Effect of anti-S phase agents on human chromosomes

抗S期药物对人类染色体的影响

基本信息

批准号：
8608500
负责人：
Anindya Dutta
金额：
$ 31.8万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-02-01 至 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8608500
关键词：
A549 Address Affect Cell Adhesion Molecules Cell Cycle Cell Line Cell Lineage Cell Proliferation Cell surface Cells ChIP-seq Chromatin Chromosomal Breaks Chromosomal Rearrangement Chromosomal translocation Chromosome Fragility Chromosome Structures Cytarabine DNA DNA biosynthesis DNA lesion DNA repair protein Double Strand Break Repair Doxorubicin Drug toxicity Drug usage Fanconi&apos s Anemia Fetal Hemoglobin Fire - disasters Gene Expression Gene Expression Profile Genes Genetic Transcription Genome Genomics H1299 Hela Cells Hematologic Neoplasms Hematological Disease Human Human Chromosomes Kinetics Ku70 protein Lead Lesion Location MCF7 cell Malignant neoplasm of cervix uteri Malignant neoplasm of lung Mediating Messenger RNA Methods Nature Neighborhoods Pathway interactions Pattern Pharmaceutical Preparations Polycythemia Vera Positioning Attribute Proteins Publishing RNA Polymerase II Recruitment Activity Replication Origin Research S Phase Second Primary Cancers Sequence Analysis Sickle Cell Anemia Site TREX1 gene Techniques Testing Time Study Tumor Suppressor Proteins base cancer therapy cell type density drug testing essential thrombocytopenia gemcitabine hydroxyurea improved inhibitor/antagonist malignant breast neoplasm next generation sequencing novel promoter public health relevance repaired response tumor progression

项目摘要

DESCRIPTION (provided by applicant): Anti-S phase agents like doxorubicin, gemcitabine, cytarabine and hydroxyurea are in extensive use for the treatment of cancers and of hematological diseases such as essential thrombocytopenia, polycythemia vera and sickle cell anemia. All these agents are believed to cause replication fork stalling leading to activation of checkpoint pathways that inhibit cell proliferation. Yet, not much is known about how these inhibitors inhibit the origins of replication, of the sites/DNA lesions responsible for checkpoint activation and of the effect of these lesions on chromosome structure or gene expression. In addition, many of these agents are known to affect gene expression by unknown mechanisms. For example, hydroxyurea has been used in sickle cell anemia because it increases the expression of fetal hemoglobin and decreases the expression of cell-surface adhesion molecules. We have recently published genomic approaches to study time of replication, origins of replication and effects of anti-S phase agents on the replication origins in human cells We have discovered that two anti-S phase agents used for therapy lead to the firing of clustered neo-origins, leading to a high density of stalled replication forks at defined sites in the genome. In this proposal we plan to determine whether this is a general feature of other anti-S phase agents, whether the sites where these clustered neo-origins occur are the same in different cell lines or whether the sites change when cells are exposed to these agents in all parts of the S phase. We will investigate whether the high density of stalled replication forks at clustered neo-origins make these sites of the genome susceptible to double-strand breaks (DSB) and chromosomal rearrangements. We will examine whether the disturbed state of the chromatin due to the high density of clustered neo-origins leads to suppression of gene expression. Finally, we will examine which, where and in what order checkpoint activators and DNA repair proteins are recruited to the stalled replication forks created by anti-S phase agents.

描述（由申请人提供）：抗S期药物如阿霉素、吉西他滨、阿糖胞苷和羟基脲广泛用于治疗癌症和血液疾病，如原发性血小板减少症、真性红细胞增多症和镰状细胞性贫血。所有这些药物均被认为会导致复制叉停滞，从而激活抑制细胞增殖的检查点途径。然而，关于这些抑制剂如何抑制复制起点、负责检查点激活的位点/DNA损伤以及这些损伤对染色体结构或基因表达的影响，人们知之甚少。此外，已知许多这些试剂通过未知机制影响基因表达。例如，羟基脲已被用于治疗镰状细胞性贫血，因为它增加胎儿血红蛋白的表达并减少细胞表面粘附分子的表达。我们最近发表了基因组方法来研究复制时间、复制起点以及抗 S 期药物对人类细胞复制起点的影响我们发现用于治疗的两种抗 S 期药物会导致簇状新细胞的激发-起源，导致基因组中特定位点出现高密度的停滞复制叉。在本提案中，我们计划确定这是否是其他抗S期药物的一般特征，这些聚集的新起源出现的位点在不同的细胞系中是否相同，或者当细胞暴露于这些药物时这些位点是否发生变化在S期的所有部分。我们将研究聚集的新起源处高密度的停滞复制叉是否使基因组的这些位点容易受到双链断裂（DSB）和染色体重排的影响。我们将检查由于高密度聚集的新起源而导致的染色质紊乱状态是否会导致基因表达的抑制。最后，我们将检查哪些检查点激活剂和 DNA 修复蛋白被招募到由抗 S 期药物产生的停滞复制叉、在何处以及以什么顺序。