Phylogenetic Fate Mapping: Following Cellular Lineages in Embryogenesis and Aging

系统发育命运图谱：追踪胚胎发生和衰老中的细胞谱系

• 批准号: 7274577
• 负责人: Stephen J Salipante
• 金额: $ 3.17万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7274577
• 关键词: Adult; Affect; Age; Aging; Animals; Apoptosis; Biology; Brain; Caenorhabditis elegans; Cell Lineage; Cell division; Cells; Complex; DNA Sequence; DNA biosynthesis; Development; Disease; Embryo; Embryonic Development; Facility Construction Funding Category; Generations; Genetic Markers; Genome; Genotype; Health; Heart; Hepatic; Hepatocyte; Individual; Length; Liver; Mammals; Maps; Methods; Microscopic; Mitosis; Mus; Mutation; Nematoda; Newborn Infant; Organ; Organism; Organogenesis; Pattern; Phylogenetic Analysis; Population Study; Process; Production; Recording of previous events; Role; Series; Somatic Mutation; Statistical Methods; Stem cells; Structure; Technology; Thinking; Time; Tissue Sample; Trees; anticancer research; base; daughter cell; microbial; migration; research study; senescence; zygote

DESCRIPTION (provided by applicant): The charting of cellular lineages, also referred to as cell fate mapping, has diverse applications in the study of embryogenesis and aging, and has led to a more comprehensive understanding of health and disease. However, the only organism for which a complete cell fate map has been constructed is the simple, transparent roundworm C. elegans, for which it is possible to microscopically observe each cell division. Fate mapping in higher organisms has relied on indirect methods where a cell is visually or genetically tagged in order to later identify its descendants, an approach that is more limiting and substantially less informative than direct observation. In mammals, however, new mutations arise with almost every mitosis, implying that most cells acquire unique genomes. The inheritance of such spontaneous somatic mutations by daughter cells can be thought of as a naturally occurring record of cell divisions, in which the order that mutations have arisen during development reflects cells' ancestral lineage relationships. We propose a phylogenetic approach to constructing fate maps, in which we adapt phylogenetic methods developed to study population structure in evolutionary and microbial biology to retrospectively trace lineage relationships based on cells' patterns of somatic mutations. Preliminary studies have shown that polyguanine repeat DNA sequences are valuable as highly mutable genetic markers, frequently changing length during mitosis. We have phylogenetically reconstructed the history of an artificial cell lineage tree of cultured mouse NIH3T3 cells based on such mutations affecting the length of polyguanine markers. We then have employed whole genome amplification to genotype polyguanine markers in single cells taken from an adult mouse and have used phylogenetics to construct a proof-of principle fate map of the sampled tissues. The aims of this project are to refine and validate the technology of "phylogenetic fate mapping", and to produce a series of second generation phylogenetic fate maps of the mouse liver in order to resolve several outstanding controversies regarding the embryonic origins of the liver, and the production of hepatocytes in the aging animal. As a technology, phylogenetic fate mapping has the potential to make broad contributions to cancer research, the study of mammalian embryogenesis, and the ongoing role of stem cells in the aging organism. We hope that the proposed studies will serve as the basis for examining the processes of organogenesis and aging in more heterogeneous and structurally complex organs.

描述（由申请人提供）：细胞谱系的图表，也称为细胞命运映射，在胚胎发生和衰老的研究中具有不同的应用，并且导致对健康和疾病的了解更全面。但是，构建完整的细胞命运图的唯一生物是简单的透明round虫秀丽隐杆线虫，可以从显微镜上观察每个细胞分裂。较高生物体中的命运映射依赖于间接方法，这些方法在视觉或遗传上标记了细胞以稍后识别其后代，这种方法比直接观察更具限制和信息性差。然而，在哺乳动物中，几乎所有有丝分裂都会出现新突变，这意味着大多数细胞都获得了独特的基因组。子细胞对这种自发的体细胞突变的遗传可以被认为是一种天然存在的细胞分裂记录，其中突变在发育过程中产生的顺序反映了细胞的祖先谱系关系。我们提出了一种用于构建命运图的系统发育方法，其中我们适应了为研究进化和微生物生物学中种群结构而开发的系统发育方法，以回顾性地基于细胞的体细胞突变模式追踪谱系关系。初步研究表明，聚鸟氨酸重复的DNA序列是有价值的遗传标记，在有丝分裂过程中经常变化。我们基于影响多圭氨酸标记长度的这种突变，从系统发育中重建了培养的小鼠NIH3T3细胞的人造细胞谱系的历史。然后，我们在从成年小鼠中采集的单个细胞中利用了整个基因组扩增来对基因型的聚鸟氨酸标记，并使用系统发育学来构建采样组织的原理证明。 该项目的目的是完善和验证“系统发育命运映射”的技术，并生成一系列小鼠肝脏的第二代系统发育命运图，以解决有关肝脏胚胎起源的几个杰出争议，以及在衰老动物中产生的肝细胞的产生。 作为一项技术，系统发育命运映射有可能为癌症研究，哺乳动物胚胎发生的研究以及干细胞在衰老生物体中的持续作用做出广泛贡献。我们希望拟议的研究将作为研究更异质和结构复杂的器官中器官发生和衰老过程的基础。