Metnase, PIKK, and RPA Roles in DNA Damage and Replication Stress Responses

Metnase、PIKK 和 RPA 在 DNA 损伤和复制应激反应中的作用

• 批准号: 9100800
• 负责人: Jac A Nickoloff
• 金额: $ 27.04万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9100800
• 关键词: Apoptosis; Autophagocytosis; Breast; Cancer Etiology; Cancer Patient; Cell Cycle Arrest; Cell Cycle Progression; Cell Death; Cell Line; Cell Survival; Cells; Cessation of life; Chemotherapy-Oncologic Procedure; Chimeric Proteins; Chromosomes; Colon; Complex; DNA Damage; DNA Double Strand Break; DNA Integration; DNA Repair; DNA Repair Pathway; DNA Replication Damage; DNA biosynthesis; DNA lesion; DNA replication fork; DNA-dependent protein kinase; Defect; Development; Double Strand Break Repair; Foundations; Genetic Epistasis; Genome Stability; Genotoxic Stress; Growth; Head and neck structure; Health; Human; Hypersensitivity; Induction of Apoptosis; KRP protein; Lead; Lung; Mediating; Methyltransferase; Mitotic; Necrosis; Neoplasm Metastasis; Nonhomologous DNA End Joining; Normal Cell; Pancreas; Pathway interactions; Phosphatidylinositols; Phosphorylation; Play; Protein Kinase; Protein Methyltransferases; Proteins; RAD9A gene; Radiation therapy; Radio; Recovery; Resistance; Risk; Role; S Phase; Single-Stranded DNA; Site; Stress; System; Testing; Therapeutic; Time; Tumor Cell Line; Tumor Suppression; biological adaptation to stress; cancer genome; cancer therapy; cell type; chemotherapy; clinically relevant; cytotoxicity; design; human DNA damage; improved; inhibitor/antagonist; killings; neoplastic cell; novel; nuclease; prevent; programs; repaired; replication factor A; response; senescence; tumor; tumor progression

DESCRIPTION (provided by applicant): Most cancer patients receive radio- and/or chemotherapy that causes DNA damage, which blocks DNA replication. Normal and tumor cells respond to DNA damage and associated replication stress by activating DNA repair, cell cycle arrest (checkpoint) systems, and when damage is severe, programmed death pathways, collectively termed the DNA damage response (DDR). DDR proteins play crucial roles in tumor suppression and genome stabilization (cancer etiology) as well as tumor response to radio- and chemotherapy (cancer treatment). DDR pathways determine cell fates in response to DNA damage, including cell survival, genome stability, and cell death/permanent growth arrest via apoptosis, autophagy, necrosis, senescence, or mitotic catastrophe. Cells are particularly vulnerable to DNA damage during S phase because most DNA lesions stall replication forks, causing replication stress. This proposal focuses on several proteins with roles in DNA repair, checkpoint activation, and recovery from replication stress. Metnase and DNA-PK were both initially characterized for their roles in DNA double-strand break (DSB) repair by non-homologous end joining (NHEJ). Recent studies demonstrate that Metnase and DNA-PK (along with replication protein A (RPA), ATM/ATR, Chk1, and others) also function in checkpoint activation and replication stress recovery. The replication checkpoint prevents new origin firing and stabilizes stalled replication forks to prevent fork collapse, allowing time for repair and for restart. Persistent replication stress can lead to fork collapse, producing one-ended DSBs marked by phosphorylated H2AX (γ-H2AX). RPA accumulates on single-stranded DNA at stalled forks and the RPA32 subunit is phosphorylated at multiple sites by phosphoinositide 3-kinase-related protein kinases (PIKKs) DNA-PK, ATM and ATR, leading to Chk1 activation and replication arrest. Metnase also regulates Chk1 activation and replication arrest. Our central hypothesis is that Metnase, DNA-PK, and RPA operate within the DDR to influence cell fate after genotoxic stress, including cell survival, genome stability, and death pathway activation. We will determine roles of Metnase (Aim 1) and PIKK phosphorylation of RPA (Aim 2) in replication stress responses including replication arrest, fork restart, genome stability, cell survival and cell death by apoptosis. In Aim 3 we will define epistatic relationships between Metnase and PIKK/RPA pathways, and test novel combinations of replication stress agents plus DDR inhibitors to enhance killing of breast, lung, pancreatic, colon, head and neck, and leukemic tumor cells. A better understanding of how DDR factors regulate cell fate decisions will drive the development of novel cancer therapies to improve local tumor control, and reduce the risk of therapy-induced tumor progression and secondary tumor induction.

