Variant Validation Core

RES-Core-001(034) 项目摘要 靶向肾脏基因板、全外显子组测序和全基因组测序在慢性肾脏疾病和其他肾脏疾病的遗传学基础研究中的广泛应用和日益增多，使人们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基因诊断，在许多情况下改善了临床护理。此外，有了基因诊断，患者可以考虑通知他们的亲属，他们可能也会受到影响。但已经发现的许多单核苷酸变种既不是明显致病的，也不是明显良性的；这些被称为不确定意义的变种(VUS)。建议的变异验证核心(VVC)的目的是调查在肾脏疾病或肾脏发育障碍患者中发现的VU的潜在致病性。WC将帮助使用者有效和经济地确定在患者身上发现的不能被归类为致病或良性的DNA/RNA/蛋白质变异的致病潜力。VVC将用于硅胶和细胞培养方法，通过比较野生型和变异体来分析蛋白质功能、RNA剪接和基因表达。此外，CRISPR/Cas9介导的基因编辑将用于产生携带VU的适当小鼠模型；这些模型将被分析是否患有肾脏疾病或障碍。用户带到VVC的每个VU的实验方法将针对1)潜在涉及的特定基因和蛋白质；以及2)如果VU确实是致病的，可能会发展成的特定疾病或障碍而量身定做。对于临床医生和研究人员来说，VVC将是一个宝贵的国家资源，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的遗传基础的更大研究中找到VUS。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，从而潜在地影响患者及其家人。VVC将与华盛顿大学的其他奥布莱恩核心进行互动，并将与国家奥布莱恩联盟和指导委员会协调，为用户提供最高质量的服务。

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.