Signaling Mechanisms in Genome Maintenance

基因组维护中的信号机制

基本信息

批准号：
10374160
负责人：
Marcus Smolka
金额：
$ 59.21万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
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/10374160
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 novel phosphoproteomics programs replication stress single molecule therapeutically effective tumorigenesis

项目摘要

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.

项目概要/摘要真核基因组的稳定性依赖于DNA代谢过程与DNA的紧密协调这种协调的核心是由 DNA 介导的复杂的信号网络。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 DNA 免疫缺陷有关。目前，用于癌症治疗的一百多个临床试验中使用的信号激酶已被破坏。与这些激酶如何维持基因组完整性相关的基本问题仍未得到解答。 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 (PI3K) 相关激酶 (PIKK) 和 PIKK 调节的下游检查点激酶。我们的工作发现，开创性地使用蛋白质组学方法来绘制 PIKK 底物的无偏差图谱 DNA 修复调节和 DNA 损伤信号失活的新机制。我们的研究计划将探索 PIKK 在基因组维护和 DNA 中的新的非规范作用我们将解决几十年来一直模糊的复制问题。 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以及对细胞操纵的影响我们将采用广泛的磷酸蛋白质组学和遗传学。方法，并结合单分子超分辨率显微镜、CRISPR 筛选和基因组拟议的工作将揭示 PIKK 依赖的关键但未定义的监管层。控制酵母和哺乳动物的基因组维护和 DNA 复制也将产生知识。对于理解基因组不稳定的细胞（例如癌细胞）如何经历变化具有重要意义癌基因诱导的复制应激，选择 PIKK 信号传导以实现无限制的增殖和抗性此外，我们的研究结果将阐明已经在临床试验中的 PIKK 抑制剂如何影响基因毒性。影响细胞活力和基因组完整性，从而为更有效的治疗策略的设计提供信息。