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

项目概要/摘要 尽管遗传性视网膜变性（IRD）的基因检测取得了进展，但检测遗传性视网膜变性的DNA变体仍然是一个难题。 未知意义（VUS）可以阻止患者接受基因诊断。的长期目标 一项提议的研究是使用基于细胞的检测来解决这个问题，这种检测可以有效地识别哪些DNA 变异是致病突变，是良性的多态性，其规模会产生 医学上可行的信息IRD是视力丧失的重要原因，并且越来越多地可通过以下方法治疗： 基因特异性治疗，如基因增强治疗。虽然准确的基因诊断至关重要 在给予基因特异性治疗之前，对特定患者的遗传原因进行可靠的鉴定 IRD可能很困难，大约三分之一的患者完全没有接受基因诊断。因此 是一个前所未有的需要，有效地确定遗传因果关系的IRD，以转化现有的， 为患者提供新兴的视力保护或视力恢复疗法。为了满足这一需求，拟议的 研究的目的是通过一系列综合目标来利用这一机会，这些目标的重点是有效地识别 重要IRD基因的致病性变异。拟议的研究旨在改变目前的研究 范式-分析IRD基因中发现的少量DNA变异-到一个范式 其中预先产生了关于医学上重要的IRD基因中的变体的大量数据。 因此，拟议的研究测试了这样一种假设，即经验性的、基于细胞的测定可以有效地用于 并准确识别人类中哪些DNA变异是致病性的并导致IRD， 良性多态性在目标1中，我们收集和表征了一个全面的潜在的 致病性氨基酸的变化，在一个重要的显性IRD基因，视紫红质。此Aim测试的扩展 这些突变中的哪一个适合于用小分子进行分子伴侣治疗。在Aim 2中，这些技术 被修饰以表征一系列潜在致病性氨基酸变化， 一个重要的隐性IRD基因RPE 65。进一步假设，将测定结果与人的测定结果进行比较， 表型数据将定义对应于人类致病结果的适当数值范围。观看 总之，这些目标提供了一种途径，可以产生一种开放的资源，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关于IRD中VUS的更高保真信息， IRD。它利用了一个重要的机会，对目标的调查可以直接产生 医学上可操作的信息，导致向其他无法治疗的患者提供治疗。

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.