下方面经验丰富： 微针（MN）贴片的制造。我们以前的研究表明，MN贴片可以提供治疗 有效地对组织。目前的R 01提案代表了我们以前工作的逻辑进展， 带来新技术。在这里，我们将开发和测试一种新的实体：MN-CMMP心脏贴片 通过将CMMP嵌入可生物降解和生物相容的微针心脏贴片形成。此外，本发明还提供了一种方法， 我们的研究将从以前的啮齿动物急性心肌梗死模型扩展到小/大的慢性心肌梗死模型， 动物总体假设是MN-CMMP可以进一步改善CMMP治疗的疗效， 慢性心脏损伤的大鼠和猪。目的1：制备由微针贴片组成的MN-CMMP 测定MN-CMMP在培养细胞中的体外效力。目标2：确定安全性和 MN-CMMP治疗在慢性MI大鼠模型中功效。目标3：将发现转化为临床相关 猪心脏损伤模型。我们的研究将为创新和“现成”疗法奠定基础 基于可从供体干细胞系标准化的分泌因子和心肌基质， 异种心脏组织我们的方法的无细胞性质更容易转化为临床。 虽然这个特殊的拨款申请的目标是心脏和心脏干细胞，我们的方法代表了一个 平台技术，可用于创建多种类型的合成干细胞和器官 用于修复各种其他器官的基质。

PROJECT SUMMARY Cell-based therapy represents a promising strategy in regenerative medicine. However, live cells need to be carefully preserved and processed before usage. In addition, as “live drugs”, cell transplantation carries certain immunogenicity and/or tumorigenicity risks. The development of cell-free and non-living therapeutics has the potential to revolutionize current regenerative medicine practice. Mounting lines of evidences indicate that adult stem cells exert their beneficial effects mainly through the secretion of pro-regenerative factors. Based on this, the PI Cheng Lab fabricated cell mimicking microparticles (CMMPs) by encapsulating stem cell-secreted factors in a biodegradable polymer block. Our previous studies demonstrated that those “synthetic cells” carried similar secreted proteins and membranes as their parental cells did. In a mouse model of myocardial infarction (MI), intramyocardial injection of CMMPs led to preservation of viable myocardium and augmentation of cardiac functions similar to cell therapy. Despite the successful proof of concept, a big challenge is the effective delivery of those therapeutic microparticles to the heart. Cardiac patches have been tested to deliver therapeutic cells to the surface of the heart. One caveat is that there is a lack of patch-host communication due to poor integration of the cardiac patch with the host myocardium. The MPI Gu Lab is experienced in the fabrication of microneedle (MN) patches. Our previous studies indicated that MN patch can deliver therapeutics to the tissue effectively. The present R01 proposal represents a logic progression from our previous work while bringing new technologies. Here we will be developing and testing a new entity: a MN-CMMP cardiac patch formed by embedding CMMPs into biodegradable and biocompatible microneedle cardiac patches. In addition, our studies will extend from the previous rodent acute MI model to a chronic MI model in both small/large animals. The overarching hypothesis is that MN-CMMP can further improve the efficacy of CMMP therapy in rats and pigs with chronic heart injury. AIM 1: Fabricate MN-CMMP comprised of microneedle patch loaded with CMMPs; Determine in vitro potency of MN-CMMP in cultured cells. AIM 2: Determine the safety and efficacy of MN-CMMP therapy in rat model of chronic MI. AIM 3: Translate the findings into clinically-relevant porcine models of heart injury. Our study will form the foundation for an innovative and “off the shelf” therapy based on secreted factors and myocardium matrix that can be standardized from donor stem cell lines and xenogenic cardiac tissues. The cell-free nature of our approach is more readily translatable to the clinic. Although this particular grant application targets the heart and cardiac stem cells, our approach represents a platform technology that can be applied to the creation of multiple types of synthetic stem cell and organ matrices for the repair of various other organs.