Genetic & Metabolic Dissection of the CaMKKbeta Signaling Axis in Prostate Cancer

描述（由申请人提供）：虽然已知异常雄激素受体（AR）信号传导对于前列腺癌的发展是重要的，但也已经变得明显的是，AR信号传导在疾病的致命晚期仍然是活跃的和必要的。尽管已知AR信号在前列腺癌中的重要性，但驱动疾病进展的受体下游过程仍然知之甚少。这种知识差距阻碍了新疗法的开发，特别是对于目前无法治愈的疾病的晚期阶段。因此，长期目标是开发用于治疗前列腺癌的新治疗方法。来自几个独立实验室的先前工作表明，AR信号促进前列腺癌的生长、迁移、侵袭和代谢改变，部分是通过 Ca 2 +/钙调蛋白依赖性蛋白激酶β（CaMKKβ）AMP激活的蛋白激酶（AMPK）信号通路。该提案的主要目标是使用体外和体内模型的组合来定义AR介导的CaMKKβ信号传导在前列腺癌中的特定作用，并测试其是否代表临床前遗传动物模型中的可行药物靶标。中心假设是CaMKKβ轴促进葡萄糖和脂肪酸的病理代谢，因此代表了晚期前列腺癌治疗的新靶点。该假设基于申请人实验室生成的初步和已发布的数据，并得到其他小组研究的大力支持。该假设将通过以下两个具体目标进行验证：目标1：确定AR介导的CaMKKβ-AMPK信号在前列腺癌细胞代谢中的作用。目的2：使用前列腺癌的临床前小鼠模型对CaMKKβ的致病作用进行遗传解剖。在第一个目标下，前列腺癌的分离细胞模型将被用于定义 使用代谢通量分析和综合代谢组学分析技术研究病理代谢中的CaMKKβ、AMPK和候选下游信号传导靶点。在第二个目标中，将使用遗传小鼠模型的组合来描述CaMKKβ在癌症进展的各个阶段中的作用。此外，来自这些研究的肿瘤将进行前述目的中描述的代谢组学分析。这项研究是创新的，因为它测试了AR信号通过糖和脂肪的混杂代谢促进前列腺癌进展的新范式。此外，它使用质谱技术测试这种范例，由于其分辨率提高，将产生肿瘤代谢组的全面检查。这些研究意义重大，因为它们将最终确定CaMKKβ信号传导是否是体内可行的治疗靶点，并确定其活性的潜在代谢生物标志物。最终，预计拟议研究的完成将为后续药物发现工作奠定基础。

DESCRIPTION (provided by applicant): While it is known that aberrant androgen receptor (AR) signaling is important for the development of prostate cancer, it has also become evident that AR signaling remains active and necessary in the deadly advanced stages of the disease. Despite the known importance of AR signaling in prostate cancer, the processes downstream of the receptor that drive disease progression remain poorly understood. This knowledge gap has precluded the development of novel therapies, particularly for the advanced stages of the disease for which there is currently no cure. Thus, the long-term goal is to develop new therapeutic approaches for the treatment of prostate cancer. Previous work from several independent laboratories has suggested AR signaling promotes prostate cancer growth, migration, invasion and altered metabolism in part through a Ca2+/calmodulin-dependent protein kinase kinase beta (CaMKKβAMP-activated protein kinase (AMPK) signaling pathway. The primary goal of this proposal is to use a combination of in vitro and in vivo models to define the specific role(s) of AR-mediated CaMKKβ signaling in prostate cancer and test whether it represents a viable drug target in preclinical genetic animal models. The central hypothesis is that the CaMKKβ axis promotes both glucose and fatty acid pathological metabolism and therefore represents a novel target for advanced prostate cancer therapy. This hypothesis is based on the preliminary and published data generated from the applicant's laboratory and is strongly supported by studies from other groups. The hypothesis will be tested with the following two specific aims: Aim 1: Determine the role of ARmediated CaMKKβ-AMPK signaling in prostate cancer cellular metabolism. Aim 2: Genetic dissection of the pathogenic role of CaMKKβ using preclinical mouse models of prostate cancer. Under the first aim, isolated cellular models of prostate cancer will be used to define the specific roles of CaMKKβ, AMPK and candidate downstream signaling targets in pathological metabolism using metabolic flux analysis and comprehensive metabolomic profiling techniques. In the second aim, a combination of genetic mouse models will be used to delineate the role of CaMKKβ in various stages of cancer progression. Further, tumors derived from these studies will be subjected to the metabolomic profiling described in the previous aim. The research is innovative because it tests the novel paradigm that AR signaling promotes prostate cancer progression through the promiscuous metabolism of both sugars and fats. Further, it tests this paradigm using mass spectroscopic techniques that, due to their enhanced resolution, will yield a comprehensive examination of the tumor metabolome. These studies are significant because they will conclusively determine whether CaMKKβ signaling is a viable therapeutic target in vivo and also identify potential metabolic biomarkers of its activity. Ultimately, it is anticipated that the completion of the proposed studis will set the foundation needed for subsequent drug discovery efforts.