Gene Gun Technology, Opioids, and Corneal Diseases

描述：（申请人摘要）角膜上皮调节液体 正常基质水合作用和角膜透明度的运输，并充当 对抗眼部感染的解剖学和生理学屏障。这个 上皮处于不断更新的状态，并且上皮受到损伤 需要立即重新表面以重新建立视觉功能。工具 在分子水平上实现角膜疾病的研究和临床应用 级别提供了令人兴奋的晋升途径，但由于需要 操纵基因活性。基因治疗已取得进展 开发粒子介导的基因转移传递系统（基因枪） 该技术被证明是一种快速、高效、无毒的方法 体内和体外基因转染。在这笔赠款中，我们建议 建立基因枪作为角膜上皮有价值的研究工具 生物学，获得基因枪作为一种药物的功效的初步信息 临床治疗设备，并记录相互作用的重要性 天然抑制肽阿片生长因子 (OGF) 及其受体 （OGFr）在分子水平上作为调节体内平衡的系统 人类和动物眼表上皮的愈合。具体的 目标包括： 1. 确定 OGF-OGFr 连接的重要性 通过检查后果来维持大鼠角膜上皮 OGF 受体基因和蛋白质活性的分子扰动。 2. 定义 内源性阿片类系统在大鼠角膜伤口愈合中的作用 OGF 受体基因和蛋白质遗传改变后的上皮细胞 在大鼠中的表达。 3. 确定OGF和OGFr功能的意义 在人角膜上皮伤口愈合过程中使用器官培养 组织经过基因改造，产生过量或缺乏的 OGF 受体。 从这些研究中获得的信息将有助于取得突破 了解角膜表面的基因传递系统。而且，这些 调查将同时获得有关该问题的宝贵知识 内源性阿片类药物系统与角膜相关的分子基础 上皮稳态以及术后角膜完整性的恢复 受伤。最终，这些数据可以用来设计分子策略 治疗视力障碍。

DESCRIPTION: (Applicant's Abstract) The corneal epithelium modulates fluid transport for normal stromal hydration and corneal transparency, and serves as an anatomical and physiological barrier against ocular infection. This epithelium is in a constant state of renewal, and injury to this epithelium requires prompt resurfacing in order to re-establish visual function. Tools to achieve research and clinical applications to corneal diseases at the molecular level offer exciting avenues for advancement, but are stymied by the need for manipulating gene activity. Progress has been made in gene therapy through the development of a particle-mediated gene transfer delivery system (gene gun), a technique shown to be a rapid, highly efficient, and nontoxic method for transfection of genes both in vivo and in vitro. In this grant we propose to establish the gene gun as a valuable research tool in corneal epithelial biology, obtain preliminary information for the efficacy of the gene gun as a device for clinical treatment, and document the importance of the interaction of a native inhibitory peptide, opioid growth factor (OGF) with its receptor (OGFr) at the molecular level as a system regulating the homeostasis and healing of the ocular surface epithelium in humans and animals. The specific aims include: 1. Determine the importance of OGF-OGFr interfacing on maintenance of rat corneal epithelium by examining the consequences of molecular perturbation of OGF receptor gene and protein activity. 2. Define the role of an endogenous opioid system related to wound healing of the rat corneal epithelium following genetic alteration of the OGF receptor gene and protein expression in rat. 3. Ascertain the significance of OGF and OGFr function during wound healing of the human corneal epithelium using organ culture of tissues genetically modified to yield an excess or deficit of OGF receptor. Information derived from these studies will contribute to a breakthrough understanding of gene delivery systems to the corneal surface. Moreover, these investigations will simultaneously acquire invaluable knowledge about the molecular basis by which an endogenous opioid system relates to corneal epithelial homeostasis, and the restoration of corneal integrity following injury. Ultimately, such data can be employed to design molecular strategies to remedy visual dysfunction.