Signaling Mechanisms in Genome Maintenance

基因组维护中的信号机制

• 批准号: 10187261
• 负责人: Marcus Smolka
• 金额: $ 34.28万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10187261
• 关键词: 1-Phosphatidylinositol 3-Kinase; Address; Affect; CRISPR screen; Cell Cycle; Cell Survival; Cells; Clinical Trials; DNA; DNA Damage; DNA Repair; DNA biosynthesis; Defect; Disease; Genetic Diseases; Genetic Materials; Genome; Genome Stability; Genomics; Human Genetics; Immunologic Deficiency Syndromes; Knowledge; Laboratories; Link; Maintenance; Mammals; Mediating; Metabolism; Microscopy; Mutation; Neurologic; Oncogenes; Pharmaceutical Preparations; Phosphotransferases; Predisposition; Proteins; Proteomics; Regulation; Research; Resistance; Resolution; Role; Signal Transduction; Technology; Transact; Work; Yeasts; cancer cell; cancer clinical trial; cancer predisposition; cancer therapy; design; driving force; fitness; genetic approach; genome integrity; genotoxicity; inhibitor/antagonist; novel; phosphoproteomics; programs; replication stress; single molecule; therapeutically effective; tumorigenesis

PROJECT SUMMARY/ABSTRACT The stability of eukaryotic genomes relies on the tight coordination of DNA metabolic processes with DNA repair and the cell cycle. Central to this coordination are elaborate signaling networks mediated by DNA damage signaling kinases. Mutations in these kinases are associated with a range of human genetic disorders linked to cancer predisposition, neurological defects, and immunodeficiency. Selective inhibitors of DNA damage signaling kinases are now being used in over a hundred clinical trials for cancer therapy. However, fundamental questions related to how these kinases maintain genome integrity remain unanswered. The Smolka Laboratory investigates DNA damage signaling, with a major focus on the phosphatidylinositol 3′ kinase (PI3K)‐related kinases (PIKKs) and PIKK‐regulated downstream checkpoint kinases. In addition to pioneering the use of proteomic approaches for the unbiased mapping PIKK substrates, our work uncovered new mechanisms of DNA repair regulation and DNA damage signaling deactivation. Over the next five years, our research program will explore novel non-canonical roles for PIKKs in genome maintenance and DNA replication that have remained obscured for decades. We will address fundamental questions regarding the identity of key effector proteins by which PIKKs control DNA synthesis and DNA repair transactions, establishing the underlying mechanisms of phospho-regulation and implications for manipulating cellular viability, fitness and drug susceptibilities. We will employ extensive phosphoproteomics and genetic approaches, and incorporate single molecule super resolution microscopy, CRISPR screens and genomic technologies. The proposed work will reveal crucial, yet undefined, layers of PIKK-dependent regulation governing genome maintenance and DNA replication in yeast and mammals. Generated knowledge will also have implications for understanding how cells with unstable genomes, such as cancer cells undergoing oncogene-induced replication stress, co-opt PIKK signaling for unrestrained proliferation and resistance to genotoxic insults. Moreover, our findings will illuminate how inhibitors of PIKKs, already in clinical trials, affect cell viability and genomic integrity, thus informing the design of more effective therapeutic strategies.

项目总结/摘要 真核生物基因组的稳定性依赖于DNA代谢过程与DNA的紧密协调 修复和细胞周期。这种协调的核心是由DNA介导的复杂信号网络 损伤信号激酶。这些激酶的突变与一系列人类遗传疾病有关 与癌症易感性、神经缺陷和免疫缺陷有关。选择性DNA抑制剂 损伤信号激酶现在被用于一百多个癌症治疗的临床试验。然而，在这方面， 与这些激酶如何维持基因组完整性有关的基本问题仍然没有答案。的 Smolka实验室研究DNA损伤信号传导，主要关注磷脂酰肌醇3′ 激酶（PI 3 K）相关激酶（PIKK）和PIKK调节的下游检查点激酶。除了 开创性地使用蛋白质组学方法来无偏地绘制PIKK底物，我们的工作揭示了 DNA修复调节和DNA损伤信号失活的新机制。在接下来的五年里， 我们的研究计划将探索PIKK在基因组维护和DNA合成中的新的非经典作用。 几十年来一直被掩盖的复制。我们将讨论有关 PIKK控制DNA合成和DNA修复事务的关键效应蛋白的身份， 建立磷酸调节的潜在机制和操纵细胞的影响， 生存能力、适应性和药物敏感性。我们将采用广泛的磷酸化蛋白质组学和遗传学方法， 方法，并结合单分子超分辨率显微镜，CRISPR屏幕和基因组 技术.这项拟议中的工作将揭示PIKK依赖性调控的关键但未定义的层面 控制酵母和哺乳动物的基因组维持和DNA复制。生成的知识还将 对于理解具有不稳定基因组的细胞，如癌细胞， 癌基因诱导的复制应激，协同PIKK信号传导，用于无限制的增殖和对 遗传毒性的侮辱。此外，我们的发现将阐明已经在临床试验中的PIKK抑制剂， 影响细胞活力和基因组完整性，从而为设计更有效的治疗策略提供信息。