Signaling Network Dynamics in Metastatic Prostate Cancer

描述（由申请人提供）：不确定性和争议的特点，诊断和治疗的CaP。迫切需要确定那些威胁生命的CaP形式。这些挑战不能完全面对，直到有一个更好的理解如何在CaP细胞的信号通路不同于他们的正常同行。然而，由于这些生化网络具有非凡的可塑性和鲁棒性，识别这些差异变得复杂。此外，这些并发症在治疗干预方面具有后果。最近几个关键信号蛋白与转移性雄激素非依赖性CaP的相关性为这种疾病的诊断和预后突破提供了希望。然而，缺乏必要的技术工具来直接和同时检测这些酶在其自然环境中的活性。拟议的多学科研究计划旨在开发一种高度集成的分子/分析策略，以评估与CaP相关的信号网络的动态。这些网络的一般反应的差异和相似之处，通过药理学试剂作为细胞类型的函数的途径扰动将被注意到。具体目标包括（1）构建快速启动和评估单细胞中酶活性的半自动化仪器，（2）产生CaP相关信号传导途径节点的选择性报告物，（3）将这些报告物转化为细胞可渗透类似物并将其分子活性与目标1中构建的仪器整合，以及（4）证明这些工程化的报告物可以评估活细胞中的动态信号传导行为。我们还将评估这些报告上游激活剂的敏感性，以及在一系列正常和CaP细胞系中靶向CaP信号网络的抑制剂。公共卫生相关性：目前用于转移性前列腺癌（CaP）的检测、诊断和预后的方法容易被误解，并且不能直接解决导致转化表型的关键生化过程。拟议的多学科合作研究计划将开发新技术，以评估与CaP相关的信号网络的动态。

DESCRIPTION (provided by applicant): Uncertainties and controversies characterize the diagnosis and treatment of CaP. There is a desperate need to identify those forms of CaP that are life threatening. These challenges cannot be fully confronted until there is a better understanding of how the signaling pathways in CaP cells differ from those in their normal counterparts. However, identifying these differences is complicated by the extraordinary plastic and robust nature of these biochemical networks. Furthermore, these complications have consequences in terms of therapeutic intervention. The recent correlation of several key signaling proteins with metastatic androgen-independent CaP offers the promise of diagnostic and prognostic breakthroughs for this disease. However, the requisite technological tools are lacking to directly and simultaneously detect the activity of these enzymes in their natural environment. The proposed multidisciplinary research program seeks to develop a highly integrated molecular/analytical strategy to assess the dynamics of the signaling network associated with CaP. Differences and similarities in the general response of these networks to pathway perturbation by pharmacological agents as a function of cell type will be noted. Specific aims include (1) the construction of a semi-automated instrument that rapidly initiates and assesses enzymatic activity in single cells, (2) the creation of selective reporters of CaP-relevant signaling pathway nodes, (3) the transformation of these reporters into cell permeable analogs and the integration of their molecular activity with the instrumentation constructed in Aim 1, and (4) a demonstration that these engineered reporters can assess dynamic signaling behavior in living cells. We will also evaluate the sensitivity of these reporters to upstream activators, as well as inhibitors that target the CaP signaling network across a series of normal and CaP cell lines. PUBLIC HEALTH RELEVANCE: Current methods for the detection, diagnosis, and prognosis of metastatic prostate cancer (CaP) are subject to misinterpretation and do not directly address key biochemical processes responsible for the transformed phenotype. The proposed multidisciplinary collaborative research program will develop new technology to assess the dynamics of the signaling network linked to CaP.