Mechanisms of PIDDosome Signaling, a p53-Independent Apoptotic Response to DNA Damage

PIDDosome 信号传导机制，一种不依赖于 p53 的 DNA 损伤凋亡反应

• 批准号: 10153709
• 负责人: Samuel Sidi
• 金额: $ 34.69万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-05 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10153709
• 关键词: Apoptosis; Apoptotic; BCL2 gene; Binding; Binding Proteins; Biochemical; Bypass; CASP2 gene; CASP8 gene; CASP9 gene; CHEK1 gene; Cancer Etiology; Cancer cell line; Caspase; Cell Nucleolus; Cells; Cessation of life; Chemotherapy and/or radiation; Client; Clinical; Collaborations; Complex; Cues; DNA; DNA Crosslinking Agent; DNA Damage; DNA Interstrand Crosslinking; DNA Repair; DNA Repair Gene; DNA biosynthesis; Data; Death Domain; Diagnostic; Dissection; Drug Targeting; Embryo; Event; FANCD2 protein; Failure; Fanconi anemia protein; Genetic; Goals; Head; Human; Impairment; Induction of Apoptosis; Ionizing radiation; Kinetochores; Knowledge; Lesion; MDM2 gene; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Mediating; Memorial Sloan-Kettering Cancer Center; Mind; Mitochondria; Mitosis; Molecular; Mus; Mutation; NPM1 gene; Nonhomologous DNA End Joining; Nucleolar Proteins; Organelles; Pathway interactions; Phosphorylation; Physiological; Positioning Attribute; Proteins; Proteomics; Radiation; Radiation Tolerance; Radiation therapy; Resistance; Role; Scaffolding Protein; Signal Transduction; Site; Stress; TP53 gene; Tertiary Protein Structure; Time; Topoisomerase Inhibitors; Tumor Suppression; Tumor Suppressor Proteins; Vertebrates; Xenograft Model; Zebrafish; anti-cancer; ataxia telangiectasia mutated protein; chemotherapy; crosslink; druggable target; experimental study; functional genomics; inhibitor/antagonist; insight; metaplastic cell transformation; mutant; neoplastic cell; novel; novel diagnostics; nucleophosmin; prevent; radiation resistance; radiation response; receptor; recruit; repaired; replication stress; response; scaffold; therapy resistant; tool; tumor; tumorigenesis

Project Summary Mutations in p53 and attendant apoptotic pathways impair tumor cell responses to radiation and chemotherapy in many human malignancies. Combining genetics, functional genomics and proteomics in mammalian cells and zebrafish embryos these past 6 years, we identified a novel apoptotic pathway that bypasses p53-dependent pathways altogether via activation of the PIDDosome (PIDD-RAIDD- caspase-2) complex (Sidi et al., Cell 2008; Ando et al., Mol Cell 2012; Thompson et al., Mol Cell 2015; Ando et al., J Cell Biol 2017). Unlike the mitochondrial apoptosome (cytc-Apaf1-caspase-9) and death receptor complex (FAS-FADD-caspase-8), the PIDDosome does not require p53 for activation or function. PIDDosome assembly can be activated by inhibiting its negative regulator, Chk1 kinase. As such, Chk1 inhibitors restore radiosensitivity in p53 mutant zebrafish embryos, MEF, and human cancer cell lines. The PIDDosome is also responsive to DNA damaging chemotherapies such as topoisomerase inhibitors. Altogether, the PIDDosome pathway defines both a novel apoptotic axis and a promising targeted strategy for overcoming treatment resistance in cancer. However, our molecular understanding of the PIDDosome remains very limited. To expand our knowledge of the pathway and identify novel diagnostic tools and drug targets therein, this proposal will focus on the mechanisms by which DNA damage triggers PIDDosome assembly in vertebrate cells. Thus far, we have shown that DNA damage triggers PIDDosome formation via (i) ATM/ATR-mediated phosphorylation of PIDD, which enables RAIDD recruitment to the platform (Mol Cell 2012); and (ii) the binding of PIDD to nucleophosmin (NPM1), which provides a scaffold for PIDDosome assembly (JCB 2017). In Aim 1, we will elucidate the mechanism by which a newly identified PIDD interactor, the DNA repair protein FANCI, recruits PIDD to DNA crosslinks and enables its phosphorylation by ATM at these lesions. Notably, these experiments may identify FANCI as the first biochemically described “repair/death” switch in vertebrates. In Aim 2, we will elucidate the mechanism by which NPM1 and two newly identified nucleolar PIDD-binding proteins, NOLC1 and NCL, coordinately orchestrate PIDDosome formation in response to IR. These experiments may ultimately outline the major apoptotic branch in the nucleolar DNA damage response. Finally, using xenograft models of intrinsic tumor radioresistance (Liu et al., Nat Cell Biol, accepted in principle), we will assess for the first time the potential of PIDDosome targeting as a strategy to overcome radioresistance in TP53 mutant cancers. Altogether, these studies integrate the PIDDosome in the cellular responses to DNA repair failure, replication stress and nucleolar stress. Our proposal is thus ideally positioned to reveal the role of the PIDDosome in cancer etiology, one of the most hotly debated questions in the field of apoptosis.

