DNA damage response kinase signaling in non-replicating human cells and tissues

非复制人类细胞和组织中的 DNA 损伤反应激酶信号传导

基本信息

批准号：
10091482
负责人：
Michael George Kemp
金额：
$ 29.6万
依托单位：
WRIGHT STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10091482
关键词：
ATM function ATR gene Adjuvant Aging Antineoplastic Agents Biological Models Cancer Etiology Cell Cycle Cell Cycle Progression Cell Death Cell Differentiation process Cell Survival Cell division Cell physiology Cells Chemotherapy-Oncologic Procedure Cultured Cells DNA DNA Damage DNA biosynthesis DNA lesion DNA replication fork DNA-Directed DNA Polymerase DNA-Directed RNA Polymerase Data Disease susceptibility ERCC3 gene Effectiveness Epidermis Eukaryota Event Exhibits Exposure to Foundations Genetic Transcription Genotoxic Stress Goals Health Holoenzymes Homeostasis Human Human body In Vitro KRP protein Lead Life Link Malignant Neoplasms Mission Mitotic Modeling Movement Mus Mutagenesis Mutagens Organ Outcome Pathway interactions Phase Phase Transition Phosphotransferases Physiological Play Population Prevention Process Property Protein Kinase Proteins Public Health Publishing RNA Polymerase II Research Risk Role Signal Transduction Skin Skin Tissue Source Stress Testing Therapeutic Tissues Toxic effect Ultraviolet B Radiation United States National Institutes of Health Work ataxia telangiectasia mutated protein base biological adaptation to stress cancer therapy carcinogenesis cell type chemotherapeutic agent environmental agent genotoxicity homologous recombination human tissue improved in vivo inhibitor/antagonist innovation insight irradiation kinase inhibitor novel programs replication stress response small molecule inhibitor stem cell differentiation stressor transcription factor S-II transcription factor TFIIH

项目摘要

PROJECT SUMMARY/ABSTRACT The interference of DNA polymerase movement by DNA damage induced by endogenous, environmental, and chemotherapeutic agents is both a cause of cancer and aging in humans and a common mechanism of action of many anti-cancer drugs. This genotoxin-associated replication stress is also a well- recognized activator of the ATR (ataxia telangiectasia-mutated and rad3-related) protein kinase, which plays critical roles in regulating DNA replication and cell cycle phase transitions during the cellular DNA damage response (DDR). Small molecule inhibitors of ATR have emerged as potential adjuvants to improve the effectiveness of common cancer chemotherapy regimens. Unfortunately, our understanding of the role of ATR in the DDR may be biased towards cellular processes involving DNA synthesis and cell division because of the model systems of actively replicating cells that are typically used to study ATR function. However, using non- replicating cultured cells in vitro and human skin tissue explants ex vivo, our preliminary data have revealed a novel mode of ATR activation that is closely linked with transcription stress and specifically with the XPB subunit of the multi-functional protein TFIIH (transcription factor II-H). Moreover, we have found that in striking contrast to the DNA damage-sensitizing effects of ATR kinase inhibition on replicating cells, ATR inhibition in non- replicating cells instead protects non-replicating cells from the lethal effects of several DNA damaging agents. The objective of this proposal is to therefore more clearly define the mechanisms of ATR kinase activation and function in non-replicating human cells and to determine whether ATR inhibitors provide therapeutic benefit to important cell populations of human tissues exposed to DNA damaging agents. The central hypothesis of this proposal is that the mechanism of ATR kinase activation in non-replicating cells exhibits unique properties in comparison to replicating cells and that transcription-associated ATR kinase signaling has profoundly different effects on cell and tissue fate in response to DNA damage. The rationale for this proposed research is that it will provide a more complete understanding of how the ATR kinase impacts cellular and tissue responses to DNA damaging agents in humans and may lead to the use of ATR kinase inhibitors to limit the toxicity of certain DNA damaging compounds. Our hypothesis will be tested by carrying out the following three specific aims: 1) Define the mechanism of ATR kinase activation in non-replicating quiescent and differentiated human cells exposed to genotoxic stress; 2) Characterize the positive and negative consequences of ATR kinase inhibition in non- replicating cells in vitro; and 3) Validate the modes of ATR kinase activation and function in non-replicating cells of human and mouse skin tissue ex vivo and in vivo. Our approach is innovative because it will investigate an unexplored and physiologically relevant aspect of the DNA damage response in human cells and tissues. The proposed research is significant because it will provide novel mechanistic insights into how modulation of ATR- dependent DNA damage signaling may provide therapeutic benefits in human cells and tissues.

项目摘要/摘要 DNA聚合酶通过内生引起的DNA损伤的干扰，环境和化学治疗剂既是癌症的原因，又是人类衰老的原因，也是常见的许多抗癌药物的作用机理。这种基因毒素相关的复制应力也是一种很好的公认的ATR（共济失调telangiectia-mutated and rad3相关）蛋白激酶的活化剂在细胞DNA损伤期间调节DNA复制和细胞周期过渡中的关键作用响应（DDR）。 ATR的小分子抑制剂已成为潜在的佐剂，以改善普通癌症化疗方案的有效性。不幸的是，我们对ATR的作用的理解在DDR中，可能会偏向涉及DNA合成和细胞分裂的细胞过程模型的积极复制细胞的系统，通常用于研究ATR功能。但是，使用非 - 在体外复制培养的细胞和人类皮肤组织外植体，我们的初步数据显示了一个新型ATR激活模式与转录应力紧密相关，特别是XPB亚基多功能蛋白TFIIH（转录因子II-H）的此外，我们发现在引人注目的对比中对于ATR激酶抑制对复制细胞的DNA损伤敏化作用，非ATR抑制复制细胞改为保护非复制细胞免受几种DNA损伤剂的致命作用。因此，该提案的目的是更清楚地定义ATR激酶激活的机制和在非重复的人类细胞中起作用，并确定ATR抑制剂是否为治疗益处提供了治疗益处暴露于DNA破坏剂的人体组织的重要细胞群。中心假设建议是非复制细胞中ATR激酶激活的机制在与复制细胞进行比较，该转录相关的ATR激酶信号具有深远的不同对DNA损伤响应细胞和组织命运的影响。这项拟议研究的理由是它将对ATR激酶如何影响细胞和组织对DNA的反应有更完整的了解人类破坏药物，可能导致使用ATR激酶抑制剂来限制某些DNA的毒性有害化合物。我们的假设将通过执行以下三个特定目的来检验：1）定义 ATR激酶激活在非复制静止和分化的人类细胞中的机制遗传毒性应激； 2）表征非ATR激酶抑制在非 - 体外复制细胞； 3）验证非复制细胞中ATR激酶激活和功能的模式人体和小鼠皮肤组织的体内和体内。我们的方法是创新的，因为它将调查人类细胞和组织中DNA损伤反应的未开发和生理相关的方面。这拟议的研究很重要，因为它将提供新的机械洞察力，以了解如何调节ATR- 依赖的DNA损伤信号传导可能会在人类细胞和组织中提供治疗益处。