描述（由应用提供）：大多数癌症患者接受了导致DNA损伤的放射和/或化学疗法，这阻断了DNA复制。正常和肿瘤细胞通过激活DNA修复，细胞周期停滞（检查点）系统以及损伤严重的，编程的死亡途径，对DNA损伤和相关复制应力做出反应，统称为DNA损伤响应（DDR）。 DDR蛋白在肿瘤抑制和基因组稳定（癌症病因）以及对放射和化学疗法的肿瘤反应（癌症治疗）中起着至关重要的作用（癌症治疗）。 DDR途径确定了对DNA损伤的响应细胞命运，包括细胞存活，基因组稳定性以及细胞死亡/永久生长停滞，通过凋亡，自噬，坏死，感应或有丝分裂灾难。细胞在S期特别容易受到DNA损伤的影响，因为大多数DNA病变失速复制叉会导致复制应力。该提案的重点是几种在DNA修复，检查点激活和复制应力中恢复中具有作用的蛋白质。最初，MetNase和DNA-PK都以非同理学端连接（NHEJ）在DNA双链断裂（DSB）修复中的DNA双链破裂（DSB）修复中的作用来表征。最近的研究表明，MetNase和DNA-PK（以及复制蛋白A（RPA），ATM/ATR，CHK1等）在检查点激活和复制应力恢复中也起作用。复制检查点可防止新的起源射击并稳定停滞的复制叉，以防止叉子倒塌，从而使时间进行维修和重新启动。持续的复制应力会导致分叉崩溃，产生以磷酸化为标志的一端DSB H2AX（γ-H2AX）。 RPA在失速的叉子处积聚在单链DNA上，RPA32亚基通过磷酸肌醇3-激酶相关的蛋白激酶（Pikks）DNA-PK，ATM和ATT在多个位点磷酸化，导致CHK1激活和复制停滞。 Metnase还调节CHK1激活和复制停滞。我们的核心假设是，Metnase，DNA-PK和RPA在DDR内运行到目标3中，我们将定义Metnase与PIKK/PIKK/RPA途径之间的认识关系，并测试复制应激剂以及DDR抑制剂的新型组合，以增强乳房，肺，pancreatic，pancreatic，Coleatic，Coleatic，Colean，Headem，Heartem，Headem and tumor，tumor，tumor，tumor，tumiac，tumiac，tumiac，tumor。更好地了解DDR因素如何调节细胞脂肪的决策将推动新型癌症疗法的发展，以改善局部肿瘤控制，并降低治疗诱导的肿瘤进展和继发性肿瘤诱导的风险。

项目成果

FOXF1 mediates mesenchymal stem cell fusion-induced reprogramming of lung cancer cells.

• DOI: 10.18632/oncotarget.2413
• 发表时间: 2014-10-15
• 期刊: Oncotarget
• 作者: Wei HJ;Nickoloff JA;Chen WH;Liu HY;Lo WC;Chang YT;Yang PC;Wu CW;Williams DF;Gelovani JG;Deng WP
• 通讯作者: Deng WP

Improving cancer therapy by combining cell biological, physical, and molecular targeting strategies.

通过结合细胞生物学、物理和分子靶向策略来改善癌症治疗。

• DOI: 10.3978/j.issn.1000-9604.2013.01.06
• 发表时间: 2013
• 期刊: Chinese journal of cancer research = Chung-kuo yen cheng yen chiu
• 作者: Nickoloff,JacA

Assaying DNA double-strand break induction and repair as fast as a speeding comet.

• DOI: 10.4161/cc.24667
• 发表时间: 2013-05-01
• 期刊: Cell cycle (Georgetown, Tex.)
• 作者: Nickoloff JA

Roles of homologous recombination in response to ionizing radiation-induced DNA damage.

• DOI: 10.1080/09553002.2021.1956001
• 发表时间: 2023
• 期刊: International journal of radiation biology
• 影响因子: 2.6

The DNA repair component Metnase regulates Chk1 stability.

DNA 修复成分 Metnase 调节 Chk1 稳定性。

• DOI: 10.1186/1747-1028-9-1
• 发表时间: 2014
• 期刊: Cell division
• 影响因子: 2.3
• 作者: Williamson,ElizabethA;Wu,Yuehan;Singh,Sudha;Byrne,Michael;Wray,Justin;Lee,Suk-Hee;Nickoloff,JacA;Hromas,Robert
• 通讯作者: Hromas,Robert