Genomic Fate Maps

基因组命运图谱

• 批准号: 7760668
• 负责人: MARSHALL S. HORWITZ
• 金额: $ 32.82万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7760668
• 关键词: Achievement; Address; Affect; Anatomy; Animal Model; Back; Biology; Birth Records; Caenorhabditis elegans; Cataloging; Catalogs; Cell Count; Cell Line; Cells; Central Vein; Cessation of life; Chemicals; Collaborations; Computer-Assisted Three-Dimensional Imaging; DNA biosynthesis; Development; Developmental Biology; Devices; Dissection; Drops; Dyes; Embryo; Ethylnitrosourea; Frequencies; Gene Expression; Genetic Markers; Genetic Recombination; Genome; Genomics; Goals; Hand; Hepatic; Hepatocyte; Individual; Inherited; Injury; Knowledge; Label; Length; Letters; Life; Liver; Mammalian Cell; Mammals; Maps; Methods; Microdissection; Microscope; Microtome - medical device; Mismatch Repair; Molecular; Mus; Mutagenesis; Mutation; NCI Center for Cancer Research; Organ; Organism; Phylogenetic Analysis; Population; Positioning Attribute; Recording of previous events; Repetitive Sequence; Resolution; Sampling; Somatic Mutation; Stem cells; Stream; System; Techniques; Technology; Time; Tissues; Trees; Validation; base; distraction; in vivo; laser capture microdissection; novel; progenitor; research study; tissue regeneration; zygote

DESCRIPTION (provided by applicant): Recent, surprising, and controversial discoveries have challenged conventional concepts regarding the origins and plasticity of stem cells, and their contributions to tissue regeneration, and highlight just how little is known about mammalian development in comparison to simpler model organisms. In the case of the transparent worm, C. elegans, Sulston and colleagues used a microscope to record the birth and death of every cell during its life, and the compilation of this fate maps represents a milestone achievement of developmental biology. Determining a fate map for mammals or other higher organisms is more complicated, because they are opaque, take a long time to mature, and have a tremendous number of cells. Consequently, fate mapping experiments have relied on tagging a progenitor cell with a dye or genetic marker in order to later identify its descendants. This approach, however, extracts little information, because it only reveals that a population of cells, all inheriting the same label, share a common ancestor but tells nothing of how each cell is related to one another. To avoid that, as well as the technical limitations of current methods for mapping cell fate, we propose a new strategy for retrospectively deriving cell fate maps by using phylogenetics to infer the order in which somatic mutations have arisen in the genomes of individual cells during the development of multicellular organisms. DNA replication introduces mutations, particularly at repetitive sequences, every time a cell divides. In Preliminary Studies we demonstrate that cataloging the frequent mutations affecting the length of polyguanine tracts allows for deducing the history of mouse cells either passaged in culture or sampled from the various tissues of a single individual. Toward an ultimate goal of constructing a high resolution mammalian cell fate map, we plan three Specific Aims: 1. Compare phylogenetic fate maps to conventional fate maps derived from genetically marked mice. 2. Advance the technology of this approach. 3. Successively build a liver cell fate map in order to address controversies regarding the clonal, embryonic, and anatomic origins of hepatic stem cells during development and in disease.A goal of biology is to understand how a single cell divides and gives rise to all the different tissues and organs in the body. Fate maps describe how cells within an individual are related to one another and can be likened to a tree of life, in which the origin of a particular cell can be traced back through its progenitors to the fertilized egg. We propose a new method for exploring developmental biology by constructing fate maps based on distinctive mutations inevitably accumulating in the genome each time a cell divides. We will initially focus our studies on how the liver develops and responds to injury.

描述（由申请人提供）：最近，令人惊讶和有争议的发现挑战了有关干细胞的起源和可塑性及其对组织再生的贡献的常规概念，并且强调了与简单模型生物体相比，对哺乳动物发育的了解知之甚少。在透明蠕虫的情况下，秀丽隐杆线虫，硫磺和同事使用显微镜记录了每个细胞在其生命中的出生和死亡，并且该命运图的汇编代表了发展生物学的里程碑。确定哺乳动物或其他较高生物体的命运图更为复杂，因为它们是不透明的，需要很长时间才能成熟，并且具有大量细胞。因此，命运映射实验依赖于用染料或遗传标记的祖细胞标记祖细胞，以便稍后确定其后代。但是，这种方法提取了很少的信息，因为它仅揭示了一个遗传同一标签的细胞群共享一个共同的祖先，但对每个细胞之间的相互关系却一无所知。为了避免这种情况，以及当前绘制细胞命运方法的技术局限性，我们提出了一种回顾性得出细胞命运图的新策略，该策略通过使用系统发育学来推断多细胞生物发展过程中单个细胞基因组中体细胞突变产生的顺序。 DNA复制会引入突变，尤其是在重复序列时，每次细胞分裂时。在初步研究中，我们证明，对影响聚圭氨酸长度长度的频繁突变进行分类，可以推断出在培养物中转移或从单个个体的各种组织中取样的小鼠细胞的史。为了构建高分辨率的哺乳动物细胞命运图的最终目标，我们计划了三个特定目的：1。将系统发育命运图与源自遗传标记的小鼠得出的常规命运图进行比较。 2.推进这种方法的技术。 3。为了解决有关在发育和疾病中肝干细胞的克隆，胚胎和解剖学起源的争议。生物学的目标是了解单个细胞分裂并引起人体中所有不同的组织和器官。命运图描述了一个人内部的细胞如何相互关系，并可以比作生命树，其中特定细胞的起源可以通过其祖细胞追溯到受精卵。我们提出了一种新方法，通过基于独特的突变构建命运图，每次细胞分裂时都不可避免地积累了基因组中的命运图。我们最初将重点关注肝脏如何发展和应对伤害的研究。