Variant Validation Core

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.