Gene Gun Technology, Opioids, and Corneal Diseases

基因枪技术、阿片类药物和角膜疾病

• 批准号: 6641233
• 负责人: IAN S ZAGON
• 金额: $ 14.05万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6641233
• 关键词: biotechnology; cornea disorder; corneal epithelium; endogenous opioid; evaluation /testing; gene delivery system; gene therapy; genetic transcription; histochemistry /cytochemistry; homeostasis; human tissue; laboratory rat; nonhuman therapy evaluation; opioid receptor; organ culture; technology /technique development; western blottings; wound healing; biotechnology; cornea disorder; corneal epithelium; endogenous opioid; evaluation /testing; gene delivery system; gene therapy; genetic transcription; histochemistry /cytochemistry; homeostasis; human tissue; laboratory rat; nonhuman therapy evaluation; opioid receptor; organ culture; technology /technique development; western blottings; wound healing

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.

描述：（申请人的摘要）角膜上皮调节流体 运输正常的基质水合和角膜透明度，并用作 针对眼部感染的解剖学和生理障碍。这 上皮处于恒定状态，并且对此上皮的损伤 需要及时重新铺面才能重新建立视觉功能。工具 在分子的角膜疾病中实现研究和临床应用 级别提供了令人兴奋的进步途径，但受到需要的需求。 操纵基因活性。通过基因治疗取得了进展 开发粒子介导的基因转移递送系统（Gene Gun），A 证明是一种快速，高效且无毒的方法 在体内和体外转染基因。在这笔赠款中，我们建议 建立基因枪作为角膜上皮的宝贵研究工具 生物学，获得基因枪功效的初步信息 用于临床治疗的设备，并记录相互作用的重要性 及其受体的天然抑制性肽，阿片类药物生长因子（OGF） （OGFR）在分子水平上，作为调节体内平衡和的系统 人类和动物中眼表上皮的愈合。具体 目的包括：1。确定OGF-OGFR接口的重要性 通过检查大鼠角膜上皮的维护 OGF受体基因和蛋白活性的分子扰动。 2。定义 与大鼠角膜伤口愈合有关的内源性阿片类药物系统的作用 OGF受体基因和蛋白质的遗传改变后上皮 大鼠的表达。 3。确定OGF和OGFR功能的重要性 在人类角膜上皮的伤口愈合期间，使用器官培养 基因修饰的组织以产生过量或OGF受体的不足。 这些研究得出的信息将有助于突破 了解基因输送系统到角膜表面。而且，这些 调查将同时获得有关该研究的宝贵知识 内源性阿片类药物与角膜相关的分子基础 上皮稳态，以及角膜完整性的恢复 受伤。最终，可以将这些数据用于设计分子策略 补救视觉功能障碍。