Variant Validation Core

Res-Core-001（034） 项目摘要 广泛和越来越多地使用靶向肾脏基因组、全外显子组测序和全基因组测序来研究慢性肾脏疾病和其他肾脏疾病的遗传基础，这使得对单基因疾病的病因学有了更好的理解。这些方法为患者提供了明确的遗传诊断，并在许多情况下改善了临床护理。此外，基因诊断使患者可以考虑通知他们的亲属他们也可能受到影响。但是有许多单核苷酸变异已被发现，既不是明确的致病性，也不是明确的良性;这些被称为变异的不确定意义（VUS）。拟议的变体验证核心（VVC）旨在研究在肾脏疾病或肾脏发育障碍患者中发现的VUS的潜在致病性。WC将帮助用户高效且具有成本效益地确定在患者中发现的DNA/RNA/蛋白质变异体的致病性潜力，这些变异体不能被归类为致病性或良性。VVC将在二氧化硅和细胞培养方法中使用，通过比较野生型与变体来测定蛋白质功能、RNA剪接和基因表达。此外，CRISPR/Cas9介导的基因编辑将用于生成携带VUS的适当小鼠模型;这些模型将用于分析肾脏疾病或病症。针对用户带到VVC的每个VUS的实验方法将针对1）可能涉及的特定基因和蛋白质;以及2）如果VUS确实是致病性的，则可能发展的特定疾病或病症进行定制。VVC将成为临床医生和研究人员的宝贵国家资源，他们将VUS作为常规临床护理的一部分，或在旨在确定各种肾脏疾病遗传基础的大型研究中发现。VVC进行的实验结果将与患者诊断直接相关，并可能为临床护理提供信息，从而可能影响患者及其家属。VVC将与华盛顿大学的其他奥布莱恩核心互动，并与国家奥布莱恩联盟和指导委员会协调，为用户提供最优质的服务。

Res-Core-001 (034) Project Summary The widespread and increasing use of targeted kidney gene panels, whole exome sequencing, and whole genome sequencing to investigate the genetic bases for chronic kidney disease and other kidney disorders has led to a much better understanding of the etiology of monogenic diseases. These approaches have provided patients with clear genetic diagnoses and in many cases have led to improved clinical care. In addition, having a genetic diagnosis has allowed patients to consider informing their relatives that they might also be affected. But there are many single nucleotide variants that have been discovered that are neither clearly pathogenic nor clearly benign; these are termed variants of uncertain significance (VUS). The aim of the proposed Variant Validation Core (VVC) is to investigate VUS discovered in patients with kidney disease or a kidney developmental disorder for potential pathogenicity. The WC will assist users in the efficient and cost-effective determination of the potential for pathogenicity of DNA/RNA/protein variants discovered in patients that cannot be classified as pathogenic or benign. The VVC will use in silica and cell culture approaches to assay protein function, RNA splicing, and gene expression by comparing wild-type to variants. In addition, CRISPR/Cas9- mediated gene editing will be used to generate appropriate mouse models carrying VUS; these will be analyzed for kidney disease or disorders. The experimental approach for each VUS brought to the VVC by a user will be tailored for 1) the particular gene and protein that are potentially involved; and 2) the particular disease or disorder that would likely develop if the VUS is indeed pathogenic. The VVC will be a valuable national resource for both clinicians and researchers who find VUS as part of routine clinical care or in larger research studies aimed at defining the genetic bases for diverse kidney diseases. The results of the experiments carried out by the VVC will be directly relevant to patient diagnosis and may be informative for clinical care, thus potentially impacting both patients and their families. The VVC will interact with the other O'Brien Cores at Washington University and will coordinate with the National O'Brien Consortium and Steering Committee to provide the highest quality services for users.