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，它们一起是青光眼的主要原因。 不可逆转的视力丧失致病触发机制导致其标志，进行性视网膜 神经节细胞（RGC）死亡，是未知的，但关键的风险因素包括眼内压（IOP）升高， 遗传倾向IOP升高是由于通过眼内动脉的房水（AH）流出阻力增加所致。 小梁网（TM），并与致病性细胞外斑块的积累。新疗法 由于目前的降眼压治疗并不针对促成的病理生理机制， 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的机械治疗策略。这种多学科的，合作的 一项转化研究将检验中心假设，即TM斑块的形成可以通过以下方法预防或停止： 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 最近在临床试验中的成功支持了AAV介导的眼部基因治疗的安全性和有效性。我们的目标 为了提供AAV介导的基因替代疗法将预防眼内压升高的概念验证， OAG的单基因形式，见于患有Weill-Marchesani综合征（WMS）的ADAMTS 10突变型人类患者。 特定目的：使用已建立的临床相关ADAMTS 10-OAG犬模型作为试验 平台，我们提出了3个目标：在目标1中，我们将开发新的衣壳突变的AAV载体，以更有效地 在TM中靶向并表达转基因。在目标2中，我们将鉴定OAG相关的差异表达基因 在ADAMTS 10突变体TM中，并将它们开发为基因治疗的分子生物标志物。在目标3中， 将评估在AAV-100转染后ADAMTS 10-突变TM细胞的基因表达和功能的正常化。 介导的ADAMTS 10基因替代，并评估其是否提供了长期预防增加的 小梁流出阻力和IOP。重要性：基于广泛的初步数据，并使用大量的- 动物OAG模型和ADAMTS 10转基因作为测试平台，我们将提供证据， TM内的致病机制可以用新的衣壳突变的AAV有效地靶向， 基因表达和生物学功能，并提供疾病表型的长期临床挽救，包括 小梁流出阻力、IOP升高和RGC损失。开发这种基于机制的疗法将 促进未来的动物研究，为临床试验铺平道路，并促进对 TM内的分子OAG疾病机制。创新：我们的新治疗策略将选择性地 改变TM基因表达。我们将创建专门针对TM细胞的新AAV衣壳载体， 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， 独特的翻译进展。我们的创新包括我们新颖的，完善的跨学科合作 在成功执行眼部基因治疗的转化研究方面有着良好记录的团队。

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.