The role of dynamics in defining the limits of normal developmental signaling.

动力学在定义正常发育信号限制中的作用。

• 批准号: 9324287
• 负责人: John G. Albeck
• 金额: $ 31.48万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9324287
• 关键词: Affect; Automobile Driving; Behavior; Cancer Biology; Cell Line; Cell Proliferation; Cell model; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; Complex; Complication; Computer Simulation; Data; Defect; Development; Disease; Dose; Drug Targeting; Drug usage; Engineering; Event; Fos-Related Antigens; Frequencies; Gene Expression; Gene Expression Process; Gene Expression Profile; Gene Targeting; Genes; Genetic Transcription; Germ-Line Mutation; Heart Abnormalities; Homeostasis; Human; Human Development; Human Genome; Hyperactive behavior; Imaging Techniques; Impaired cognition; In Vitro; Individual; Inherited; Kinetics; Knock-in; Lead; Link; Live Birth; Malignant Neoplasms; Measurement; Measures; Methods; Modeling; Molecular; Monitor; Mutation; Nature; Outcome; Pathologic; Pathology; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Pharmacology; Phenotype; Phosphotransferases; Play; Probability; Process; Proteins; Regulation; Reporter; Role; Series; Signal Transduction; Surveys; Syndrome; System; Testing; Therapeutic; Time; Tissues; Translations; Variant; Work; cancer genetics; cancer risk; cell behavior; cell motility; cognitive development; developmental disease; disease-causing mutation; genetic regulatory protein; human disease; imaging platform; inhibitor/antagonist; live cell imaging; mRNA Differential Displays; mathematical model; migration; mutant; preference; programs; public health relevance; repaired; response; single cell analysis; transmission process

 DESCRIPTION (provided by applicant): Signaling by the Ras/ERK pathway controls cell proliferation, migration, and differentiation. Proper function of this pathway is essential for human development and homeostasis. Sporadic mutations in the pathway play a central role in many cancers, while hereditary mutations cause a series of congenital syndromes, termed RASopathies, which result in impaired cognitive development, cardiac malformations, and increased risk of cancer. A major unanswered question is what differentiates normal from pathological Ras/ERK signaling. It is known that the dynamic pattern of ERK activity - including the strength, frequency, and duration of its activity - are essential to proper signaling. However, the standard methods for measuring ERK activity lack the single-cell precision needed to resolve these essential details. In this project, we will use live-cell imaging, which allows nearl continuous monitoring of thousands of cells simultaneously, to collect data on mutant-driven ERK signaling that is far more accurate and detailed than was previously available. We will focus on the mutations found in the RASopathies, where their role as a single gene driving pathological effects in development is more clearly defined than in cancer, where mutations in many other genes are a complication. We will use our imaging platform to compare for the first time the changes in ERK signaling resulting from disease-causing mutations at the single cell level. Using multiple in vitro systems to replicate cellular processes involved in development, we will determine how these changes modify cell proliferation, migration, and differentiation. We will then dissect the mechanisms underlying these phenotypic changes at the level of gene expression, using a new class of reporters that are integrated directly into the genomes of human cells. At the levels of kinase kinetics, gene expression, and cell behavior, we will quantify how mutant cells respond to multiple Ras pathway inhibitors, which are now being considered as treatments for the RASopathies. This work will have several important outcomes. First, it will reveal the quantitative boundaries of signal behavior that are compatible with normal function, allowing us to understand how Ras pathway mutations lead to disease, and why some mutations are more severe than others. Secondly, it will allow us to make rational choices about which drugs to give to patients with different mutations, so that treatment can be personalized to best normalize each individual's specific signaling patterns. Finally, it will result in a mathematical model of the link between kinase activity and downstream gene expression programs that will allow us to better understand developmental programs and engineer desired cellular responses using existing drugs that target kinase activity.

 描述（由申请人提供）：Ras/ERK 通路的信号传导控制细胞增殖、迁移和分化，该通路的正常功能对于人类发育和稳态至关重要，而该通路中的零星突变在许多癌症中发挥着核心作用。遗传性突变会导致一系列先天性综合征，称为 RASopathies，导致认知发育受损、心脏畸形和癌症风险增加，一个尚未解答的主要问题是差异是什么。病理性 Ras/ERK 信号传导正常 众所周知，ERK 活动的动态模式（包括其活动的强度、频率和持续时间）对于正确的信号传导至关重要。 测量 ERK 活性的标准方法缺乏解决这些基本细节所需的单细胞精度。在这个项目中，我们将使用活细胞成像来收集突变驱动的数据，该成像可以几乎连续地同时监测数千个细胞。我们将重点关注在 RASopathies 中发现的 ERK 信号传导，它们作为驱动发育中病理效应的单个基因的作用比在癌症中更明确，而在癌症中，许多其他疾病都有突变。基因是一个并发症。我们将使用我们的成像平台首次在单细胞水平上比较由致病突变引起的 ERK 信号变化，使用多个体外系统复制参与发育的细胞过程，我们将确定这些变化如何改变细胞。我们会增殖、迁移和分化。 然后，使用直接整合到人类细胞基因组中的一类新产物，在基因表达水平上剖析这些表型变化背后的机制。在激酶动力学、基因表达和细胞行为水平上，我们将进行量化。 突变细胞如何对多种 Ras 通路抑制剂做出反应，目前正在考虑将其作为 RASopathies 的治疗方法。首先，它将揭示与正常功能兼容的信号行为的定量界限。了解 Ras 通路突变如何导致疾病，以及为什么某些突变比其他突变更严重，这将使我们能够合理选择向具有不同突变的患者提供哪些药物，以便可以进行个性化治疗，以最好地使每种突变正常化。最后，个人的特定信号模式。它将产生激酶活性和下游基因表达程序之间联系的数学模型，使我们能够更好地理解发育程序并利用现有的针对激酶活性的药物来设计所需的细胞反应。