Dynamics and Topology of Phosphotyrosine-SH2 Interaction Networks

描述（由申请人提供）：与酪氨酸激酶连接的细胞表面受体控制许多重要的细胞活动，包括增殖、分化和运动。酪氨酸激酶信号失调是许多人类癌症的共同特征，因此酪氨酸激酶及其下游效应物是开发治疗癌症新药的靶点。然而，为了充分利用这些有希望的新疗法，我们需要了解酪氨酸激酶信号网络如何在系统水平上处理信息。虽然在开发描述酪氨酸激酶信号网络的定量模型方面取得了相当大的进展，但由于缺乏关于酪氨酸磷酸化变化如何与含有模块化磷酸酪氨酸结合域的下游效应物耦合的定量信息，这些努力受到严重阻碍。这个合作项目的目标是利用新的实验方法来直接解决这一知识差距。具体来说，我们将使用SH2分析，这是一种与基于质谱的方法高度互补的磷蛋白质组学方法，来量化受体酪氨酸激酶激活时特定效应蛋白结合位点的动态变化。将比较对不同受体和不同细胞类型的反应，从而使系统级行为与生物输出相关联。我们还将使用单分子成像方法来监测细胞内环境中特定效应器与受体的耦合。这些研究将为受体信号复合物的相互作用动力学提供前所未有的见解，这将使酪氨酸激酶信号的更强大和准确的模型成为可能。相关性（见说明）：具有酪氨酸激酶活性的受体发出的信号在许多人类疾病，特别是癌症中起着重要作用。精确描述这些受体使用的信号机制的定量计算机模型将在设计新的癌症治疗方法和决定哪些患者将从这些治疗（个体化医学）中受益最大方面非常有用。提出的研究使用创新的实验方法来揭示建立更强大和准确的模型所必需的新的机制见解。

DESCRIPTION (provided by applicant): Cell surface receptors linked to tyrosine kinases control a host of important cellular activities, including proliferation, differentiation, and motility. Disregulated tyrosine kinase signaling is a common feature of many human cancers, thus tyrosine kinases and their downstream effectors are targets for the development of new drugs for the treatment of cancer. In order to take full advantage of such promising new therapies, however, we need an understanding of how tyrosine kinase signaling networks process information on a systems level. While considerable progress has been made in developing quantitative models describing tyrosine kinase signaling networks, these efforts are severely hampered by a lack of quantitative information on how changes in tyrosine phosphorylation are coupled to their downstream effectors containing modular phosphotyrosine binding domains. The goals of this collaborative project are to take advantage of new experimental approaches to address this gap in knowledge directly. Specifically, we will use SH2 profiling, a phosphoproteomic approach that is highly complementary to mass spectrometry-based methods, to quantify dynamic changes in binding sites for specific effector proteins upon receptor tyrosine kinase activation. Responses to different receptors and in different cell types will be compared, allowing systems-level behavior to be correlated with biological outputs. We will also use single-molecule imaging methods to monitor the coupling of specific effectors to receptors in the intracellular environment. These studies will afford unprecedented insight into the interaction dynamics of receptor signaling complexes that will enable much more powerful and accurate models of tyrosine kinase signaling. RELEVANCE (See instructions): Signaling from receptors with tyrosine kinase activity plays an important role in a number of human diseases, in particular cancer. Quantitative computer-based models that accurately describe the signaling mechanism used by these receptors will be very useful in designing new therapies for cancer and in deciding which patients will benefit most from those therapies (individualized medicine). The proposed studies use innovative experimental approaches to reveal new mechanistic insights necessary for building more powerful and accurate models.