Dynamics of Signaling Pathways: Mechanism and Function

信号通路的动力学：机制和功能

• 批准号: 9381247
• 负责人: Galit Lahav
• 金额: $ 37.94万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9381247
• 关键词: Affect; Alpha Cell; Behavior; Biological; Cancer Model; Cancer cell line; Cell Cycle; Cell Death; Cell Fate Control; Cell Line; Cell Survival; Cells; Cessation of life; ChIP-seq; Chemotherapy-Oncologic Procedure; DNA Damage; DNA Repair; Data; Development; Drug Exposure; Elements; Event; Evolution; Frequencies; Funding; Gene Activation; Gene Expression; Gene Targeting; Genetic; Genetic Transcription; Genotype; Goals; Growth; Heterogeneity; Human; Human Cell Line; Individual; Knowledge; Lead; Link; Malignant Neoplasms; Maps; Measurement; Measures; Medical; Messenger RNA; Molecular; Monitor; Mus; Mutate; Mutation; Outcome; Pathway interactions; Pattern; Pharmaceutical Preparations; Pharmacotherapy; Phase; Phenotype; Play; Population; Premalignant Cell; Protein p53; Proteins; RNA analysis; Radiation Tolerance; Regulation; Reporter; Role; S Phase; Shapes; Signal Pathway; Signal Transduction; Sister; Site-Directed Mutagenesis; Stress; System; TP53 gene; Testing; Time; Tissues; Tumor Suppressor Proteins; Variant; cancer cell; cellular imaging; chemical genetics; chemotherapy; improved; in vivo; insight; irradiation; killings; mathematical model; mouse model; mutant; novel strategies; predictive modeling; programs; prototype; response; screening; small molecule; targeted cancer therapy; tool; transcriptome sequencing; transcriptomics; tumor

PROJECT SUMMARY Our long-term goal is to understand how the dynamic behavior of biological signals is controlled and how these dynamics affect cellular responses. This proposal focuses on fundamental cellular mechanisms of the p53 signaling network. The p53 system orchestrates cellular responses to environmental insult and spontaneous damage, particularly those that damage DNA. Loss or mutation of the p53 system strongly predisposes human cells to cancer, and is observed in a large fraction of cancers. While the molecular function and regulation of the p53 pathway have been extensively investigated, exactly how wild-type or mutant p53 determines the fate of individual cells, and why cells exposed to the same insult end up having different fates, is poorly understood. Answering these questions requires a quantitative understanding of the order of events in single cells and the causal relationships between cellular background (cancer and non-cancerous cells), p53 status (wild-type or mutated), p53 dynamics and cellular outcomes (growth, arrest or death). In the previous funding period/s we found that the temporal dynamics of p53 (i.e. changes in the levels of p53 over time) play a role in cell fate decisions. Different stresses lead to different p53 dynamics and modulation of p53 dynamics alters cellular outcomes. In addition, cell-to-cell variability in the rate of p53 induction explained factional killing in response to a drug. We now propose to combine quantitative dynamic measurements of p53 and its target genes in single cells, together with single cell RNA-Seq and mathematical modeling to determine the origin of heterogeneity in the p53 response and the transcriptional cascades that link p53 dynamics with specific phenotypic outcomes. We will also investigate the dynamics of p53 across different cancer models, species and tissues in vivo and will search for new tools to perturb p53 dynamics. Our ultimate goal is to understand the regulation and function of p53 under different genetic backgrounds and to identify treatments that will increase the efficacy of irradiation and chemotherapy for cancer cells with specific mutations. Our results will provide new insights into the control and manipulation of the p53 pathway, perhaps the most important pathway protecting human cells against the development of cancer. We anticipate that a detailed quantitative understanding of the p53 circuit, the key circuit controlling the decision to grow or die in single cells, will help us understand why some cells die in response to chemotherapeutic drugs while others survive, and may suggest novel strategies to selectively push cancer cells toward permanent arrest or death. It addition, our study will be help predict the effects of specific drugs on tumors with specific genotypes and will provide a prototype for the analysis, description, and understanding of the dynamics of other signaling pathways in human cells.

项目摘要 我们的长期目标是了解如何控制生物信号的动态行为以及如何控制这些行为 动力学会影响细胞反应。该提案重点是p53的基本细胞机制 信号网络。 p53系统策划了蜂窝对环境侮辱和自发的反应 损坏，尤其是那些损坏DNA的伤害。 p53系统的损失或突变极易于易于体现 人类细胞为癌症，在很大一部分癌症中观察到。而分子功能和 p53途径的调节已得到广泛研究，恰好是野生型或突变体p53 确定单个细胞的命运，以及为什么暴露于同一侮辱的细胞最终会有不同的命运， 理解很差。回答这些问题需要定量了解事件的顺序 单细胞和细胞背景之间的因果关系（癌症和非癌细胞），p53 状态（野生型或突变），p53动力学和细胞结局（生长，逮捕或死亡）。 在上一个资金期间，我们发现p53的时间动力学（即p53的水平变化 随着时间的流逝）在细胞命运决策中发挥作用。不同的应力导致不同的p53动力学和调制 p53动力学改变了细胞结局。此外，解释了p53诱导率的细胞间变异性 派系杀害毒品。现在，我们建议将p53的定量动态测量结合在一起 及其在单细胞中的靶基因以及单细胞RNA-seq和数学建模，以确定 p53响应中异质性的起源和将p53动力学联系起来的转录级联 特定的表型结果。我们还将研究p53在不同癌症模型中的动力学， 体内物种和组织将寻找新工具以扰动p53动力学。我们的最终目标是 了解不同遗传背景下p53的调节和功能，并识别治疗 这将提高辐射和化学疗法对具有特定突变的癌细胞的疗效。 我们的结果将为p53途径的控制和操纵提供新的见解，也许是最多的 保护人类细胞免受癌症发展的重要途径。我们预计这是一个详细的 对p53电路的定量理解，控制决定或死亡的决定 细胞，将有助于我们了解为什么某些细胞在化学治疗药物中死亡而另一些细胞生存，而另一些细胞则会死亡， 并可能提出新的策略，以选择性地将癌细胞推向永久性逮捕或死亡。它 此外，我们的研究将有助于预测特定药物对具有特定基因型的肿瘤的影响，并将 提供一个原型，用于分析，描述和理解其他信号的动态 人类细胞中的途径。