Nanofabricated Devices and Nanomedicine Approaches for Wound Healing

项目摘要/摘要 这项有指导的研究提案旨在建立一个工程师和纳米科学家的实验室 生物医学研究。这项提议的科学焦点是组织纳米转基因(TNT)，一种电动 将质粒、RNA和寡核苷酸运送到活体组织的基因转移技术。这 技术是第一个实现皮肤到功能血管的非病毒体内再编程的技术(IV， 诱导血管发生)或电生理活性神经细胞(IN，诱导神经发生)。 使用纳米电穿孔方法进行免疫存活，这种方法不会造成基因组整合或 转型。TNT有助于将重新编程的货物包装成称为exosome的生物纳米载体。 虽然这种方法的可行性已经在小鼠皮肤上进行了测试，但它在临床前环境中的应用是 还有待检验。建议的工作集中在TNT芯片的新设计上，该芯片可以用于发布 体内内源性工程外切体，用于糖尿病小鼠和人类的治疗目的。是这样的 采用的方法确保了较高的临床意义。该项目包括以下三个目标。？瞄准 1：纳米电穿孔诱导细胞释放外切体。？1.1：纳米电穿孔增加细胞 Rab-GTP酶在微管上的细胞骨架重组和募集 身体向富含肌动蛋白的皮质释放外切体。？1.2：纳米电穿孔可以作为一种方法 包装内源性信使核糖核酸和miRNA，通过外切体释出。？目标2：测试组织的功效 纳米转基因诱导的工程化外切体加速糖尿病创面闭合。？2.1：质粒的设计 用于体内细胞特异性的信使核糖核酸包装。？2.2：通过TNT将体内细胞特异性mRNA包装在外切体内 加速糖尿病伤口愈合。目的3：研制人体规模的TNT2.1芯片，用于初步的临床前试验。

Project Summary/Abstract This mentored research proposal seeks to aims establish a laboratory of an engineer and nanoscientist in biomedical research. The scientific focus of this proposal is on tissue nanotransfection (TNT), an electromotive gene transfer technology that delivers plasmids, RNA and oligonucleotides to the live tissue in vivo. This technology was the first to achieve non-viral in vivo reprograming of the skin to functional blood vessels (iV, induced vasculogenesis) or to electrophysiologically active neural cells (iN, induced neuritogenesis) under immune survillence using a nanoelectroporation approach that does not pose the risk of genomic integration or transformation. TNT facilitates packaging of reprogramming cargo into biological nano carrier called exosome. Although the feasibility of this approach has been tested in murine skin, its application in pre-clinical setting is yet to be tested. The proposed work focuses on a new design of TNT chip that can be leveraged for release of endogenous engineered exosomes in vivo for therapeutic purposes in both diabetic mice and humans. Such approach was adopted to ensure high clinical significance. The project includes the following three aims. ?Aim 1: Nanoelectroporation induces release of exosome from cells. ?1.1: Nanoelectroporation increases cellular cytoskeletal reorganization and recruitment of Rab-GTPase at the microtubules for transfer of multivesicular bodies to the actin-rich cortex for release of exosome. ?1.2: Nanoelectroporation can be used as a method to package endogenous mRNA and miRNA for release via exosome. ?Aim 2: Test the efficacy of tissue nanotransfection induced engineered exosome in accelerating diabetic wound closure. ?2.1: Design of plasmids for cell-specific packaging of mRNA in vivo. ?2.2: In vivo cell specific mRNA packaging within exosome via TNT accelerates diabetic wound healing. ?Aim 3: Develop human scale TNT2.1 chip for preliminary preclinical testing.