Discovering the spectrum of natural somatic mosaicism in human skin fibroblasts

 描述（由申请人提供）：新出现的证据表明，与先前的假设相反，并非人体的所有细胞都具有相同的DNA序列，这种现象称为体细胞变异。体细胞变异已被证明会导致癌症和其他几种疾病，但这种现象在人体中的程度仍然未知，部分原因是体细胞突变通常发生在一小部分细胞中，并且可能无法在整个人群中检测到。因此，研究这种现象对于我们了解人体如何运作，是什么原因/使我们容易患上各种疾病，以及我们如何避免和/或治愈它们具有根本的重要性。本文介绍了单细胞基因组分析的计算方法和实验方法的发展，以及如何应用这些方法和手段研究人皮肤成纤维细胞的体细胞嵌合现象。该提案首先建议开发一种基于单细胞克隆扩增的复杂，全面的体细胞变异发现方法。然后，这种方法将被应用于揭示人体皮肤成纤维细胞中的体细胞嵌合现象，并将导致在该组织中的体细胞变异的参考集。该项目的完成将为单细胞测序分析提供新的计算和实验分析方法。这些方法可用于各种单细胞研究（体细胞嵌合、生物体发育等）。并且在诊所中（例如，癌症监测）。此外，该结果将提供对人体基因组变异的更好理解，这最终与以下方面相关：i）理解和预防不同人对不同疾病的易感性; i）估计不同治疗对每个特定人的有效性;以及iii）为个性化医疗奠定基础。

 DESCRIPTION (provided by applicant): Emerging evidence suggests that, contrary to previous assumptions, not all cells of the human body have an identical DNA sequence, a phenomenon known as somatic variation. Somatic variations have been shown to cause cancer and several other diseases, but the extent of this phenomenon in the human body remains unknown, in part because somatic mutations often occur in a small proportion of cells and may not be detected in the whole population. Therefore, studying this phenomenon has fundamental importance to our understanding of how the human body functions, what causes/predisposes us to various diseases, and how we can avoid and/or cure them. This proposal describes the development of computation methods and experimental approaches for the analysis of single cell's genomes, as well as how to apply these methods and approaches to study somatic mosaicism in human skin fibroblasts. The proposal first suggests developing a sophisticated, comprehensive approach for somatic variation discovery based on single cell clonal expansion. Then, this approach will be applied to uncover somatic mosaicism in human skin fibroblasts cells and will result in a reference set of somatic variants in this tissue. The result of completig the project will be new computational and experimental analytical approaches for single cell sequencing analysis. These approaches can be used in various single cell studies (somatic mosacism, organism development, etc.) and in the clinic (e.g., for cancer monitoring). Moreover, the result will provide a better understanding of genome variations in human body, which ultimately is relevant in: i) understanding and preventing the susceptibility of different people t different diseases; i) estimating the effectiveness of different treatments for each particular person; and, iii) creating the foundation for personalized medicine.