Defining NEMO interactions to selectively inhibit NF-kB activation by DNA damage

定义 NEMO 相互作用以选择性抑制 DNA 损伤引起的 NF-kB 激活

• 批准号: 8526749
• 负责人: Shawn Siavash Jackson
• 金额: $ 3.69万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-18 至 2016-03-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8526749
• 关键词: Address; Adverse effects; Amino Acids; Antineoplastic Agents; Apical; Attenuated; Bacterial Infections; Behavior; Binding; Biochemical; Cell Line; Cell Survival; Cellular Stress; Clinical; DNA; DNA Damage; Data; Defect; Development; Disease Progression; Effectiveness; Genes; Genetic; Genetic Screening; Infection; Inflammation; Ionizing radiation; Maps; Mediating; Modeling; Molecular; Mutagens; N-terminal; NF-kappa B; Natural Immunity; Pathway interactions; Peptides; Pharmaceutical Preparations; Physiological Processes; Proteins; Radiation-Sensitizing Agents; Reagent; Research; Resistance; Role; Signal Pathway; Signal Transduction; Site; Solutions; Stream; TRAF6 gene; Testing; Therapeutic; Viral; Work; adaptive immunity; anticancer treatment; base; cancer cell; genetic regulatory protein; inhibitor/antagonist; mutant; novel; prevent; protein phosphatase inhibitor-2; protein protein interaction; public health relevance; trafficking; tumor

DESCRIPTION (provided by applicant): Activation of NF-?B by anticancer treatments is thought to significantly contribute to tumor survival and clinical resistance. While inhibition of NF-?B is an effective way to sensitize cancer cells to ionizing radiation and chemotherapeutics, the development of clinical NF-?B inhibitors has been difficult. One major barrier in the development of such inhibitors is the pervasive role of NF-?B in mediating a variety of "classic" physiological processes, including innate and adaptive immunity, cell survival, and inflammation. This proposal aims to separate the classical NF-?B pathway from the DNA damage pathway, which contributes to cancer cell survival following treatment with anticancer agents. At the center of this proposal is NEMO, the NF-?B essential modulator. NEMO forms a number of protein-protein interactions required for activation of the DNA damage pathway. While NEMO is also a critical component of the classical pathway, I hypothesize that NEMO protein- protein interactions can be isolated to separate the classical and DNA damage-specific pathways. In order to test this hypothesis, two complementary approaches will be taken. In the biochemical approach, NEMO protein-protein interactions will be carefully mapped to the amino acid level to determine if specific protein- protein interactions can be isolated. In the functional approach, a genetic screen will be used to identify NEMO mutants that show a specific defect in DNA damage-induced NF-?B activation. In addition, novel peptide based inhibitors, developed using preliminary data, will be tested to determine their ability to specifically inhibit the DNA damage pathway. Together, these aims will allow separation of NEMO's role in the activation of the classical and DNA damage-specific pathways. Ultimately, this will facilitate the development of a targeted DNA damage-specific inhibitor of NF-?B activation.

描述（由申请人提供）：抗癌治疗对 NF-κB 的激活被认为对肿瘤存活和临床耐药性有显着贡献。虽然抑制 NF-κB 是使癌细胞对电离辐射和化疗敏感的有效方法，但临床 NF-κB 抑制剂的开发一直很困难。开发此类抑制剂的一个主要障碍是 NF-κB 在介导各种“经典”生理过程中的普遍作用，包括先天性和适应性免疫、细胞存活和炎症。该提案旨在将经典的 NF-κB 通路与 DNA 损伤通路分开，这有助于抗癌药物治疗后癌细胞的存活。该提案的核心是 NEMO，即 NF-κB 重要调节剂。 NEMO 形成许多激活 DNA 损伤途径所需的蛋白质-蛋白质相互作用。虽然 NEMO 也是经典途径的关键组成部分，但我假设可以分离 NEMO 蛋白质-蛋白质相互作用，以区分经典途径和 DNA 损伤特异性途径。为了检验这一假设，将采取两种互补的方法。在生化方法中，NEMO 蛋白质-蛋白质相互作用将被仔细映射到氨基酸水平，以确定是否可以分离特定的蛋白质-蛋白质相互作用。在功能方法中，将使用遗传筛选来识别在 DNA 损伤诱导的 NF-κB 激活中表现出特定缺陷的 NEMO 突变体。此外，将测试利用初步数据开发的新型肽抑制剂，以确定其特异性抑制 DNA 损伤途径的能力。总之，这些目标将允许分离 NEMO 在激活经典途径和 DNA 损伤特异性途径中的作用。最终，这将有助于开发靶向 DNA 损伤特异性 NF-κB 激活抑制剂。