Multi-organism platform for functional analysis of Undiagnosed Diseases Network (UDN) variants

项目摘要 这项申请提出了华盛顿大学圣路易斯医学院(WUSM)的模式生物 筛查核心(WuMOSC)作为NIH未诊断疾病网络(UDN)第二阶段的关键资产。这个 WuMOSC将评估UDN临床站点确定的遗传变异的致病性 未诊断的参与者通过利用两个成熟的动物模式生物资源核心：C. 雅致和斑马鱼。该方法利用了这两个模型系统的实验优势， 同时避免了个别型号的限制。线虫允许极其快速和经济高效的变种 斑马鱼能够评估在线虫中不保守的基因和变异体。一个 已经组建了经验丰富的领导团队，利用合作研究环境， WUSM，包括构成拟议UDN阶段的卓越WUSM临床合作伙伴的专业知识 WUSM临床现场应用。结合使用成熟、尖端和新颖的生物信息学方法 在仔细考虑每种模式生物的优点和局限性后，仔细评估 已经确定了确定指定变种的推定致病性并确定它们的优先顺序的计划 通过资源核进行实验评估。资源核心正在使用高效和先进的遗传 靶向技术，包括CRISPR/Cas9将人类变体敲入到一个同源基因中， 在适当的情况下进行功能下降和功能丧失的分析，并通过过度表达来评估功能丧失的抢救。 或者是为了获得功能收益。遗传模型的表型分析将以模式生物中的信息为指导 数据库和文献，以及病人的症状。然后，这些信息将被应用于生物体- 特定的表型管道。UDN临床站点、指导委员会和协调委员会将 通过世界海关组织的活动，经常收到关于世界海关组织的计划和成果的信息 管理核心，它还将向更广泛的NIH和其他机构传播获得的模式生物专业知识 研究社区。正如过去四年所表明的那样，WuMOSC将继续推进 通过生物信息学和实验评估致病潜力对普遍糖尿病肾病的诊断作用 疾病特定表型背景下的变异。

Project Summary This application proposes the Washington University in St. Louis School of Medicine (WUSM) Model Organism Screening Core (wuMOSC) as a key asset for Phase II of the NIH Undiagnosed Diseases Network (UDN). The wuMOSC will evaluate the pathogenicity of genetic variants identified by UDN Clinical Sites in otherwise undiagnosed participants by leveraging the two well-established animal model organisms Resource Cores: C. elegans and zebrafish. This approach capitalizes on the experimental advantages of these two model systems, while avoiding the limitations of individual models. C. elegans allows for extremely rapid and cost-effective variant evaluation, while zebrafish enables assessment of genes and variants that are not conserved in C. elegans. An experienced Leadership Team has been assembled that harnesses the collaborative research environment at WUSM, including the expertise of the superb WUSM clinical partners that constitute the proposed UDN Phase II WUSM Clinical Site application. Using proven, cutting-edge, and novel bioinformatic approaches, combined with thoughtful consideration of the advantages and limitations of each model organism, a careful assessment plan has been defined for determining the putative pathogenicity of nominated variants and prioritizing them for experimental evaluation by the Resource Cores. The Resource Cores are using efficient and advanced genetic targeting techniques, including CRISPR/Cas9 to knock-in the human variant into an orthologous gene, knock- down and loss-of-function assays where appropriate, and overexpression to assess rescue of loss-of-function or for gain-of-function. Phenotypic analysis of the genetic models will be guided by information in model organism databases and the literature, as well as patient symptoms. This information will then be applied to the organism- specific phenotyping pipelines. The UDN Clinical Sites, Steering Committee, and Coordinating Committee will receive frequent communications regarding plans and results of the wuMOSC, through the activity of the Administrative Core, which will also disseminate acquired model organism expertise to the wider NIH and other research communities. As demonstrated over the past four years, the wuMOSC will continue to advance the diagnostic efforts of the UDN by bioinformatically and experimentally evaluating the potential of disease-causing variants in the context of disease-specific phenotypes.