Therapy development for open-angle glaucomas

项目摘要 开角型青光眼(OAG)是一组进行性视神经病变，共同是导致 不可逆转的视力丧失。导致其标志-进行性视网膜的致病触发机制 神经节细胞(RGC)死亡，目前尚不清楚，但关键危险因素包括眼压(IOP)和 遗传易感性。眼压升高是由于房水流出阻力的增加。 小梁网状结构(TM)，并与致病细胞外斑块的聚集相关。新疗法 是必要的，因为目前的降眼压治疗并不针对导致眼压下降的病理生理机制 在TM内，进行性视力丧失通常持续存在。我们的长期目标是开发新的、改进的 治疗高压型OAG的机械性治疗策略。这一多学科、协作性 翻译研究将检验这一中心假设，即TM斑块的形成可以被预防或阻止 TM基因表达的选择性修饰。我们将使用腺相关病毒(AAV)作为工具，基于 最近的临床试验成功支持了AAV介导的眼部基因治疗的安全性和有效性。我们的目标是 提供AAV介导的基因替代疗法将预防眼压升高的概念证据 OAG的单基因形式，见于患有Weill-Marchesani综合征(WMS)的ADAMTS10突变人类患者。 具体目标：使用一种成熟的、临床相关的ADAMTS10-OAG犬模型作为测试 平台上，我们提出了三个目标：在目标1中，我们将开发新的衣壳突变AAV载体，以更有效地 在TM中靶向和表达转基因。在目标2中，我们将识别与OAG相关的差异表达基因 在ADAMTS10-突变TM内，并开发它们作为基因治疗的分子生物标记物。在目标3中，我们 将评估AAV后ADAMTS10突变TM细胞基因表达和功能的正常化 介导的ADAMTS10基因替换，并评估它是否提供了长期预防增加的 小梁流出阻力和眼压。意义：基于广泛的初步数据，并使用大型- 以动物OAG模型和ADAMTS10转基因为检测平台，我们将提供具体的证据 新型衣壳突变AAV可以有效地靶向TM内的致病机制，使其正常化 基因表达和生物功能，并提供疾病表型的长期临床救援，包括 小梁流出阻力、眼压升高和RGC丢失。开发这种以机械为基础的疗法将 促进未来的动物研究，为临床试验铺平道路，并增进对 TM内的分子OAG疾病机制。创新：我们的新治疗策略将有选择性地 修饰TM基因表达。我们将创建专门针对TM细胞的新的AAV衣壳载体，以及 通过将临床结果指标与TM转录组的变化相关联来评估治疗效果 独一无二的翻译改进。我们的创新包括我们新颖、成熟的跨学科协作 在成功实施眼部基因治疗的转化研究方面有着良好的记录。

Project Summary Open-angle glaucoma (OAG) is a group of progressive optic neuropathies that together are leading causes of irreversible vision loss. The pathogenic triggering mechanisms that lead to its hallmark, progressive retinal ganglion cell (RGC) death, are unknown, but key risk factors include increased intraocular pressure (IOP) and genetic predisposition. IOP elevation results from increased aqueous humor (AH) outflow resistance via the trabecular meshwork (TM), and correlates with accumulation of pathogenic extracellular plaques. New therapies are needed as current IOP-lowering treatments do not target the contributing pathophysiological mechanisms within the TM, and progressive vision loss often persists. Our long-term goal is to develop novel, improved mechanistic therapeutic strategies to treat high-pressure forms of OAG. This multidisciplinary, collaborative translational study will test the central hypothesis that TM plaque formation can be prevented or halted by selective modification of TM gene expression. We will use adeno-associated virus (AAV) as a tool, based on recent successes in clinical trials supporting safety and efficacy of AAV-mediated ocular gene therapy. We aim to provide proof-of-concept that AAV-mediated gene replacement therapy will prevent IOP elevation in a monogenic form of OAG, seen in ADAMTS10-mutant human patients with Weill-Marchesani syndrome (WMS). Specific Aims: Using a well-established, clinically-relevant canine model of ADAMTS10-OAG as a testing platform, we propose 3 Aims: In Aim 1, we will develop new capsid mutated AAV vectors to more efficiently target and express transgene in the TM. In Aim 2, we will identify OAG-relevant differentially expressed genes within the ADAMTS10-mutant TM and develop them as molecular biomarkers for gene therapy. In Aim 3, we will evaluate the normalization of gene expression and function of ADAMTS10-mutant TM cells following AAV- mediated ADAMTS10-gene replacement, and assess whether it provides long-term prevention of increased trabecular outflow resistance and IOP. Significance: Based on extensive preliminary data and using a large- animal OAG model and the ADAMTS10 transgene as testing platforms, we will provide proof that specific pathogenic mechanisms within the TM can be targeted efficiently with novel capsid mutated AAV, normalizing gene expression and biologic function and providing long-term clinical rescue of disease phenotype, including trabecular outflow resistance, elevated IOP, and RGC loss. Developing such a mechanistic-based therapy will facilitate future animal studies and pave the way towards clinical trials, and also advance understanding of molecular OAG disease mechanisms within the TM. Innovation: Our new therapeutic strategy will selectively modify TM gene expression. We will create new AAV capsid vectors tailored specifically to target TM cells, and assess therapeutic effects by correlating clinical outcome measures with changes in the TM transcriptome, a unique translational advance. Our innovation includes our novel, well-established cross-disciplinary collaborative team with a strong track record in successfully executing translational research in ocular gene therapy.