NIH Director's Pioneer Award

摘要：细胞命运图描述了细胞分裂、迁移和凋亡的顺序如何改变受精卵。 是理解干细胞生物学的基础。然而，它只存在于透明的蠕虫C. 对每一个细胞分裂进行繁琐的显微观察，从而构建了一个完整的 细胞命运图。事实证明，越复杂和不透明的动物就越不容易屈服。然而，DNA复制不可避免地 产生体细胞突变。因此，多细胞生物由马赛克组成，其中大多数细胞 获取独特的基因组，这些基因组有可能描绘出它们的祖先。我们建议建造 使用系统发育学方法被动追溯胚胎关系的哺乳动物细胞命运图 推断出在发育过程中发生突变的顺序。我们发现聚鸟嘌呤 重复DNA序列是特别有用的遗传标记，因为它们在 有丝分裂。为了证明可行性，我们使用系统发育学来重建培养的小鼠的谱系。 基于突变影响聚鸟嘌呤标记长度的NIH3T3成纤维细胞。然后我们雇佣了 从小鼠的单个细胞中进行全基因组扩增以对聚鸟嘌呤标志物进行分型 系统发育学来推断样本组织的发育关系。我们的初步结果 展示了这种方法的潜力，可以追溯到产生一个完整的哺乳动物细胞命运， 原则上，可以描述任何细胞的发育谱系并解决与以下方面相关的悬而未决的问题 干细胞生物学。

Abstract: Cell fate maps describe how the sequence of cell division, migration, and apoptosis transform a zygote into an adult and are fundamental to understanding stem cell biology. Yet, it is only in the transparent worm C. elegans where tedious microscopic observation of each cell division has allowed for construction of a complete cell fate map. More complexand opaqueanimals prove less yielding. DNA replication, however, inevitably generates somatic mutations. Consequently, multicellular organisms comprise mosaics where most cells acquire unique genomes that are potentially capable of delineating their ancestry. We propose to construct mammalian cell fate maps using a phylogenetic approach to passively retrace embryonic relationships by deducing the order in which mutations have arisen during development. We have found that polyguanine repeat DNA sequences are particularly useful genetic markers, because they frequently change length during mitosis. To demonstrate feasibility, we have used phylogenetics to reconstruct the lineage of cultured mouse NIH3T3 fibroblasts based on mutations affecting the length of polyguanine markers. We have then employed whole genome amplification to genotype polyguanine markers in single cells taken from a mouse and used phylogenetics to infer the developmental relationships of the sampled tissues. Our preliminary results demonstrate the potential of this approach for retrospectively producing a complete mammalian cell fate that, in principle, could describe the developmental lineage of any cell and resolve outstanding questions relevant to stem cell biology.