Suppression of Myocilin by siRNA

siRNA 对 Myocilin 的抑制

• 批准号: 6852544
• 负责人: Andrew J.W. Huang
• 金额: $ 14.95万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6852544
• 关键词: RNA interference; cell death; cell proliferation; cytotoxicity; gene delivery system; gene induction /repression; gene therapy; glaucoma; myocilin; nanotechnology; particle; plasmids; small interfering RNA; tissue /cell culture; trabecular meshwork; transfection /expression vector

DESCRIPTION (provided by applicant): Glaucoma is 1 of the leading causes of blindness in the world. It affects approximately 66.8 million people worldwide. At least 12,000 Americans are blinded by this disease each year. Recent studies have linked mutations of myocilin, a 55Kd secretory glycoprotein to the pathogenesis of open-angle glaucoma in certain patients. It is believed that accumulation of misfolded mutant myocilins induces cell death in trabecular meshwork (TM), which causes obstruction of TM with increased resistance to aqueous outflow and leads to elevated intraocular pressure with eventual optic nerve damage in open-angle glaucoma. We hypothesize that suppressing the accumulation of mutant myocilin proteins in TM cells, thus preventing cytotoxic effects and cell death, could mitigate myocilin-related open-angle glaucoma. In this proposed research, we will use RNA interference via our custom-designed small interfering RNAs (siRNA; 21-23 nucleotides) to modulate, or suppress, the expression of wild-type and mutant myocilin proteins in cultured 293 and TM cells. The siRNA-mitigated TM cytotoxicity (induced by mutant myocilin) will be studied. We will also explore the feasibility of using a novel nanoencapsulation technology to optimize the delivery of siRNAs into the TM cells for specific suppression of myocilin. Taking advantage of the high specificity of siRNAmediated myocilin suppression and the efficient transfection of nanoencapsulated particles via the ocular surface, this proposal will invesitigate the combination of these 2 recent scientific breakthroughs, i.e., siRNA and nanoencapsulation, as a potential glaucoma therapy. Results from these experiments will provide the foundation for future animal studies and could help in the development of potential therapeutic modalities for myocilin-related glaucoma or other inherited ocular disorders.

描述（由申请人提供）：青光眼是世界上主要的致盲原因之一。它影响到全世界约6680万人。每年至少有12，000名美国人因这种疾病而失明。最近的研究表明，在某些开角型青光眼患者中，55 Kd分泌性糖蛋白myocilin的突变与开角型青光眼的发病机制有关。据信，错误折叠的突变型肌球蛋白的积累诱导小梁网（TM）中的细胞死亡，这导致TM阻塞，增加了对房水流出的阻力，并导致眼内压升高，最终导致开角型青光眼中的视神经损伤。我们假设，抑制突变型肌球蛋白在TM细胞中的积累，从而防止细胞毒性作用和细胞死亡，可以减轻肌球蛋白相关的开角型青光眼。在这项拟议的研究中，我们将通过我们定制设计的小干扰RNA（siRNA; 21-23个核苷酸）来调节或抑制野生型和突变型myocilin蛋白在培养的293和TM细胞中的表达。将研究siRNA减轻的TM细胞毒性（由突变型肌球蛋白诱导）。我们还将探索使用新的纳米封装技术来优化siRNA向TM细胞中的递送以特异性抑制肌球蛋白的可行性。利用siRNA介导的肌球蛋白抑制的高特异性和通过眼表面的纳米封装颗粒的有效转染，该提议将研究这两个最近的科学突破的组合，即，siRNA和纳米囊封，作为一种潜在的青光眼治疗。这些实验的结果将为未来的动物研究提供基础，并有助于开发肌球蛋白相关青光眼或其他遗传性眼部疾病的潜在治疗方法。