Improving the throughput of diagnosis and treatment of inherited diseases of the retina

项目摘要/摘要 尽管遗传性视网膜变性(IRD)的基因检测取得了进展，但检测到 未知重要性(VUS)可阻止患者接受基因诊断。中国的长期目标是 建议的研究是使用基于细胞的分析来解决这个问题，这种分析可以有效地识别哪些DNA 变异是致病的突变，是良性的多态，其规模将导致 医学上可采取行动的信息。红斑狼疮是导致视力丧失的重要原因，越来越多的人可以通过 基因特异性治疗，如基因增强治疗。虽然准确的基因诊断是至关重要的 在实施基因特异性治疗之前，确定特定患者的遗传原因 IRD可能很困难，大约三分之一的患者完全无法接受基因诊断。因此，在那里 是前所未有的需要有效地识别IRD的遗传因果关系，以便将现有的和 新出现的保护视力或恢复视力的疗法。为了满足这一需要，拟议的 研究是通过一系列综合目标来利用这一机会，重点是有效地识别 重要IRD基因的致病变异。拟议的研究试图改变当前的研究 范式--分析IRD基因中发现的少量DNA变异--到范式 在那里，关于医学上重要的IRD基因的变异的大量数据被提前生成。 因此，这项拟议的研究验证了经验性的、基于细胞的分析可以有效地用于 并准确识别人类中哪些DNA变体是致病的并导致IRDS，哪些可能是 良性多态。在目标1中，我们汇集并描述了潜在的 重要的显性IRD基因视紫红质致病氨基酸的变化。这一目标的扩展测试 这些突变中的哪些可以接受小分子伴侣疗法。在《目标2》中，这些技术 都经过修改，以表征一个潜在的致病氨基酸变化的全面集合 重要的隐性IRD基因，RPE65。进一步假设，将化验结果与人类进行比较 表型数据将定义与人类致病结果相对应的适当数值范围。观影 总而言之，这些目标提供了一条途径，以产生一个开放可用的资源，可以立即提供 向医学遗传学家、遗传咨询师和研究人员提供有关IRDS中VU的高保真信息 红斑狼疮。它利用了一个重要的机会，在那里对目标的调查可以直接产生 医学上可采取行动的信息，从而将治疗提供给原本无法治疗的患者。

Project Summary/Abstract Despite advances in genetic testing for inherited retinal degenerations (IRDs), detection of a DNA variant of unknown significance (VUS) can prevent a patient from receiving a genetic diagnosis. The long-term goal of the proposed research is to address this problem using cell-based assays that can efficiently identify which DNA variants are disease-causing mutations and which are benign polymorphisms, at a scale that would produce medically-actionable information. IRDs are important causes of vision loss, and are increasingly treatable by gene-specific therapies such as gene augmentation therapy. While an accurate genetic diagnosis is critical before administering a gene-specific therapy, confident identification of the genetic cause for particular patient’s IRD can be difficult, with about one third of patients failing to receive a genetic diagnosis altogether. Thus, there is an unprecedented need to efficiently identify the genetic causality of IRDs in order to translate existing and emerging sight-preserving or sight-restoring therapies to patients. To address this need, the goal of the proposed research is to capitalize on this opportunity via a set of integrated Aims focused on the efficient identification of pathogenic variants in important IRD genes. The proposed research seeks to shift the current research paradigm-- analyzing small numbers of DNA variants in IRD genes as they are discovered-- to a paradigm where large quantities of data are generated in advance about variants in medically-important IRD genes. Therefore, the proposed research tests the hypothesis that empiric, cell-based assays can be used to efficiently and accurately identify which DNA variants in humans are pathogenic and cause IRDs, and which are likely benign polymorphisms. In Aim 1, we assemble and characterize a comprehensive collection of potentially pathogenic amino acid changes in an important dominant IRD gene, rhodopsin. An expansion of this Aim tests which of these mutations are amenable to chaperone therapy with small molecules. In Aim 2, these techniques are modified to characterize a comprehensive collection of potentially pathogenic amino acid changes in an important recessive IRD gene, RPE65. It is further hypothesized that comparing assay results to human phenotype data will define proper numerical ranges which correspond to pathogenic results in humans. Viewed together, these Aims provide a pathway for producing an openly-available resource that could instantly provide higher-fidelity information about VUS in IRDs to medical geneticist, genetic counselors, and investigators of IRDs. It takes advantage of a significant opportunity where investigation of the Aims can directly produce medically-actionable information resulting in the delivery of therapies to otherwise untreatable patients.