Core B Proteomics & Biostatistics

核心 B 蛋白质组学

• 批准号: 9074406
• 负责人: Lee M Graves
• 金额: $ 34.98万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-22 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9074406
• 关键词: Adhesions; Alleles; Automobile Driving; Autophagocytosis; Biometry; Cancer Etiology; Cell Proliferation; Cell Surface Receptors; Cells; Cessation of life; Collaborations; Complement; Complex; Data; Development; Ectopic Expression; Evaluation; Event; FRAP1 gene; Family; Feedback; Frequencies; Gene Family; Goals; Growth; HRAS gene; Human; In Vitro; Inflammation; KRAS2 gene; Knowledge; Lead; Link; MEKs; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mitogen-Activated Protein Kinases; Mutate; Mutation; Oncogenes; Output; Pathway interactions; Pharmaceutical Preparations; Phase; Phosphorylation; Protein Array; Protein Array Analysis; Protein Isoforms; Protein Kinase; Protein Tyrosine Kinase; Proteins; Proteomics; Proto-Oncogene Proteins c-akt; RNA Interference; Receptor Protein-Tyrosine Kinases; Role; Signal Transduction; Signaling Protein; Up-Regulation; autocrine; base; cancer cell; cancer initiation; cancer therapy; feeding; in vivo; inhibitor/antagonist; innovation; innovative technologies; melanoma; migration; mutant; novel; prevent; programs; protein activation; tumorigenesis

DESCRIPTION The aim of Core B is to provide innovative technologies to profile RAS mutation-specific effector signaling. RAS effector signaling is complex and involves RAS interaction with a multitude (>10) of functionally diverse downstream effectors. While our current understanding of RAS effector utilization is advanced, it is also far from complete. To date, four effector families have been implicated in driving RAS-dependent cancer initiation and growth. Each effector network includes protein kinases. There is also significant crosstalk between the effector networks. Furthermore, these networks are highly dynamic, with complex feed-forward and feedback mechanisms. Classically, RAS effector signaling is profiled by evaluation of the two canonical effector pathways, the RAF-MEK-ERK mitogen-activated protein kinase cascade and the PI3K-AKT-mTOR prosurvival signaling network, using the phosphorylated state of ERK and AKT as readouts. However, it is now clear that these analyses alone fail to provide an adequate determination of RAS effector signaling. Since a major goal of this Program Project is the determination of RAS mutant-specific effector signaling, unbiased kinome-wide analyses are needed to accomplish this goal. Core B provides two innovative proteomics-based experimental platforms to accomplish this. First, Multiplexed Inhibitor Beads (MIBs) and Mass Spectroscopy (MIB/MS) analyses provide kinome-wide profiling of dynamic changes in protein kinase activity. Our preliminary studies applying MIB/MS to characterize such changes upon KRAS suppression identified protein kinases not previously known as components of RAS effector signaling, demonstrating the potential for this platform to identify novel RAS effector signaling outputs. Second, Reverse Phase Protein Array (RPPA) analyses will profile RAS-dependent changes in protein phosphorylation and activation states in cancer cell signaling networks. Additionally, a recently developed innovative advance in RPPA enabling the profiling of the activation state of interacting proteins will also be applied. The types of data generated using each experimental platform are highly complementary. We expect that together they will define novel RAS mutation- specific effector signaling networks.

描述 核心B的目标是提供创新技术来分析RAS突变特异性效应信号。 RAS效应器信号是复杂的，涉及RAS与多种(&gt；10)不同功能的RAS相互作用 下游效应器。虽然我们目前对RAS效应器利用的理解是先进的，但它也远远 从完整开始。到目前为止，已有四个效应家族参与了RAS依赖性肿瘤的发生 和增长。每个效应器网络都包括蛋白激酶。还存在显著的串扰 效应器网络。此外，这些网络是高度动态的，具有复杂的前馈和反馈 机制。传统上，RAS效应器信号是通过评估两个典型的效应器来描述的 RAF-MEK-ERK丝裂原活化蛋白激酶级联通路与PI3K-AKT-mTOR的生存 信号网络，使用ERK和AKT的磷酸化状态作为读数。然而，现在很明显， 这些分析本身并不能提供RAS效应信号的充分确定。因为一个主要目标是 该计划的项目是确定RAS突变体特有的效应信号，在全基因组范围内无偏倚 为了实现这一目标，需要进行分析。Core B提供了两个创新的基于蛋白质组学的实验 实现这一点的平台。第一，复合抑制剂微珠(MIB)和质谱学(MIB/MS) 分析提供了蛋白激酶活性动态变化的动态组范围的轮廓。我们的初步研究 应用MIB/MS表征KRAS抑制后的这种变化 以前被称为RAS效应器信号的组件，展示了该平台的潜力 识别新的RAS效应器信号输出。第二，反相蛋白质阵列(RPPA)分析将 癌细胞信号转导中蛋白磷酸化和激活状态依赖于蛋白Ras的变化 网络。此外，最近在RPPA方面开发的一项创新进展使能够对 相互作用蛋白质的激活状态也将被应用。使用每个对象生成的数据类型 实验平台互补性强。我们预计他们将共同定义新的RAS突变- 特定的效应器信令网络。