DNA Damage Checkpoint Recovery and Cancer

DNA 损伤检查点恢复与癌症

基本信息

批准号：
8634078
负责人：
Aimin Peng
金额：
$ 30.4万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
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：
2013-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8634078
关键词：
Accounting Address Apoptosis Binding Biochemical Biological Assay Bypass Cancer Patient Cancer cell line Cell Cycle Checkpoint Cell Cycle Progression Cell Death Cell Proliferation Cell-Free System Cells Chemosensitization Cisplatin DNA Damage DNA Repair DNA damage checkpoint Data Event Experimental Models Goals Health Heart In Vitro Investigation Malignant Neoplasms Mitotic Modeling Molecular Mus Oncogenic Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Process Protein Dephosphorylation Protein phosphatase Proteins Radiation Radiation therapy Recovery Recurrence Recurrent Malignant Neoplasm Regulation Role Signal Transduction Site System Testing Therapeutic Tumor Suppression Tumor Suppressor Proteins Up-Regulation Work Xenopus cancer cell cancer therapy chemotherapy egg improved keratinocyte killings neoplastic cell new therapeutic target novel novel therapeutics prevent public health relevance response therapeutic target tool tumor tumor progression tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Cell fate after DNA damage is determined by the cellular DNA damage response (DDR) mechanism that encompasses DNA repair, cell cycle checkpoints, and programmed cell death. Although extensive efforts were dedicated to revealing the activation network of the DDR, little is known about how the cell deactivates the DDR and resumes cell cycle progression, a process termed checkpoint recovery. Importantly, emerging evidence suggests that cancer cells may utilize the recovery mechanism to bypass the DDR during tumorigenesis, and that checkpoint recovery from radio- or chemotherapy causes chemoresistance and tumor recurrence, suggesting checkpoint recovery as an ideal target to improve cancer therapy and patient survival. The long-term goal of this project is to delineate the mechanism of checkpoint recovery and to investigate the implication of checkpoint recovery for cancer progression and therapeutics. We recently characterized the first in vitro system to study checkpoint recovery, and used this system to identify Greatwall (Gwl) kinase as an essential regulator of checkpoint recovery. Interestingly, our preliminary studies discovered novel phosphatases-dependent regulation of checkpoint recovery kinases; our results suggested coordinated activation of Gwl and Plk1, and characterized Gwl phosphorylation and interaction as early events of checkpoint recovery. Moreover, preliminary studies in cancer cell lines and mouse tumor models suggested Gwl as an effective target to enhance chemotherapy and block tumor recurrence. In this proposal we will address fundamental questions regarding the regulatory mechanism of Gwl in checkpoint recovery and its role in cancer therapy and tumor recurrence. Aim 1 will investigate how Gwl and Plk1 are regulated by DNA damage. Our study will take advantage of the cell-free system and a site-specific phosphatase assay to investigate DNA damage-induced, and phosphatase-dependent regulation of Gwl and Plk1. Aim 2 will ask how Gwl and Plk1 re-activate from checkpoint arrest. We will characterize novel Gwl/Plk1 interaction and phosphorylation as potential molecular switches that initiate recovery. In Aim 3, we will characterize how Gwl up-regulation promotes cell recovery from cisplatin treatment and thereby facilitating tumor recurrence. We will then directly evaluate new therapeutic ideas in which targeting Gwl or its initial activation during checkpoint recovery leads to more effective and specific cancer therapy.

描述（由申请人提供）：DNA 损伤后的细胞命运由细胞 DNA 损伤反应 (DDR) 机制决定，该机制包括 DNA 修复、细胞周期检查点和程序性细胞死亡。尽管人们付出了大量的努力来揭示 DDR 的激活网络，但人们对细胞如何停用 DDR 并恢复细胞周期进程（这一过程称为检查点恢复）知之甚少。重要的是，新出现的证据表明，癌细胞在肿瘤发生过程中可能利用恢复机制绕过 DDR，并且放疗或化疗的检查点恢复会导致化疗耐药和肿瘤复发，这表明检查点恢复是改善癌症治疗和患者生存的理想目标。该项目的长期目标是描述检查点恢复的机制并研究检查点恢复对癌症进展和治疗的影响。我们最近表征了第一个用于研究检查点恢复的体外系统，并使用该系统将 Greatwall (Gwl) 激酶识别为检查点恢复的重要调节因子。有趣的是，我们的初步研究发现了检查点恢复激酶的新型磷酸酶依赖性调节；我们的结果表明 Gwl 和 Plk1 协调激活，并将 Gwl 磷酸化和相互作用描述为检查点恢复的早期事件。此外，对癌细胞系和小鼠肿瘤模型的初步研究表明，Gwl 是增强化疗和阻止肿瘤复发的有效靶点。在本提案中，我们将解决有关 Gwl 在检查点恢复中的调节机制及其在癌症治疗和肿瘤复发中的作用的基本问题。目标 1 将研究 Gwl 和 Plk1 如何受 DNA 损伤调节。我们的研究将利用无细胞系统和位点特异性磷酸酶测定来研究 DNA 损伤诱导的 Gwl 和 Plk1 的磷酸酶依赖性调节。目标 2 将询问 Gwl 和 Plk1 如何从检查站逮捕中重新激活。我们将新的 Gwl/Plk1 相互作用和磷酸化描述为启动恢复的潜在分子开关。在目标 3 中，我们将描述 Gwl 上调如何促进顺铂治疗的细胞恢复，从而促进肿瘤复发。然后，我们将直接评估新的治疗理念，其中靶向 Gwl 或其在检查点恢复期间的初始激活导致以获得更有效和特异性的癌症治疗。