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

这种联合治疗方法概述了个性化治疗的新战略，需要对主要属性和关键参与者的深刻洞察力和知识，以领导和指导突变所特有的病理过程。这与视网膜色素变性(RP)尤其相关，RP是一组遗传异质性的视网膜遗传性疾病，导致光感受器细胞死亡，从而导致进行性视野收缩，最终导致完全失明。尽管有先进的遗传知识，但到目前为止，还没有有效的治疗方法来治疗人类患者的这种疾病。虽然在遗传性疾病中，特定的基因功能恢复似乎是一个理想的治疗目标，但在临床眼科实践中，遗传异质性和通常延迟诊断可能是这种特殊方法的缺点。因此，有必要开发不依赖于突变的治疗策略，这些策略适用于任何主要的遗传缺陷。在此背景下，我们的建议为RP动物模型提供了一种新的治疗方法组合，通过使用PARP抑制剂或AAV-siRNA-PARP(神经保护)的非突变治疗来(1)打开基于AAV的基因传递(基因替换)的治疗窗口，以及(2)在更高级的退变阶段恢复突变蛋白的功能。基于我们在目标1中的发现，我们将研究一种结合PARP抑制和AAV介导的基因传递的新的创新方法的治疗效果(目标5)。在目标2中，我们计划找出由致病突变特性和相关分子机制定义的AAV介导的治疗性治疗的终点。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，以指示阻止光感受器细胞死亡的最佳时间点。在目标5，这是最令人兴奋和最有希望的部分，我们的目标是结合目标1-4的成果，处理不同的问题。这五个目标将通过五个工作包转化为实际工作；包括体外和体内治疗在内的神经保护以及所有与项目相关的免疫学、生化调查和分析将由Sahaboglu Tekgöz博士执行。通过rAAV介导的基因治疗和使用非侵入性诊断技术对不同动物模型进行体内研究将由MühlFriedel博士执行。就实验数据的结果、比较和相互作用而言，所有目标都是密切相关的，并有理由预期有两个好处：a)延缓光感受器退化，包括视锥感光细胞终身存活；b)当超过不返回点时，在更高级的阶段恢复蛋白质的功能和形态。

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.