Optimizing Rho-associated kinase inhibitors for use in treating Fuchs endothelial corneal dystrophy

项目摘要/摘要 Fuchs内皮性角膜营养不良(FECD)是一种多基因疾病，影响超过610万美国人 40岁，是美国角膜移植手术的主要适应症。FECD的诊断基于 角膜内皮细胞可见损伤及细胞外基质变性的研究 角膜(滴眼)。FECD的发病机制以CEC功能障碍、内皮细胞到间充质细胞 过渡(EMT)、细胞外基质(ECM)退化和CEC死亡导致视力丧失。尽管它 可以早期诊断，FECD需要角膜移植治疗视力丧失，因为没有治疗方法 阻止它的发展。我们先前发现，Rho相关激酶抑制物(RKI)瑞帕舒地尔减少 FECD表型，值得探索其作为预防疾病的可能治疗选择的潜力 移植。目前，两种RKI(奈塔舒地尔和瑞帕舒地尔)正被用作辅助治疗以加速 无内皮细胞角膜移植(DWEK)的后裂术中内皮伤口的愈合。研究还没有 已经进行了这两个RKI的头对头比较。RKI治疗的问题包括眼睛刺激 以及频繁给药的要求。此外，RKI还没有作为FECD的预防性药物进行测试 进步。拟议研究的总体目标是确定两个RKI的相对有效性 (netarsudil和rigasudil)，用于治疗FECD(Dwek，主要药物)的两种不同用途 预防FECD进展的治疗)。我们的中心假设是奈扎舒地尔和雷帕舒地尔的疗效相当 伤口愈合和FECD表型逆转的有效性以及将RKI包装成靶向纳米粒 (NPS)将减少非目标副作用，并允许减少剂量频率。建议的理由是 研究包括以下内容：i)需要进行正面的RKI比较，因为如果netarsudil有效，那么 临床使用可能更快；2)需要减少剂量和缓释策略 因为眼睛刺激限制了RKI的使用；以及iii)需要测试RKI抑制FECD表型 确定是否有必要进行未来的临床试验。在强劲的初步数据的指导下，我们的假设将得到检验 通过以下具体目标：1)确定FECD DWEK治疗模型中RKIs的最佳剂量； 2)确定在FECD中使用和不使用靶向NP包装的RKI的临床疗效和毒性 疾病预防小鼠模型。该方法的创新之处在于它使用了一种新的细胞培养模型，使 有可能描述与早发性FECD疾病进展相关的病理事件以及 新型纳米颗粒，能够以更高的效率和更低的毒性将药物输送到FECD细胞。这个 拟议的研究意义重大，因为RKI的使用将允许不依赖于 供体角膜组织或角膜移植(全球每70个所需供体角膜中只有1个供体角膜可用)。 这项建议的风险更高，因为RKI是否可以防止FECD疾病进展尚不清楚，但有 开发可能推迟或阻止手术的药物治疗方法的高潜在回报。

PROJECT SUMMARY/ABSTRACT Fuchs endothelial corneal dystrophy (FECD) is a polygenic disease that affects 6.1 million Americans over 40 years of age and is the leading indication for corneal transplant surgery in the U.S. FECD is diagnosed based on visible damage to corneal endothelial cells (CECs) and degenerative extracellular matrix deposits on the inner cornea (guttae). FECD pathogenesis is characterized by CEC dysfunction, endothelial-to-mesenchymal transition (EMT), extracellular matrix (ECM) degeneration, and CEC death that lead to vision loss. Although it can be diagnosed early, FECD requires corneal transplantation for vision loss because there are no therapies to prevent its progression. We found previously that the Rho-associated kinase inhibitor (RKI) ripasudil decreases FECD phenotypes, warranting an exploration of its potential as a possible therapeutic option to prevent transplantation. Currently, two RKIs (netarsudil and ripasudil) are being used as adjuvant therapies to accelerate endothelial wound healing in Descemetorhexis without endothelial keratoplasty (DWEK). Studies have not yet been performed comparing these two RKIs head-to-head. Problems with RKI treatments include ocular irritation and frequent dosing requirements. Additionally, RKIs have not been tested as a preventative for FECD progression. The overall objectives of the proposed research are to determine the relative efficacy of two RKIs (netarsudil and ripasudil) for use in two distinct applications for the treatment of FECD (DWEK, primary drug therapy to prevent FECD progression). Our central hypothesis is that netarsudil and ripasudil will have equivalent efficacy in wound healing and FECD phenotype reversal, and that packaging RKIs into targeted nanoparticles (NPs) will decrease off-target side effects and permit reduced dosing frequency. The rationale for the proposed research includes the following: i) head-to-head RKI comparisons are needed because if netarsudil works then clinical use may be achieved more quickly; ii) dose-reduction and sustained-release strategies are needed because ocular irritation limits RKI use; and iii) testing RKIs for suppression of FECD phenotypes is needed to determine if future clinical trials are warranted. Guided by strong preliminary data, our hypothesis will be tested through the following specific aims: 1) Determine the optimum dosing of RKIs in a FECD DWEK treatment model; and 2) Determine the clinical efficacy and toxicity of RKIs with and without targeted NP packaging in a FECD disease prevention mouse model. The approach is innovative in its use of a new cell culture model that makes it possible to delineate the pathological events related to disease progression in early-onset FECD, as well as novel nanoparticles that are capable of delivering drugs to FECD cells with more efficiency and less toxicity. The proposed research is significant because RKI use would permit FECD treatment alternatives that do not rely on donor corneal tissue or corneal transplantation (only 1 donor cornea is available for every 70 needed worldwide). This proposal is of higher risk because it is unknown if RKIs can prevent FECD disease progression, but has a high potential reward of developing drug therapy treatments that may delay or prevent surgery.