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：
8997926
负责人：
Anindya Dutta
金额：
$ 32.79万
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/8997926
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 Repair Gene DNA biosynthesis DNA lesion DNA replication fork 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 Genomic approach 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 TP53 gene TREX1 gene Techniques Testing Time 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相剂，例如阿霉素，吉西他滨，西南滨和羟基脲，可用于治疗癌症和血液学疾病，例如必需的血小板减少症，多余细胞症，多余细胞症和镰状细胞。据信所有这些药物都会引起复制叉的停滞，从而导致抑制细胞增殖的检查点途径的激活。然而，对于这些抑制剂如何抑制复制的起源，负责检查点激活的部位/DNA病变的起源以及这些病变对染色体结构或基因表达的影响，并不了解。此外，已知许多这些药物通过未知机制影响基因表达。例如，羟基脲已用于镰状细胞贫血中，因为它增加了胎儿血红蛋白的表达并降低了细胞表面粘附分子的表达。我们最近发表了研究复制时间，复制的起源和抗S相剂对人类细胞中复制起源的作用的起源，我们发现，用于治疗的两种抗S相剂导致簇状的新奥尔科蛋白发射，导致固定的基因组中固定的复制叉的高密度。在此提案中，我们计划确定这是否是其他抗S相试剂的一般特征，在不同的细胞系中，这些聚类的新单根蛋白发生的位点是否相同，或者当细胞在S相的所有部分暴露于这些药物时这些位点是否会发生变化。我们将研究聚集的新孤子蛋白处的停滞复制叉的高密度是否会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的新染色蛋白而导致的染色质状态是否导致基因表达的抑制。最后，我们将研究将哪种顺序检查点激活剂和DNA修复蛋白募集到抗S相位剂创建的停滞复制叉中的哪些顺序和DNA修复蛋白。