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-？B的激活被认为显着有助于肿瘤的存活和临床抗性。虽然抑制NF-？B是使癌细胞对电离辐射和化学治疗剂敏感的有效方法，但临床NF-？B抑制剂的发展很困难。这种抑制剂发展的一个主要障碍是NF-？B在介导各种“经典”生理过程中的普遍作用，包括先天和适应性免疫，细胞存活和炎症。该提案旨在将经典的NF-途径与DNA损伤途径分开，该途径在用抗癌药物治疗后有助于癌细胞的存活。该提案的中心是NF-？B基本调制器Nemo。 NEMO形成了激活DNA损伤途径所需的许多蛋白质 - 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蛋白蛋白相互作用以分离经典和DNA损伤特异性途径。为了检验这一假设，将采用两种互补方法。在生化方法中，将仔细映射Nemo蛋白 - 蛋白质相互作用到氨基酸水平，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-？B激活中显示特定缺陷的NEMO突变体。此外，将测试使用初步数据开发的基于肽的新型抑制剂，以确定其专门抑制DNA损伤途径的能力。共同，这些目标将允许Nemo在经典和DNA损伤特异性途径的激活中的作用分离。最终，这将促进NF- b激活的靶向DNA损伤特异性抑制剂的发展。