Suppression of Myocilin by siRNA

siRNA 对 Myocilin 的抑制

• 批准号: 7051986
• 负责人: Andrew J.W. Huang
• 金额: $ 14.6万
• 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/7051986
• 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 one 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 two 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(申请人提供)：青光眼是世界上导致失明的主要原因之一。它影响着全球约6680万人。每年至少有12000名美国人因这种疾病而失明。最近的研究表明，肌球蛋白(一种55Kd的分泌性糖蛋白)的突变与某些患者开角型青光眼的发病机制有关。据认为，在开角型青光眼中，错误折叠的突变型myocilins的积聚导致小梁网(trabecular network，TM)细胞死亡，从而导致TM受阻，房水流出阻力增加，导致眼压升高，最终导致视神经损伤。我们推测，抑制突变的Myoclin蛋白在TM细胞中的积累，从而防止细胞毒效应和细胞死亡，可以减轻Myoclin相关的开角型青光眼。在这项拟议的研究中，我们将使用RNA干扰通过我们定制的小干扰RNA(siRNA；21-23个核苷酸)来调节或抑制培养的293和TM细胞中野生型和突变型myoclin蛋白的表达。将研究siRNA减轻TM细胞毒性(由突变的myoclin诱导)。我们还将探索使用一种新的纳米封装技术来优化将siRNAs输送到TM细胞中以特异性抑制Myoclin的可行性。利用siRNA介导的抑制肌球蛋白的高度特异性和通过眼表有效地导入纳米包裹颗粒的优势，这一建议将探索这两项最新科学突破的组合，即siRNA和纳米胶囊，作为潜在的青光眼治疗方法。这些实验的结果将为未来的动物研究提供基础，并可能有助于开发与霉菌素相关的青光眼或其他遗传性眼病的潜在治疗方式。

项目成果

Suppression of keratoepithelin and myocilin by small interfering RNAs (siRNA) in vitro.

• 发表时间: 2007-11
• 期刊: Molecular vision
• 影响因子: 2.2
• 作者: Ching Yuan;E. J. Zins;A. Clark;A. Huang

Suppression of keratoepithelin and myocilin by small interfering RNA (an American Ophthalmological Society thesis).

• 发表时间: 2007
• 期刊: Transactions of the American Ophthalmological Society
• 作者: A. Huang