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.

项目概要/摘要内源性DNA损伤对DNA聚合酶运动的干扰，环境和化疗药物既是人类癌症和衰老的原因，也是一个常见的问题许多抗癌药物的作用机制。这种与基因毒素相关的复制应激也是一种很好的公认的 ATR（共济失调毛细血管扩张突变和 rad3 相关）蛋白激酶激活剂，其作用在细胞 DNA 损伤期间调节 DNA 复制和细胞周期相变的关键作用响应（DDR）。 ATR 的小分子抑制剂已成为改善疗效的潜在佐剂。常见癌症化疗方案的有效性。不幸的是，我们对 ATR 作用的理解 DDR 可能偏向于涉及 DNA 合成和细胞分裂的细胞过程，因为通常用于研究 ATR 功能的主动复制细胞的模型系统。然而，使用非通过体外复制培养细胞和离体人体皮肤组织外植体，我们的初步数据揭示了 ATR 激活的新模式与转录应激密切相关，特别是 XPB 亚基多功能蛋白 TFIIH（转录因子 II-H）。此外，我们还发现，与此形成鲜明对比的是 ATR 激酶抑制对复制细胞的 DNA 损伤敏感作用，ATR 抑制在非复制细胞中的作用相反，复制细胞可以保护非复制细胞免受多种 DNA 损伤剂的致命影响。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复制人类细胞中发挥作用，并确定 ATR 抑制剂是否能提供治疗益处暴露于 DNA 损伤剂的人体组织的重要细胞群。本研究的中心假设提议认为，非复制细胞中 ATR 激酶激活机制在以下方面表现出独特的特性：与复制细胞相比，转录相关的 ATR 激酶信号传导有很大不同 DNA 损伤对细胞和组织命运的影响。这项拟议研究的理由是，它将更全面地了解 ATR 激酶如何影响细胞和组织对 DNA 的反应对人体有损害作用，并可能导致使用 ATR 激酶抑制剂来限制某些 DNA 的毒性有害化合物。我们的假设将通过实现以下三个具体目标来检验：1）定义暴露于非复制静止和分化的人类细胞中 ATR 激酶激活的机制遗传毒性应激； 2) 描述非 ATR 激酶抑制的积极和消极后果体外复制细胞； 3) 验证非复制细胞中 ATR 激酶激活和功能的模式人类和小鼠皮肤组织的离体和体内。我们的方法是创新的，因为它将调查人类细胞和组织中 DNA 损伤反应的未探索和生理相关方面。这拟议的研究意义重大，因为它将为 ATR- 的调节提供新的机制见解依赖性 DNA 损伤信号传导可能为人类细胞和组织提供治疗益处。