Defining the Multiple Myeloma Kinome

描述(申请人提供)：基因图谱技术使肿瘤细胞的转录组和蛋白质组分析成为可能。这一信息为定义MM细胞增强存活和增殖的分子机制提供了有用的信息。然而，一个同样重要(如果不是更重要)的目标是确定那些参与MM细胞及其支持基质中活跃的信号通路的蛋白质。将酪氨酸、丝氨酸和苏氨酸残基磷酸化的酶在决定多发性骨髓瘤和支持它们的基质细胞的细胞周期进入和存活的信号级联中发挥着重要作用。特别是，了解MM细胞中活跃的信号通路及其支持基质将为了解MM细胞在BM中的生存提供关键信息。我们已经开发并正在应用于纯化细胞的一种新的基于阵列的策略，该策略允许同时检测1152种不同的激酶底物的磷酸化。在这里，我们建议将这项新兴技术应用于分析多发性骨髓瘤及其支持基质中基于磷酸化的细胞信号通路。这项R21/R33阶段性创新应用将分两个阶段进行。在这项应用的R21阶段，目标1和2将验证PepChip技术可以应用于MM细胞及其微环境，以揭示MM中的信号变化。在R33阶段的目标3，我们将使用PepChip技术识别MM患者群体中与临床参数相关的动态组变化，如复发和与预后不良相关的染色体异常。在目标4中，我们将利用支持RAG2XGammaC小鼠原发患者分离株生长的体内模型来确定治疗对MM细胞动态组的影响。本研究将在以下阶段进行。R21/R33分阶段格式：R21阶段：目标1：确定MM细胞和正常浆细胞的动态组。目的2：明确MM与正常骨髓微环境基因组的差异。R33阶段：目标3：确定MM的动态组改变与疾病的临床参数相关。目的4：确定多发性骨髓瘤细胞对治疗的反应中动态组的变化。

DESCRIPTION (provided by applicant): Gene profiling technology has enabled analysis of the transcriptome and proteome of tumor cells. This information has provided useful information with regard to molecular mechanisms that define the enhanced survival and proliferation of MM cells. However, an equally, if not more important, goal is to define those proteins that participate in signaling pathways active in MM cells and their supporting stroma. Enzymes that phosphorylate tyrosine, serins and threonine residues on other proteins play a major role in signaling cascades that determine cell cycle entry and survival in MM and the stromal cells that support them. In particular, knowing the signaling pathways that are active in MM cells and their supporting stroma will provide critical information for understanding MM cell survival in the BM. We have developed and are applying to purified cells a novel array-based strategy that allows the simultaneous detection of phosphorylation for 1152 different kinase substrates. Here we propose to apply this emerging technology to the analysis of phosphorylation-based cell signaling pathways in MM and their supporting stroma. This R21/R33 Phased Innovation application will be pursued in two phases. In the R21 phase of this application Aims 1 and 2 will validate that PepChip technology can be applied to MM cells and their microenvironment to reveal signaling alterations in MM. In Aim 3 of the R33 phase we will use PepChip technology to identify kinome alterations within the MM patient population that are correlated with clinical parameters such as relapse and chromosomal abnormalities associated with poor prognosis. In Aim 4 we will utilize an in vivo model that supports the growth of primary patient isolates in RAG2XGammaC mice to determine the effect of therapeutics on the kinome of MM cells. This study will be pursued in the following.phased R21/R33 format: R21 Phase: Aim 1: Define the kinome of MM cells and normal plasma cells. Aim 2: Define differences in the microenvironmental kinome of MM and normal BM. R33 Phase: Aim 3: Identify kinome alterations in MM correlated with clinical parameters of disease. Aim 4: Identify kinome alterations in MM cells in response to therapeutics in vivo.