Towards personalized medicine in ophthalmology: a new therapeutic approach combining PARP-inhibition and AAV-mediated gene replacement improves vision in advanced stages of retinal neurodegeneration

迈向眼科个性化医疗：结合 PARP 抑制和 AAV 介导的基因替换的新治疗方法可改善视网膜神经退行性晚期阶段的视力

• 批准号: 389782910
• 负责人: Dr. Regine Mühlfriedel
• 金额: --
• 依托单位: Forschungsinstitut für Augenheilkunde
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/389782910?language=en
• 关键词: Towards; personalized; medicine; ophthalmology; new

The combined therapeutic approach outlines a new strategy for a personalized treatment and requires deep insight and knowledge about the major properties and key players leading and guiding the pathological process that is unique for a mutation. This is of particular relevance for Retinitis Pigmentosa (RP), a genetically heterogeneous group of retinal hereditary diseases causing photoreceptor cell death, thus resulting in progressive visual field constriction and finally complete blindness. Despite the advancing genetic knowledge, to date, no effective cure is available for this disease in human patients. Although in genetic diseases, specific restoration of gene function would seem like an ideal therapeutic target, in clinical ophthalmologic practice, the genetic heterogeneity and often late diagnosis of RP could be a drawback for this particular approach. Thus, there is a need for the development of mutation-independent therapeutic strategies that are applicable regardless of the primary genetic defect. In this context, our proposal provides a new combination of therapeutic approaches for RP animal models by using a mutation-independent treatment with PARP-inhibitors or AAV-siRNA-PARP (neuroprotection) to (1) open the therapeutic window for AAV-based gene delivery (gene replacement) and to (2) restore the function of the mutated protein at more advanced stages of degeneration. Based on our findings from Objective 1, we will study the therapeutic efficacy of a new innovative approach combining PARP inhibition followed by an AAV-mediated gene delivery (Objective 5). In Objective 2, we plan to figure out the endpoint for a curative AAV-mediated treatment defined by the causative mutation properties and related molecular mechanisms. In Objectives 3 and 4, the most effective concentration of a PARP inhibitor (drugs or AAV-siRNA-PARP vector) will be evaluated indicating the optimal point of halting photoreceptor cell death. In Objective 5, the most exciting and promising part, we aim at addressing to different issues combining results based on Objectives 1-4. The five objectives will be translated into the practical work by five work packages; the neuroprotection including in vitro and in vivo treatments as well all project related immunhistological, biochemical investigations and assays will be performed by Dr. Sahaboglu Tekgöz. The treatment by rAAV-mediated gene and investigations in vivo by using non-invasive diagnostic techniques on different animal models for RP will be performed by Dr. Mühlfriedel. In terms of the outcome, comparison and interaction of experimental data, all objectives are closely related and give reason to expect two benefits: a) delay in photoreceptor degeneration including cone photoreceptor survival for lifetime and b) restoration of protein function and morphology at more advanced stages when the point of -no return- is exceeded.

联合治疗方法概述了个性化治疗的新策略，需要对主要特性和领导和指导突变独特病理过程的关键参与者有深刻的见解和知识。这与视网膜色素变性（RP）特别相关，RP是一组遗传异质的视网膜遗传性疾病，引起感光细胞死亡，从而导致进行性视野收缩并最终完全失明。尽管遗传知识不断进步，但迄今为止，人类患者中这种疾病没有有效的治疗方法。虽然在遗传性疾病中，基因功能的特异性恢复似乎是一个理想的治疗靶点，但在临床眼科实践中，RP的遗传异质性和通常的晚期诊断可能是这种特定方法的缺点。因此，需要开发不依赖于突变的治疗策略，无论原发性遗传缺陷如何都适用。在这种情况下，我们的提议通过使用PARP抑制剂或AAV-siRNA-PARP（神经保护）的突变非依赖性治疗为RP动物模型提供了新的治疗方法组合，以（1）打开基于AAV的基因递送（基因置换）的治疗窗口，以及（2）在更晚期的变性阶段恢复突变蛋白质的功能。基于我们在目标1中的发现，我们将研究一种新的创新方法的治疗效果，该方法将PARP抑制与AAV介导的基因递送相结合（目标5）。在目标2中，我们计划通过致病突变特性和相关分子机制确定治愈性AAV介导治疗的终点。在目标3和4中，将评估PARP抑制剂（药物或AAV-siRNA-PARP载体）的最有效浓度，指示停止感光细胞死亡的最佳点。在目标5中，最令人兴奋和有希望的部分，我们旨在解决不同的问题，结合目标1-4的结果。这五个目标将通过五个工作包转化为实际工作;神经保护包括体外和体内治疗以及所有项目相关的免疫组织学，生化研究和测定将由Sahaboglu Tekgöz博士进行。Mühlfriedel博士将通过rAAV介导的基因进行治疗，并使用非侵入性诊断技术在不同的RP动物模型上进行体内研究。在实验数据的结果、比较和相互作用方面，所有目标都密切相关，并有理由预期两个益处：a）延迟光感受器变性，包括视锥光感受器终身存活，和B）当超过不可逆点时，在更晚期阶段恢复蛋白质功能和形态。