Chromatin maintenance and sister chromatid differentiation

染色质维持和姐妹染色单体分化

• 批准号: 8535709
• 负责人: PETER M LANSDORP
• 金额: $ 22.26万
• 依托单位: BRITISH COLUMBIA CANCER AGENCY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8535709
• 关键词: Address; Area; Biology; Cell Fate Control; Cell Therapy; Cell division; Cell model; Cells; Centrosome; Chromatin; Chromosomes; DNA; DNA Sequence; DNA biosynthesis; Development; Disease; Elements; Epigenetic Process; Gene Expression; Generations; In Vitro; Inherited; Kinetochores; Lead; Maintenance; Malignant Neoplasms; Microtubules; Mitosis; Modeling; Molecular; Mothers; Mutation; Organism; Research; Sister; Sister Chromatid; Sorting - Cell Movement; Techniques; Testing; Variant; base; cancer stem cell; cell type; daughter cell; imprint; in vivo; induced pluripotent stem cell; novel; public health relevance; research study; segregation; self-renewal; therapy development

DESCRIPTION (provided by applicant): Following replication of parental DNA template strands, sister chromatids are expected to have exactly the same DNA sequence except for mutations resulting from DNA replication itself. How epigenetic marks are transmitted to the next cell generation during cell division is less clear. Epigenetic marks that are not lost prior to mitosis are either distributed randomly between sister chromatids or assigned to a specific sister chromatid e.g. depending on whether the parental DNA template strand was replicated by either leading or the lagging strand DNA replication. Alternatively, sister chromatids could be imprinted in a strand-specific manner. Chromatin differences between sister chromatids could be functionally insignificant or lead to "stochastic" differences in the expression of genes between daughter cells. Epigenetic differences between sister chromatids could also be involved in cell fate regulation if microtubules originating from "mother' centrosomes were to prefer kinetochores present on just one of the two sister chromatids of specific chromosomes. Until recently it was not possible to reliably identify and distinguish sister chromatids. However, we recently found that DNA template strand sequences can be reliably used to identify sister chromatids. These novel techniques will be used to study the possibility of epigenetic differences between sister chromatids and their functional implications. The proposed studies have three specific aims: 1. Study gene expression in relation to specific sister chromatids that were inherited in single cells. 2. Study chromatin marks on sorted sister chromatids containing either "Watson" or "Crick" DNA template strand sequences. 3. Follow the segregation of sister chromatids from specific chromosomes into specific cell types in vitro and in vivo. The proposed studies towards these complementary specific aims will clarify whether chromatin differences between sister chromatids exist and if so, whether such differences are relevant to stochastic variation in gene expression between daughter cells and cell fate decisions and the development in multicellular organisms. Importantly, the proposed studies will answer a question which has not been addressed before: does it matter which sister chromatids are inherited by a given daughter cell? If the answer is affirmative, the proposed studies and experimental approaches will pave the way for further studies of an entirely novel field of research: chromatin replication and sister chromatid differentiation. The implications of this type of research are hard to predict but range from the identification of novel targets for therapy and the development of more effective cell therapies starting from (induced) pluripotent stem cells to more effective strategies that target self-renewal in normal and malignant stem cells.

描述(申请人提供)：复制亲本DNA模板链后，姐妹染色单体预期具有完全相同的DNA序列，但DNA复制本身导致的突变除外。表观遗传标记是如何在细胞分裂期间传递给下一代细胞的，目前还不太清楚。在有丝分裂前没有丢失的表观遗传标记要么随机分布在姐妹染色单体之间，要么分配给特定的姐妹染色单体，例如取决于亲本DNA模板链是通过领先的DNA复制还是通过滞后的DNA复制复制的。或者，姐妹染色单体可以以链特异的方式印记。姐妹染色单体之间的染色质差异在功能上可能微不足道，或者导致子代细胞之间基因表达的“随机”差异。姐妹染色单体之间的表观遗传差异也可能参与细胞命运的调控，如果起源于母亲的中心体的微管倾向于只出现在特定染色体的两个姐妹染色单体中的一个上的动点。直到最近，还不可能可靠地识别和区分姐妹染色单体。然而，我们最近发现，DNA模板链序列可以可靠地用于识别姐妹染色单体。这些新技术将被用来研究姐妹染色单体之间表观遗传差异的可能性及其功能含义。拟议的研究有三个具体目标：1.研究与单细胞遗传的特定姐妹染色单体相关的基因表达。2.研究含有“Watson”或“Crick”DNA模板链序列的姐妹染色单体上的染色质标记。3.遵循特定染色体姐妹染色单体在体外和体内分离到特定细胞类型的情况。针对这些互补的特定目标的拟议研究将澄清姐妹染色单体之间是否存在染色质差异，如果存在，这种差异是否与子细胞之间基因表达的随机变化和细胞命运的决定以及多细胞生物体的发育有关。重要的是，拟议的研究将回答一个以前从未解决过的问题：给定子细胞遗传哪些姐妹染色单体重要吗？如果答案是肯定的，所提出的研究和实验方法将为进一步研究一个全新的研究领域铺平道路：染色质复制和姐妹染色单体分化。这类研究的意义很难预测，但范围从识别新的治疗靶点，从(诱导的)多能干细胞开始开发更有效的细胞疗法，到更有效的针对正常和恶性干细胞自我更新的策略。

项目成果

Epigenetic differences between sister chromatids?

• DOI: 10.1111/j.1749-6632.2012.06505.x
• 发表时间: 2012-01-01
• 期刊: HEMATOPOIETIC STEM CELLS VIII
• 作者: Lansdorp, Peter M.;Falconer, Ester;Naumann, Ulrike
• 通讯作者: Naumann, Ulrike

DNA template strand sequencing of single-cells maps genomic rearrangements at high resolution.

• DOI: 10.1038/nmeth.2206
• 发表时间: 2012-11
• 期刊: NATURE METHODS
• 影响因子: 48
• 作者: Falconer, Ester;Hills, Mark;Naumann, Ulrike;Poon, Steven S. S.;Chavez, Elizabeth A.;Sanders, Ashley D.;Zhao, Yongjun;Hirst, Martin;Lansdorp, Peter M.
• 通讯作者: Lansdorp, Peter M.

Chromosome orientation fluorescence in situ hybridization to study sister chromatid segregation in vivo.

• DOI: 10.1038/nprot.2010.102
• 发表时间: 2010-07
• 期刊: Nature protocols
• 影响因子: 14.8

Telomere length measurement-caveats and a critical assessment of the available technologies and tools.

• DOI: 10.1016/j.mrfmmm.2011.04.003
• 发表时间: 2012-02-01
• 期刊: MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS
• 影响因子: 2.3
• 作者: Aubert, Geraldine;Hills, Mark;Lansdorp, Peter M.
• 通讯作者: Lansdorp, Peter M.

Collapse of telomere homeostasis in hematopoietic cells caused by heterozygous mutations in telomerase genes.

• DOI: 10.1371/journal.pgen.1002696
• 发表时间: 2012
• 期刊: PLoS genetics
• 影响因子: 4.5
• 作者: Aubert G;Baerlocher GM;Vulto I;Poon SS;Lansdorp PM
• 通讯作者: Lansdorp PM