Molecular Imaging Agents for Monitoring Lysine Demethylases in Cell

项目摘要 蛋白质赖氨酸甲基化是由赖氨酸甲基转移酶（KMT或甲基转移酶）控制的可逆过程。 和赖氨酸脱甲基酶（KDM或甲基擦除酶）。大多数关于这些国民党的研究， KDM主要关注它们在细胞核内的功能。这些国民党、国民党， 存在于细胞核中的蛋白质也存在于细胞质中，但对它们的 在这个亚细胞区室中发挥作用。这些酶的异常直接与 癌症、炎症和其他疾病。尽管KDM在不同的亚细胞中至关重要， 在全球分析和有效方法之间存在巨大差距， 实现它。这项研究计划的长期目标是发展一个机械的理解， “赖氨酸甲基化组”维持在亚细胞水平，以及这些化学标记物如何调节细胞 发信号。目前的重点是开发一类新的化学工具，报告KDM活动在生活中 在亚细胞水平上。这个新的化学工具家族将被用来确定KDM的活性 在不同的亚细胞区室，和它们的调节底物，和当地的辅因子。主 生物学的兴趣现在涉及KDM在细胞核外的作用，特别是细胞质， 线粒体，同时在癌症的背景下进行机制研究。拟议的研究包括 四项创新，以亚细胞分辨率解码细胞内KDM活性。第一，发展 新型小分子探针对赖氨酸具有独特的化学选择性， 水解，这是研究细胞中KDM的理想方法。其次是分子显像剂的发展 监测KDM的活性以及它们在细胞内的调节方式。第三，是确定活动的 通过在探针上结合识别元件来识别特定的KDM。四是细胞器的发育 选择性分子成像剂，以研究KDM在不同亚细胞区室中的作用及其 亚细胞定位，这是目前不可能确定与现有的技术。这些探针是 能够确定KDM活性及其在各种疾病状态中的作用， 兴趣本研究将通过提供更好的理解在生物医学领域有广泛的应用 KDM在不同亚细胞区室中发挥作用，疾病的分子机制，因此有助于 发现新的蛋白质生物标志物，以及用于治疗癌症的联合疗法。

PROJECT SUMMARY Protein lysine methylation is a reversible process controlled by lysine methyltransferases (KMTs or methyl writers) and lysine demethylases (KDMs or methyl erasers) in human cells. Most studies on these KMTs and KDMs have focused mainly on their functions inside the nucleus. More than half of these KMTs and KDMs, which are present in the nucleus are also localized in the cytoplasm but very little is known about their functions in this subcellular compartment. The abnormalities in these enzymes are directly associated with cancers, inflammation and other diseases. Despite the critical importance of the KDMs in different subcellular compartments, there is a substantial gap between their global analysis and effective methods available to achieve it. The long-term goal of this research program is to develop a mechanistic understanding of how “lysine methylome” is maintained at a subcellular level and how these chemical markers modulate cell signaling. The current focus is to develop a new class of chemical tools that report on KDM activities in living cells at a subcellular level. This new family of chemical tools will be deployed to determine the activity of KDMs in different subcellular compartments, and their regulation by substrates, and local cofactors. The primary biological interests right now deal with roles of KDMs outside of the nucleus, in particular, the cytoplasm and mitochondria, while pursuing mechanistic studies in the context of cancer. The proposed research contains four innovations to decode the intracellular KDM activities with subcellular resolution. First, is the development of new small molecule probes with unique chemoselectivity towards lysine, and high stability towards hydrolysis, which is ideal for studying KDMs in cells. Second, is the development of molecular imaging agents to monitor the activity of KDMs and how they are regulated inside the cells. Third, is to determine the activity of the specific KDMs by incorporating recognition elements on the probe. Fourth, is the development of organelle selective molecular imaging agents to study the role of KDMs in different subcellular compartments and their subcellular localization, which is currently impossible to determine with existing techniques. These probes are capable of determining KDM activities and their role in various diseased states thus of immense therapeutic interest. This research will have extensive applications in biomedical field by providing a better understanding of KDMs functions in different subcellular compartments, the molecular mechanisms of diseases, thus assist in the discovery of novel protein biomarkers, and combination-therapy for the treatment of cancer.