Functional phenotyping of human genetic variation

人类遗传变异的功能表型

• 批准号: 9001359
• 负责人: RAYMOND J MONNAT
• 金额: $ 38.63万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9001359
• 关键词: Address; Biochemical; Biological Assay; Cell Survival; Cells; Clone Cells; Computer Simulation; DNA; DNA Damage; Data; Disease; Effectiveness; Fanconi Anemia pathway; Fanconi anemia protein; Fanconi' s Anemia; Gene Family; Gene Mutation; Genes; Genetic Determinism; Genetic Variation; Genome; Genome Stability; Genomics; Genotype; Goals; Health; Hereditary Disease; Human; Human Biology; Human Genetics; Link; Mass Spectrum Analysis; Missense Mutation; Modeling; Modification; Mutation; Outcome; Pathway interactions; Phenotype; Play; Proteins; Research; System; Targeted Research; The Cancer Genome Atlas; Time; Variant; Work; base; clinically relevant; crosslink; disorder risk; gene function; genetic variant; improved; innovation; interest; loss of function; member; molecular phenotype; mutant; novel strategies; population based; prediction algorithm; protein expression; protein function; response; tool; variant of unknown significance

DESCRIPTION (provided by applicant): The goal of the proposed research is to develop new tools and a systematic approach to capture and experimentally determine the functional consequences (i.e., the phenotype) of human mutations in high-throughput fashion. This work directly addresses the growing challenge of determining mutation functional consequences, especially for the large number of variants of unknown functional significance (VUS's). We will analyze missense mutations in the 15-member Fanconi anemia (or FANC) gene family that plays key roles in genomic stability and the response to DNA damage, as well as in disease risk and therapy. This gene family provides a biologically interesting and clinically relevant target fo this research. Significance: Genome projects are identifying ever larger numbers of mutations of unknown functional significance (VUS's). The proposed research will establish a facile way to rapidly and systematically determine the functional consequences of large numbers (dozens to hundreds) of mutations in parallel in an isogenic, human cell-based assay system using well-defined, clinically relevant endpoints. Approach: We are integrating computational and new experimental tools to determine in a human cell-based assay system the functional phenotype of all missense mutations in the human Fanconi anemia (FANC) genes. Our Aims are: Aim 1: Capture and in silico analysis of all human Fanconi anemia/FANC gene mutations. Aim 2: High-throughput human cell phenotyping of all FANC gene missense mutations. Aim 3: Integrated, data-driven modeling of FANC protein function in response to DNA damage. Innovation: The proposed research has four innovative features: 1. global, ascertainment bias-free capture of all genetic variation in the FANC gene families; 2. systematic mutation functional prediction and rank-scoring using six different prediction algorithms; 3. a new, high-throughput cell-based functional phenotyping assay that is quantitative, and built on clinically relevant cellular endpoints; and 4. the use of mutation phenotyping and molecular data to build an integrated dynamic model of Fanconi protein function in response to DNA damage. Anticipated outcomes: The proposed research will establish a systematic approach for determining the functional phenotype of human mutations including the growing number of VUS's. Results of this work should substantially improve our understanding of Fanconi gene function, while providing a general approach and new tools to enable other human biology and genetic disease research.

描述（由申请人提供）：拟议研究的目的是开发新工具和一种系统的方法，以高通量方式捕获和实验确定人类突变的功能后果（即表型）。这项工作直接解决了确定突变功能后果的日益增长的挑战，尤其是对于大量未知功能意义的变体（VUS）的挑战。我们将分析15名成员的Fanconi贫血（或Fanc）基因家族中的错义突变，该基因在基因组稳定性以及对DNA损伤以及疾病风险和治疗中起关键作用。该基因家族为这项研究提供了一个在生物学上有趣且与临床相关的目标。意义：基因组项目正在识别出更多功能意义的突变（VUS）。拟议的研究将建立一种迅速，系统地确定使用临床上相关的临床相关终点在同基因的，基于人类细胞的基于人类细胞的测定系统中并行确定大量突变（数十至数百个突变）的功能后果。方法：我们正在整合计算和新的实验工具，以在人类细胞的测定系统中确定人类法科尼贫血（FANC）基因中所有错义突变的功能表型。我们的目的是：目标1：捕获和在所有人类狂欢节贫血/狂欢基因突变中捕获和分析中。 AIM 2：所有狂欢基因错义突变的高通量人类细胞表型。 AIM 3：响应DNA损伤的综合，数据驱动的模型。创新：拟议的研究具有四个创新特征：1。全球，确定性的无偏见捕获粉丝基因家族中所有遗传变异的无偏见； 2。使用六种不同预测算法的系统突变功能预测和排名得分； 3。一种新型的，高通量的基于细胞的功能表型测定，它是定量的，并建立在临床相关的细胞终点上；和4。使用突变表型和分子数据来响应DNA损伤构建Fanconi蛋白功能的综合动态模型。预期的结果：拟议的研究将建立一种系统的方法来确定人类突变的功能表型，包括越来越多的VUS。这项工作的结果应大大提高我们对Fanconi基因功能的理解，同时提供一种通用方法和新工具，以实现其他人类生物学和遗传疾病研究。