Defining the Multiple Myeloma Kinome

描述（由申请人提供）：基因谱分析技术已经能够分析肿瘤细胞的转录组和蛋白质组。该信息提供了关于定义MM细胞增强的存活和增殖的分子机制的有用信息。然而，一个同样的，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性信号通路的蛋白质。使其他蛋白质上的酪氨酸、丝氨酸和苏氨酸残基磷酸化的酶在决定MM和支持它们的基质细胞中的细胞周期进入和存活的信号级联中起主要作用。特别是，了解MM细胞及其支持基质中活跃的信号通路将为了解BM中MM细胞的存活提供关键信息。我们已经开发并应用于纯化的细胞一种新的阵列为基础的策略，允许同时检测1152个不同的激酶底物的磷酸化。在这里，我们建议将这一新兴技术应用于MM及其支持基质中基于磷酸化的细胞信号通路的分析。R21/R33分阶段创新应用将分两个阶段进行。在本申请的R21阶段，目标1和2将验证PepChip技术可应用于MM细胞及其微环境以揭示MM中的信号传导改变。在R33阶段的目标3中，我们将使用PepChip技术来鉴定MM患者群体中与临床参数相关的激酶组改变，所述临床参数例如与不良预后相关的复发和染色体异常。在目标4中，我们将利用支持RAG 2XGammaC小鼠中原代患者分离株生长的体内模型来确定治疗剂对MM细胞激酶组的影响。本研究将按照以下阶段R21/R33格式进行：R21阶段：目标1：定义MM细胞和正常浆细胞的激酶组。目的2：确定MM和正常BM的微环境激酶组的差异。R33阶段：目的3：确定MM中与疾病临床参数相关的激酶组改变。目的4：鉴定MM细胞中响应于体内治疗的激酶组改变。

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.