Gene Gun Technology, Opioids, and Corneal Diseases

描述：(申请者摘要)角膜上皮调节液体 运输正常的基质水化和角膜透明度，并作为 防止眼部感染的解剖学和生理学屏障。这 上皮处于不断更新的状态，而对该上皮的损伤 需要及时重新浮出水面，以重建视觉功能。工具用于 在分子水平上实现对角膜疾病的研究和临床应用 水平提供了令人兴奋的晋升途径，但由于需要 操纵基因活动。通过基因治疗，基因治疗取得了进展 一种颗粒介导的基因转移传递系统(基因枪)的研制 这项技术被证明是一种快速、高效和无毒的方法 体内和体外的基因转染法。在这笔赠款中，我们建议 建立有价值的角膜上皮研究工具--基因枪 生物学，获得基因枪作为一种 用于临床治疗的设备，并记录交互的重要性 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动态平衡的系统 人和动物眼表上皮的愈合。具体的 目标包括：1.确定OGF-OGFr接口的重要性 大鼠角膜上皮细胞的维持 OGF受体基因和蛋白活性的分子扰动。2.定义 内源性阿片系统在大鼠角膜创伤愈合中的作用 表皮生长因子受体基因和蛋白的遗传改变 在大鼠体内的表达。3.明确OGF和OGFr功能的意义 人角膜上皮器官培养在伤口愈合过程中的作用 转基因组织产生过量或缺乏OGF受体的组织。 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输送系统。此外，这些 调查将同时获得关于 内源性阿片系统与角膜相关的分子基础 角膜上皮自稳与术后角膜完整性的恢复 受伤。最终，这样的数据可以用来设计分子策略来 矫正视力障碍。

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.