GENETIC DISSECTION OF NATURAL VARIATION IN CALORIC RESTRICTION-INDUCED CELLULAR L

热量限制诱导的细胞 L 自然变异的基因解剖

• 批准号: 8718860
• 负责人: Christopher S Nelson
• 金额: $ 5.15万
• 依托单位: BUCK INSTITUTE FOR RESEARCH ON AGING
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8718860
• 关键词: Age; Aging; Aging-Related Process; Alleles; Behavior; Biochemical; Biological Assay; Biology; Caloric Restriction; Cell Aging; Cell Death; Cell Proliferation; Cells; Chromosome Mapping; Chronic Disease; DNA; DNA Sequence; Data; Diet; Disease; Dissection; Endothelial Cells; Engineering; Environment; Environmental Risk Factor; Future; Gene Expression; Genes; Genetic; Genetic Determinism; Genetic Epistasis; Genetic Polymorphism; Genetic Variation; Genome; Genotype; Glucose; Goals; Growth; Human; Individual; Inherited; Intervention; Laboratory Study; Lead; Life; Light; Link; Longevity; Malignant Neoplasms; Maps; Measurement; Measures; Mediating; Metabolism; Methods; Microfluidics; Modeling; Molecular; Molecular Genetics; Mus; Mutagenesis; Mutation; Myelogenous; Nature; Nutritional; Oak Tree; Pathway interactions; Population; Proteins; Quantitative Trait Loci; Rattus; Regulation; Reporter; Research; Role; Signal Transduction; Starvation; Stem cells; Stress; Surveys; System; Techniques; Testing; Validation; Variant; Vascular Diseases; Yeast Model System; Yeasts; aging gene; base; cell age; fitness; fly; genetic analysis; genetic linkage analysis; genetic variant; longevity gene; meetings; molecular scale; novel; novel diagnostics; novel therapeutics; nutrition; overexpression; promoter; public health relevance; reconstruction; research study; response; tool; trait

DESCRIPTION (provided by applicant): Replication and environmental stresses sustained over a cell's lifetime cause damage and ultimately lead to different types of cell death. Understanding the precise molecular mechanisms by which cells age is a major unsolved problem of modern biology. A comprehensive understanding of cellular aging will shed light on chronic diseases such as myeloid cancer and vascular disease, which result from hematopoetic stem cell aging and endothelial cell aging, respectively. Environmental factors have a significant impact on longevity, and one of the best-documented lifespan-extending perturbations is caloric restriction (CR). Yeast, worms, flies, mice, and rats all live longer when reared on CR diets, via a mechanism that remains poorly understood. Genome-scale screens for determinants of the CR response have not been tractable to date; I propose to meet this challenge by harnessing natural genetic variation in the CR response, using wild yeast isolates as a model. My preliminary studies have revealed robust changes between yeast strains in lifespan during CR, including a wild isolate with a distinctly long lifespan under CR conditions. I hypothesize that th genetic determinants of the lifespan extension, growth behavior, and regulatory profile of wild yeast under CR conditions include novel determinants of the CR response. I will test this hypothesis with the following specific aims: Aim 1- I will map the genetic basis of differences between yeast strains in lifespan under CR conditions. Using a panel of genotyped progeny from a cross between two wild yeast isolates, I will assay the lifespan of each with established microfluidic techniques. Statistical-genetic analyses will identify genes and polymorphisms that reproducibly segregate with the lifespan trait, and the role of these factors in lifespan will be validated in independent molecular-genetic experiments. Aim 2- To understand how CR modulates cell proliferation and fitness, I will measure growth traits under CR conditions in the progeny from the cross used in Aim 1, and I will map genetic loci linked to growth behaviors via statistical-genetic methods. Molecular validation of these screen hits will establish genes that mediate the sensing of and response to CR at the cellular level. Comparison to the results of Aim 1 will enable a test of the degree to which lifespan and growth under CR are under shared genetic control. Aim 3- To investigate at a systems level the state of cells grown under CR, I will transcriptionally profile each strain from the cross used in Aim 1. I will map sequence variants that co-inherit with the expression response to CR, and validate these loci in molecular experiments. The results will identify components and connections of the gene network that regulates cell state in response to the nutritional environment, dovetailing with the first two Aim and allowing a genome-scale molecular dissection of growth and longevity decisions during CR.

描述（由申请人提供）：复制和环境应力在细胞的寿命中持续造成损害，并最终导致不同类型的细胞死亡。了解细胞年龄是现代生物学的主要未解决问题的确切分子机制。对细胞衰老的全面理解将揭示出慢性疾病，例如髓样癌和血管疾病，这分别由血管干细胞衰老和内皮细胞衰老造成。环境因素对寿命有重大影响，而延伸寿命最好的驱动力之一是热量限制（CR）。酵母，蠕虫，苍蝇，小鼠和大鼠在Cr饮食中饲养时都寿命更长，该机制仍然很熟悉。迄今为止，针对CR反应决定因素的基因组规模筛选尚未处理。我建议使用野生酵母分离株作为模型来利用CR反应中的自然遗传变异来应对这一挑战。我的初步研究表明，在CR期间，酵母菌菌株之间的稳健变化，包括在CR条件下具有明显长的寿命的野生分离株。我假设在CR条件下野生酵母的寿命扩展，生长行为和调节性谱的遗传决定因素包括CR反应的新决定性。我将以以下特定目的检验这一假设：目标1-我将在CR条件下生命周期中酵母菌菌株之间差异的遗传基础。我将使用来自两个野生酵母菌株之间的十字架的基因分型后代，我将用已建立的微流体技术测定每个人的寿命。统计遗传学分析将确定与寿命性状可重复分离的基因和多态性，并且这些因素在寿命中的作用将在独立的分子遗传实验中得到验证。目标2-为了了解CR如何调节细胞的增殖和适应性，我将在AIM 1中使用的十字架的后代中测量CR条件下的生长特征，并且我将通过统计基因遗传学方法绘制与生长行为相关的遗传基因座。这些筛选命中的分子验证将建立介导细胞水平上CR感应和反应的基因。与AIM 1的结果进行比较，将对CR下的寿命和生长在共享遗传控制下的寿命和生长程度进行测试。目标3-要在系统级别调查CR下生长的细胞状态，我将 从AIM 1中使用的十字架中的每个应变介绍了每个应变。我将绘制与CR的表达响应共同启用的序列变体，并在分子实验中验证这些基因座。结果将确定基因网络的组成部分和连接，该基因网络对响应营养环境的响应，与前两个目标相吻合，并允许在CR期间对生长和寿命决策的基因组规模剖析。