A Nanocarrier Platform for Targeting Schlemm's Canal Cells

项目摘要 纳米级药物载体（即纳米载体）由于其转运多种药物的能力而引起了广泛的关注。 治疗剂和诊断剂以及选择性靶向特定细胞和组织。这增加了特异性 可以有显着的临床意义，包括减少副作用和更高的剂量， 力量Schlemm氏管（SC）内皮细胞有望成为青光眼治疗的细胞靶点，因为它们在治疗青光眼中的作用。 机械刚度与眼内压（IOP）的调节有关。Rho激酶抑制剂和肌动蛋白- 解聚剂降低内皮细胞硬度，并显著降低动物和人的IOP， 其中几种已被批准用于临床。然而，这些药剂与显著的副作用相关， 包括结膜充血和角膜轮动。研究表明，>50%的患者接受这些治疗， 治疗剂显示出不利的副作用。靶向纳米载体递送系统可以解决这些问题， 目前不能通过角膜上皮，因此必须通过 眼内注射由于频繁的眼内注射不会被患者很好地耐受，因此持续的眼内注射是不可能的。 需要递送系统来最小化药物施用的频率。基因治疗靶点 低眼压的治疗已经出现，为未来的青光眼治疗带来了希望， 眼内注射，但需要靶向基因递送系统来增强SC细胞的选择性转染。 因此，存在对持续纳米载体递送系统和基因递送系统两者的显著需求 针对SC的眼内策略。考虑到这些需求，本提案的目标是设计 一个可扩展的，可定制的，合成的纳米载体平台，可以适用于运输不同的治疗药物， 药物以可控的释放速率释放到流出途径细胞。这项工作的成功完成将导致 第一个用于持续眼内释放纳米载体的递送系统，一种新型的非病毒基因递送平台 选择性转染SC细胞，并完成非人灵长类动物研究，以证明这些临床试验的合理性。 人体内的输送系统。 将完成以下具体目标： 目的1：优化靶向施累姆氏管细胞的纳米载体的治疗效果的持续时间，同时避免 副作用和毒性在小鼠眼睛的角膜和血管组织内。 目的2：证明使用靶向纳米载体在体内非病毒转染施累姆氏管细胞，而不需要使用靶向纳米载体。 影响小鼠的邻近眼组织 目的3：证明含有拉曲库林-A的靶向纳米载体显著增加了常规的细胞毒性。 流出设施和降低非人灵长类动物的IOP，无不良反应。

PROJECT SUMMARY Nanoscale drug carriers (i.e. nanocarriers) have attracted much attention for their ability to transport diverse therapeutic and diagnostic agents and to selectively target specific cells and tissues. This increased specificity can have significant clinical implications, including decreased side effects and lower dosages with higher potency. Schlemm's canal (SC) endothelial cells hold promise as a cellular target for glaucoma therapy, as their mechanical stiffness is associated with modulation of intraocular pressure (IOP). Rho kinase inhibitors and actin- depolymerizing agents reduce endothelial cell stiffness and significantly lower IOP in animals and humans with several now approved for clinical use. However, these agents are associated with significant side effects, including conjunctival hyperemia and corneal verticillata. Studies show that >50% of patients treated with these therapeutics exhibit adverse side effects. Targeted nanocarrier delivery systems may address these issues but are not currently capable of passing through the corneal epithelium and must therefore be administered via intraocular injection. As frequent eye injections would not be well tolerated by patients, sustained intraocular delivery systems are needed to minimize the frequency of drug administration. Gene therapy targets for treatment of ocular hypotension have emerged, holding promise for a future glaucoma cure following a single intraocular injection, but a targeted gene delivery system is needed to enhance selective transfection of SC cells. A significant need therefore exists for both sustained nanocarrier delivery systems and gene delivery systems for intraocular strategies targeting the SC. With these needs in mind, the objective of this proposal is to engineer a scalable, customizable, synthetic nanocarrier platform that can be adapted to transport diverse therapeutic agents to outflow pathway cells with controllable release rates. Successful completion of this work will result in the first delivery system for sustained intraocular release of nanocarriers, a novel nonviral gene delivery platform for selective transfection of SC cells, and completion of nonhuman primate studies to justify clinical trials of these delivery systems in humans. The following Specific Aims will be completed: Aim 1: Optimize the duration of therapeutic effect for nanocarriers targeting Schlemm’s canal cells while avoiding side effects and toxicity within the cornea and vascular tissues in mouse eyes. Aim 2: Demonstrate nonviral transfection of Schlemm's canal cells in vivo using targeted nanocarriers without affecting nearby ocular tissues in mice Aim 3: Demonstrate that targeted nanocarriers containing latrunculin-A significantly increase conventional outflow facility and lower IOP in nonhuman primates without adverse effects.