Dissecting the genetic determinants of aneuploidy's effect on cellular proliferat

剖析非整倍体对细胞增殖影响的遗传决定因素

基本信息

批准号：
8313461
负责人：
Anna Brosius Sunshine
金额：
$ 3.5万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2014-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): An abnormal karyotype, including both segmental and whole chromosomal aneuploidies, is a hallmark of cancer. Many tumors are aneuploidy and there is evidence that aneuploidy may be a driver of tumorigenesis [27]. Despite these close ties to cancer, aneuploidy is paradoxically associated with a cell cycle delay and decreased growth rates [25, 29]. Aneuploidy results in the coordinated copy-number change of tens to thousands of genes. This genetic complexity has made it difficult to understand how aneuploidy might promote tumorigenesis and cellular proliferation. Saccharomyces cerevisiae is a well-studied model organism with many elements of its cellular biology conserved with mammalian cells. The study of complex genetic problems is simplified in S. cerevisiae due to the relative simplicity of the yeast genome and by the many available strain collections. In a process with many parallels to tumorigenesis, our laboratory has isolated aneuploid clones of S. cerevisiae from a subset of 24 independent long-term evolution experiments [7]. Just as aneuploidies frequently represented in a population of cancer cells are hypothesized to be associated with a proliferative advantage [17], similarly, in Aim 1 we plan to identify high-frequency aneuploid clones isolated from these evolution experiments and in Aim 2 determine their proliferative advantage. Population-level analysis of the evolution experiments will be carried out using array comparative genomic hybridization (aCGH) of population DNA while the relative proliferative advantage of different strains will be determined by direct competition experiments. In Aim 3 we will exploit bar-coded yeast collections spanning more than 90% of the protein coding genome [10, 30] and the novel high-throughput sequencing technology Bar-seq [21] to determine how the coordinated copy- number change of multiple genes within an aneuploid region combines to affect cellular proliferation. We hope that the rigorous genetic analysis of aneuploidy proposed here will help us better understand the role of aneuploidy in tumorigenesis and cancer progression. PUBLIC HEALTH RELEVANCE: Aneuploidy is a genetic abnormality where a cell has an abnormal number of chromosomes. Cancer is a disease characterized by unbridled cellular growth and many cancer cells themselves are aneuploid [reviewed in 28]. In this proposal we will use yeast to identify the genetic basis for aneuploidy's effects on cellular growth.

描述（由申请人提供）：异常的核型，包括分段和整个染色体非整倍性，是癌症的标志。许多肿瘤是非整倍性的，有证据表明非整倍性可能是肿瘤发生的驱动器[27]。尽管与癌症有亲密关系，但非整倍性与细胞周期延迟和生长速率降低相关[25，29]。非整倍性导致数十基因的协调拷贝数变化。这种遗传复杂性使难以理解各个非整倍性如何促进肿瘤发生和细胞增殖。酿酒酵母是一种经过良好研究的模型生物，其细胞生物学的许多元素用哺乳动物细胞保存。由于酵母基因组的相对简单性和许多可用的菌株收集，在酿酒酵母中简化了复杂遗传问题的研究。在与肿瘤发生的许多相似之处的过程中，我们的实验室已将酿酒酵母的非整倍体克隆与24个独立的长期进化实验的子集分离出来[7]。正如癌细胞群中经常代表的非整倍性被认为与增殖优势相关[17]，同样，在AIM 1中，我们计划鉴定从这些进化实验中分离出的高频非频型克隆，并且在AIM 2中确定其增殖优势。将使用阵列比较基因组杂交（ACGH）进行进化实验的人群级分析，而种群DNA的相对增殖优势将通过直接竞争实验确定。在AIM 3中，我们将利用跨越超过90％蛋白质编码基因组[10，30]和新型的高通量测序技术Bar-Seq [21]的条形编码的酵母集合[21]来确定如何在Aneuploid区域内的多个基因的拷贝数变化，以影响细胞扩散。我们希望这里提出的对非整倍性的严格遗传分析将有助于我们更好地了解非整倍性在肿瘤发生和癌症进展中的作用。公共卫生相关性：非整倍性是一种遗传异常，其中细胞具有异常数量的染色体。癌症是一种以无限制的细胞生长为特征的疾病，许多癌细胞本身都是非整倍性的[在28中综述]。在此提案中，我们将使用酵母来确定非整倍性对细胞生长的影响的遗传基础。