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

基本信息

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
项目类别：
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

项目摘要

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.

项目概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细胞应激反应。该提案的重点是确定动态的机制转录因子 p53 和 MYC 的表达协调调节细胞凋亡和衰老对遗传毒性应激的反应。 p53 和 MYC 的正确调节对于人类具有不可否认的重要性健康，因为它们的突变会使人类细胞容易患癌症。而p53和p53的调节和功能 MYC 已被广泛研究，确切地说它们如何在个体中产生可变的细胞命运结果群体细胞对相同压力的反应仍知之甚少。我们最近的研究有表明 p53 的动态，即 p53 积累和降解的时间模式，有助于控制 MYC 水平和细胞命运对 DNA 损伤反应的整体功能。我们已经展示了p53 个体细胞之间的动态变化很大，但这种变化是如何产生的仍有待确定有助于对 DNA 损伤剂的异质反应，这对于理解这一点至关重要肿瘤细胞异质性和逃避治疗。为了回答这个问题，我们将结合延时摄影荧光显微镜定量 p53 和 MYC 动态，并对关键蛋白进行定量分析单细胞水平的转录目标以确定触发的时间调节 DNA 损伤引起的细胞凋亡和衰老。我们将此分析应用于三种情况：1.) 表达正常 p53 和 MYC 的非转化细胞，2.) MYC 表达为在一定浓度范围内升高，以及 3.) 表达 p53 功能获得性的转化细胞热点突变。这项工作将展示 p53 和 MYC 动力学如何控制终末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 损伤化合物的异质反应。这些结果将为我们提供新的见解人类细胞中最重要的应激反应途径之一的基本功能，并且可能根据改进的治疗时机提供创新的治疗策略。更多的从广义上讲，这项研究可能会为不断增长的其他重要信号传导提供一般见解使用动态调节的途径。