Functional Profiling of Human Disease Targets

项目总结 我们在这项建议中的总体目标是从功能上分析与一组 已经获得了大量未表征的遗传变异的复杂疾病模型。 随着了解多个个体的完整基因的前景和日益复杂的 测量表型的方法，生物医学现在可以探索基因与表型的关系 机械化的细节。在确定基因类型时需要解决的一个根本问题是 变异与表型有直接关系。我们的前提是，仅有序列是不够的。现在需要的是 更好地理解基因分型的功能性和机械性分子后果的颠覆性转变 不同之处。我们对这个具有挑战性的问题的解决方案是研究复杂的大分子网络，或者 由大量相互作用的基因和基因产物在细胞内和细胞内形成的“相互作用”。 这些网络的扰动是遗传变异的结果。在表征基因型别时- 通过相互作用组网络方法，基因变异可以导致 完全基因敲除，被建模为移除网络中的一个节点及其所有边，或者，作为替代， 特定于交互作用的扰动，导致移除或加强特定交互作用，建模为 边缘特定的，或“边缘”的扰动。我们认为，为了更好地理解基因-表型 关系，“edgotype”的特征应该是系统地建立节点移除的状态 而不是每个生物物理相互作用的边缘扰动。 这些策略将应用于一小部分复杂的临床表型，因为它们表现出 广泛的遗传异质性、多效性和表型重叠。这四种临床表型(Usher 综合征；视网膜色素变性；先天性巨结肠症；Bardet-Biedl综合征)也得到了足够的研究 已知大量的突变能够在足够的深度下进行EDGING分析以产生 信息丰富的疾病网络。对这四种临床表型的研究应相应地提供基础 对基因型-表型关系的洞察，DNA序列变异对特定生物学的影响 功能、疾病模块和疾病分类。 我们的具体目标是： I)为所选的一组临床表型生成深度和健壮的交互作用组网络图， 二)生成基因产品之间受干扰的物理和生化相互作用的边缘模式图 与选定的一组临床表型有关， 三)通过计算利用EDGOGING数据来获得对基因-表型的机械性分子洞察 选定的一组临床表型的关系。

PROJECT SUMMARY Our overall goal in this proposal is to functionally analyze mutations in human genes associated with a set of model complex disorders for which a large number of uncharacterized genetic variants have been obtained. With the prospect of knowing the complete genotype of multiple individuals and with increasingly sophisticated ways of measuring phenotypes, biomedicine can now explore genotype-phenotype relationships in mechanistic detail. A fundamental issue to be resolved in the characterization of genotypes is how genetic variation directly relates to phenotype. Our premise is that sequence alone is not sufficient. What is needed is a disruptive shift to better understand the functional and mechanistic molecular consequences of genotypic differences. Our solution to this challenging problem is to investigate the complex macromolecular networks, or "interactomes", formed by large numbers of interacting genes and gene products inside cells and to the perturbations of these networks that occur as a consequence of genetic variation. In characterizing genotype- to-phenotype relationships via an interactome network approach, genotypic variation can lead to either a complete gene knockout, modeled as removal of a node and all of its edges in the network, or alternatively, as interaction-specific perturbation, leading to the removal or strengthening of specific interactions, modeled as edge-specific, or "edgetic" perturbations. We propose that to better understand genotype-phenotype relationships, "edgotypes" should be characterized by systematically establishing the state of node removal versus edgetic perturbations for every biophysical interaction. These strategies will be applied to a small set of complex clinical phenotypes chosen because they exhibit extensive genetic heterogeneity, pleiotropy and phenotypic overlap. These four clinical phenotypes (Usher syndrome; retinitis pigmentosa; Hirschsprung disease; Bardet-Biedl syndrome) have also been studied enough that ample numbers of mutations are known to enable edgotyping profiling at sufficient depth to generate informative disease networks. Study of these four clinical phenotypes should accordingly provide fundamental insights into genotype-phenotype relationships, the impact of DNA sequence variants on specific biological functions, disease modules, and disease classification. Our specific aims are to: i) Generate deep and robust interactome network maps for the selected set of clinical phenotypes, ii) Generate edgotypic maps of perturbed physical and biochemical interactions amongst gene products implicated in the selected set of clinical phenotypes, iii) Exploit edgotyping data computationally to derive mechanistic molecular insights into genotype-phenotype relationships for the selected set of clinical phenotypes.