Greatwall in replication stress/DNA damage responses and oral cancer resistance

长城在复制应激/DNA损伤反应和口腔癌抵抗中的作用

• 批准号: 10545740
• 负责人: Aimin Peng
• 金额: $ 36.34万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-05 至 2023-10-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10545740
• 关键词: ATM activation; Back; Biochemical; Biology; Cell Cycle; Cell Proliferation; Cells; Chemicals; Cisplatin; Clinical; Collaborations; DNA Damage; DNA biosynthesis; DNA replication fork; Data; Databases; Dentistry; Development; Diagnosis; Distant Metastasis; Drug Sensitization; Drug Targeting; Drug resistance; Event; Fluorouracil; Goals; Head and neck structure; Hour; Human Papillomavirus; Investigation; Laboratories; Ligase; Malignant Epithelial Cell; Malignant Neoplasms; Mediating; Mitosis; Modeling; Mouth Neoplasms; Mus; Neoplasm Metastasis; Oncologist; Operative Surgical Procedures; Oropharyngeal; Pathologist; Pathway interactions; Patients; Pharmaceutical Preparations; Pharyngeal structure; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Poly(ADP-ribose) Polymerase Inhibitor; Process; Prognosis; Protein Phosphatase 2A Regulatory Subunit PR53; Proteins; Proteolysis; Proteomics; Publishing; Radiation; Recovery; Recurrent tumor; Regimen; Regulation; Residual Neoplasm; Resistance; Role; Signal Transduction; Squamous cell carcinoma; Therapeutic; Time; Ubiquitination; United States; Up-Regulation; biological adaptation to stress; cancer cell; cancer drug resistance; cancer therapy; cancer type; chemoradiation; chemotherapeutic agent; chemotherapy; clinical development; college; cytotoxicity; effective therapy; hydroxyurea; improved; in vivo; inhibitor; insight; malignant mouth neoplasm; mouth squamous cell carcinoma; neoplastic cell; new therapeutic target; novel therapeutics; recruit; repaired; replication factor A; replication stress; response; small molecule inhibitor; spatiotemporal; targeted cancer therapy; therapeutic target; therapy resistant; tumor; tumor progression; tumor xenograft

Oral cancer, including cancers of the mouth and the back of the throat, is the sixth most common cancer worldwide. In the United States, approximately 50,000 new oral cancer cases are diagnosed each year. First- line treatments for oral cancer typically include surgery and radiation, with chemotherapy added to decrease the possibility of metastasis, to eliminate residual tumor cells after surgery, to enhance the efficacy of radiation (chemoradiation), and for patients with confirmed distant metastasis. Radiation and oral cancer chemotherapeutics confer cytotoxicity largely by disrupting DNA replication to induce DNA damage. Unfortunately, the prognosis of oral cancer, particularly HPV(-) cases, remains relatively poor, calling for a better understanding of how cells respond to replication stress and DNA damage, and accordingly, developing more effective treatment options and combinations to overcome drug resistance. In the current project, we characterize a new role of Greatwall (Gwl) kinase in the replication stress and DNA damage responses. Gwl was frequently upregulated in HPV(-) oral cancer, in correlation with cancer progression, tumor recurrence, and poor patient survival. Gwl promoted the recovery and resistance of oral cancer cells to drugs that induce replication stress and DNA damage. Gwl depletion or inhibition sensitized the drug responses in oral cancer cells and mouse tumor models. Building on these findings, we hypothesize that Gwl mediates the cellular responses to replication stress and DNA damage, and is therefore a potent target for oral cancer therapy. We will uncover detailed mechanisms underlying the function and regulation of Gwl in the replication stress and DNA damage responses; we will also establish the crucial proof-of-principle for the development of Gwl targeting in cancer treatment. In Aim 1, we will delineate how Gwl is recruited to stalled replication forks via its interaction with replication protein A (RPA) to regulate a phosphatase-mediated response to replication stress. This study will shed new light on cancer progression and treatment, given that replication stress is a hallmark of cancer, and that anti-replication drugs are commonly used in cancer therapy. In Aim 2, we will reveal a new mechanism that leads to Gwl stabilization and accumulation after replication stress and DNA damage, potentially as a key event that initiates cell recovery and confers tumor resistance. Upregulation of Gwl is mediated directly by DNA damage signaling, suggesting a self-engaged “timer” mechanism that initiates cell recovery and treatment resistance. Finally, guided by our mechanistic investigations, we will explore in Aim 3 therapeutic targeting of Gwl, using unique small molecule inhibitors which interfere with either Gwl kinase activation or its interaction with RPA. Both patient-derived oral tumor xenograft and orthotopic syngeneic oral tumor models will be utilized to comprehensively evaluate the therapeutic potential of Gwl inhibition. Together, this project will lead to a deeper understanding of the cellular responses to replication stress and DNA damage, and characterize a new drug target to improve oral cancer therapy.

口腔癌，包括口腔癌和咽喉后部癌，是第六大常见癌症 全世界。在美国，每年诊断出大约 50,000 例新的口腔癌病例。第一的- 口腔癌的一线治疗通常包括手术和放疗，并添加化疗以减少 转移的可能性，消除手术后残留的肿瘤细胞，增强放疗的疗效 （放化疗），以及已确诊远处转移的患者。放射与口腔癌 化疗药物主要通过破坏 DNA 复制以诱导 DNA 损伤来赋予细胞毒性。 不幸的是，口腔癌，特别是 HPV(-) 病例的预后仍然相对较差，需要采取措施 更好地了解细胞如何应对复制压力和 DNA 损伤，并相应地开发 更有效的治疗方案和组合来克服耐药性。在当前的项目中，我们 描述了 Greatwall (Gwl) 激酶在复制应激和 DNA 损伤反应中的新作用。古尔 在 HPV(-) 口腔癌中经常上调，与癌症进展、肿瘤复发、 以及患者生存率低下。 Gwl促进口腔癌细胞的恢复和对诱导药物的抵抗 复制压力和 DNA 损伤。 Gwl 消耗或抑制使口腔癌的药物反应敏感 细胞和小鼠肿瘤模型。基于这些发现，我们假设 Gwl 介导细胞 对复制应激和 DNA 损伤的反应，因此是口腔癌治疗的有效靶点。我们 将揭示 Gwl 在复制应激和调节中的功能和调节的详细机制 DNA损伤反应；我们还将为 Gwl 的发展建立关键的原理验证 癌症治疗中的靶向。在目标 1 中，我们将描述 Gwl 如何通过其 与复制蛋白 A (RPA) 相互作用，调节磷酸酶介导的复制应激反应。 鉴于复制压力是一个标志，这项研究将为癌症进展和治疗提供新的线索 抗复制药物常用于癌症治疗。在目标 2 中，我们将揭示一个新的 复制应激和 DNA 损伤后导致 Gwl 稳定和积累的机制， 可能是启动细胞恢复并赋予肿瘤抵抗力的关键事件。 Gwl 的上调是 由 DNA 损伤信号直接介导，表明启动细胞的自参与“计时器”机制 恢复和治疗抵抗力。最后，在我们的机制研究的指导下，我们将在目标 3 中进行探索 使用干扰 Gwl 激酶的独特小分子抑制剂以 Gwl 为治疗靶点 激活或其与 RPA 的相互作用。患者来源的口腔肿瘤异种移植和原位同基因口腔肿瘤 肿瘤模型将用于综合评价Gwl抑制的治疗潜力。一起， 该项目将导致人们更深入地了解细胞对复制压力和 DNA 的反应 损伤，并表征新的药物靶点以改善口腔癌的治疗。