The roles of p53 and MYC dynamics in regulating heterogeneous cell fate responses to genotoxic stress

p53和MYC动力学在调节基因毒性应激的异质细胞命运反应中的作用

• 批准号: 10635353
• 负责人: Eric Batchelor
• 金额: $ 31.83万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10635353
• 关键词: Affect; Apoptosis; Breast Epithelial Cells; Bypass; Cell Cycle Arrest; Cell Fate Control; Cell Fraction; Cells; Cellular Stress; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Damage; Data; Development; Fluorescence Microscopy; Frequencies; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Genes; Genetic Transcription; Genotoxic Stress; Goals; Health; Heterogeneity; Human; Individual; MYC gene; Malignant Neoplasms; Measurement; Mediating; Microscopy; Modeling; Molecular; Mutagens; Mutation; Nucleic Acid Regulatory Sequences; Outcome; Pathologic; Pathway interactions; Pattern; Physiologic pulse; Physiological; Population; Proteins; Proto-Oncogene Proteins c-myc; Proto-Oncogenes; Public Health; Recovery; Regulation; Repression; Research; Resistance; Role; Signal Pathway; Signal Transduction; Stimulus; Stress; System; TP53 gene; Testing; Therapeutic; Time; Tissues; Variant; Work; biological adaptation to stress; cancer cell; cancer therapy; cell killing; cell transformation; chemotherapy; gain of function; gene repression; genome editing; improved; innovation; insight; malignant breast neoplasm; method development; neoplastic cell; novel; novel therapeutic intervention; pharmacologic; prevent; programs; protein expression; response; senescence; temporal measurement; transcription factor; tumor

Project Summary Our long-term goal is to understand how dynamic regulation of signal transduction systems control cellular stress responses. The focus of this proposal is on identifying the mechanisms by which dynamic expression of the transcription factors p53 and MYC coordinately regulate apoptosis and senescence in response to genotoxic stress. Proper regulation of p53 and MYC are of undeniable importance in human health, as their mutation predisposes human cells to cancer. While the regulation and functions of p53 and MYC have been extensively studied, exactly how they generate variable cell fate outcomes in individual cells of a population responding to the same stress remains poorly understood. Our recent studies have shown that the dynamics of p53, the temporal pattern of p53 accumulation and degradation, serves an integral function for controlling MYC levels and cell fate responses to DNA damage. We have shown p53 dynamics to be highly variable between individual cells, but it remains to be determined how such variability contributes to heterogeneous responses to DNA damaging agents, which is critical for understanding tumor cell heterogeneity and evasion of therapies. To answer this question, we will combine time-lapse fluorescence microscopy to quantify p53 and MYC dynamics with quantitative analysis of key transcriptional targets at the single cell level to determine the temporal regulation of the triggering of apoptosis and senescence in response to DNA damage. We will apply this analysis to three conditions: 1.) non-transformed cells expressing normal p53 and MYC, 2.) transformed cells in which MYC expression is elevated over a range of concentrations, and 3.) transformed cells expressing a p53 gain-of-function hotspot mutation. This work will show how p53 and MYC dynamics control initiation of terminal cell fates in physiological and pathological conditions, and it will serve as the basis for approaches to reduce heterogeneous responses to DNA damaging compounds. These results will provide novel insight into the basic functioning of one of the most important stress response pathways in human cells, and are likely to inform innovative therapeutic strategies based on improved timing of the delivery of therapies. More broadly, this study is likely to provide general insights into the growing list of other important signaling pathways that use dynamic regulation.

项目摘要 我们的长期目标是了解信号转导系统的动态调节如何控制蜂窝 压力反应。该提案的重点是确定动态的机制 转录因子的表达p53和MYC协调调节细胞凋亡和衰老 对遗传毒性应激的反应。对人类的适当调节p53和MYC是不可否认的 健康，因为它们的突变使人类细胞患癌症。而p53和 MYC已被广泛研究，正是它们如何在单个中产生可变细胞命运结果 对相同压力的响应的人群的细胞仍然很少理解。我们最近的研究已经 表明p53的动力学是p53积累和降解的时间模式， 控制MYC水平和细胞命运对DNA损伤的反应的积分功能。我们显示了p53 动力学是单个单元之间高度变化的动力学，但仍有待确定这种可变性 有助于对DNA破坏剂的异质反应，这对于理解至关重要 肿瘤细胞异质性和逃避治疗。要回答这个问题，我们将结合延时 荧光显微镜量化p53和MYC动力学，并对钥匙进行定量分析 单个单元格的转录目标，以确定触发的时间调节 响应DNA损伤的凋亡和衰老。我们将将此分析应用于三个条件：1。） 表达正常p53和myc的非转化细胞，2。）转化的myc表达的细胞为 在一定浓度范围内提高，3。）转化表达p53功能的细胞 热点突变。这项工作将显示P53和MYC动力学控制终端细胞命运的启动 在生理和病理条件下，它将作为减少方法的基础 对DNA破坏化合物的异质反应。这些结果将为您提供新颖的见解 人类细胞中最重要的应力反应途径之一的基本功能，很可能 基于改善疗法的时间安排的创新治疗策略。更多的 从广义上讲，这项研究可能会提供对日益增长的其他重要信号清单的一般见解 使用动态调节的途径。