Screening for modifiers of PKD severity using ENU Mutagenesis

摘要 从对人群和小鼠模型的分析中有大量证据表明， 多囊肾病（PKD）可以通过相互作用的遗传基因座进行修饰。这些位点的鉴定 应该提供深入了解我们的理解的基本病理生物学的囊肿和疾病 进展重要的是，它们可能揭示治疗干预的新途径。我们有 在囊性肾病小鼠模型表征方面的丰富经验，特别是 调查其严重程度的菌株特异性修饰剂。然而，在小鼠中已证实的致病基因的产量 这方面的研究很少。相比之下，我们使用不同的方法非常成功， 新的疾病基因发现，即用化学品乙基亚硝基脲（ENU）诱变。我们有 我最近修改了这种方法，这样我们就可以完全在近交系背景上进行筛选， 用于定位克隆的基因组测序方法。PKD1RC突变体的最新特征 小鼠具有缓慢进展的PKD，对菌株特异性修饰剂敏感，迫使我们提出 我们使用ENU诱变来产生和发现PKD1诱导的囊性肾的修饰物， 疾病为了补充这种表型驱动的方法，我们还将对候选基因座进行分析， 可能改变PKD的严重程度。我们有数据表明，Sonic Hedgehog（SHH）信号在 我们将测试是否在这一途径中的基因缺失会影响疾病的严重程度， PKD1RC小鼠模型。

ABSTRACT There is abundant evidence from the analysis of human populations and mouse models that the severity of Polycystic Kidney Disease (PKD) can be modified by interacting genetic loci. The identification of these loci should provide insight into our understanding of the basic pathobiology of cystogenesis and disease progression. Importantly, they can potentially reveal novel pathways of therapeutic intervention. We have extensive experience in the characterization of a mouse model of cystic kidney disease, and specifically the investigation of strain-specific modifiers of its severity. However, the yield of proven causal genes in mouse studies of this type has been low. In contrast, we have been very successful using a different approach for novel disease gene discovery, namely mutagenesis with the chemical ethyl-nitrosourea (ENU). We have recently modified this method so that we can do our screen entirely on an inbred background, using Whole Genome Sequencing methodology for positional cloning. The recent characterization of the PKD1RC mutant mouse as having slowly progressive PKD, which is sensitive to strain-specific modifiers, compels our proposal that we use ENU mutagenesis for the generation and discovery of modifiers of PKD1-induced cystic kidney disease. To complement this phenotype-driven approach, we will also pursue an analysis of candidate loci that may modify PKD severity. We have data to suggest that Sonic Hedgehog (SHH) signaling plays a role in cystogenesis, and we will test whether the deletion of genes in this pathway affects disease severity in the PKD1RC mouse model.