项目摘要 p53突变和伴随的凋亡途径损害肿瘤细胞对辐射的反应， 化疗在许多人类恶性肿瘤中的应用。结合遗传学、功能基因组学和蛋白质组学 在过去的6年中，我们在哺乳动物细胞和斑马鱼胚胎中发现了一种新的凋亡途径， 通过激活PIDDosome（PIDD-RAIDD-1）完全绕过p53依赖性通路。 caspase-2）复合物（Sidi等人，Cell 2008; Ando等人，Mol Cell 2012; Thompson等人，Mol Cell 2015; Ando等人，J Cell Biol 2017）。不像线粒体线粒体（cytc-Apaf 1-caspase-9）和死亡 受体复合物（FAS-FADD-胱天蛋白酶-8），PIDDosome不需要p53来激活，或 功能PIDDosome组装可以通过抑制其负调节因子Chk 1激酶来激活。作为 这样，Chk 1抑制剂恢复了p53突变斑马鱼胚胎、MEF和人类癌症的放射敏感性 细胞系PIDDosome也对DNA损伤化疗有反应， 拓扑异构酶抑制剂。总之，PIDDosome途径定义了一种新的凋亡轴， 这是一种有希望的克服癌症治疗耐药性的靶向策略。 然而，我们对PIDDosome的分子理解仍然非常有限。扩大我们 该途径的知识，并确定新的诊断工具和药物靶点，这一建议将 专注于脊椎动物细胞中DNA损伤触发PIDDosome组装的机制。 到目前为止，我们已经证明DNA损伤通过（i）ATM/ATR介导的DNA损伤触发PIDDosome形成。 PIDD的磷酸化，其使得RAIDD能够募集到平台（Mol Cell 2012）;和（ii）PIDD的磷酸化，其使得RAIDD能够募集到平台（Mol Cell 2012）。 PIDD与核磷蛋白（NPM 1）的结合，其为PIDDosome组装（JCB）提供支架 2017年）。在目标1中，我们将阐明一种新发现的PIDD相互作用物DNA 修复蛋白FANCI，招募PIDD的DNA交联，并使其磷酸化的ATM在这些 病变值得注意的是，这些实验可以将FANCI鉴定为第一个在生物化学上描述的FANCI。 脊椎动物的“修复/死亡”开关。在目标2中，我们将阐明NPM 1和2 新鉴定的核仁PIDD结合蛋白NOLC 1和NCL协调协调 这些实验可能最终概述了主要的凋亡细胞因子。 核仁DNA损伤反应中的分支。最后，使用内源性肿瘤的异种移植模型， 辐射抗性（Liu等人，Nat Cell Biol，原则上接受），我们将首次评估 PIDDosome靶向作为克服TP 53突变型癌症中的放射抗性的策略的潜力。 总之，这些研究将PIDDosome整合到DNA修复的细胞反应中 失败、复制应激和核仁应激。因此，我们的建议是理想的定位，以揭示作用， PIDDosome在肿瘤病因学中的作用是细胞凋亡领域中最热门的问题之一。