Suppression of Myocilin by siRNA

描述（由申请人提供）：青光眼是世界上致盲的主要原因之一。它影响着全世界大约6680万人。每年至少有12000名美国人因这种疾病失明。最近的研究将心肌蛋白（一种55Kd的分泌糖蛋白）的突变与某些患者开角型青光眼的发病机制联系起来。我们认为，错误折叠的突变型心肌蛋白的积累可诱导小梁网（TM）细胞死亡，从而导致小梁网阻塞，增加对水流出的阻力，导致眼压升高，最终导致开角型青光眼视神经损伤。我们假设，抑制突变心肌蛋白在TM细胞中的积累，从而防止细胞毒性作用和细胞死亡，可以减轻心肌蛋白相关的开角型青光眼。在本研究中，我们将使用RNA干扰，通过我们定制的小干扰RNA （siRNA； 21-23核苷酸）来调节或抑制培养293和TM细胞中野生型和突变型心肌蛋白的表达。将研究sirna减轻的TM细胞毒性（由突变心肌蛋白诱导）。我们还将探索使用一种新的纳米胶囊技术来优化sirna进入TM细胞以特异性抑制心肌蛋白的可行性。利用siRNA介导的心肌蛋白抑制的高特异性和纳米胶囊化颗粒通过眼表的高效转染，本课题将研究将siRNA和纳米胶囊化这两项最新科学突破结合起来作为一种潜在的青光眼治疗方法。这些实验的结果将为未来的动物实验提供基础，并有助于开发与心肌蛋白相关的青光眼或其他遗传性眼部疾病的潜在治疗方法。

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.