Fluorescent Probes for Studying Copper Oxidation Biology

描述（由申请人提供）：这个项目的长期目标是在分子水平上理解金属离子、氧代谢和氧化应激之间的关系，以及它们对人类健康、衰老和疾病的贡献。这项工作的核心是跟踪活生物系统中氧化还原活性金属离子和氧代谢产物的能力。为了满足这一需求，我们将为体内氧化信号和应激的两个主要贡献者铜和过氧化氢设计荧光传感器，并应用这些新的化学工具来研究铜氧化生物学在神经退行性疾病和年龄相关疾病遗传模型中的作用。铜是调节呼吸、抗氧化剂、结缔组织和神经递质加工的许多氧处理酶所必需的氧化还原活性辅助因子，但其细胞管理不善可通过不受调节的过氧化氢和相关活性氧（ROS）的产生引发氧化应激。随着时间的推移，随后对蛋白质、脂质和核酸的氧化损伤可导致与年龄相关的严重人类疾病（包括癌症、心血管疾病和阿尔茨海默病（AD）等神经退行性疾病）中组织和器官系统的功能下降。然而，H2O2不仅仅是一种混杂的ROS氧化剂，因为越来越多的证据表明，这种氧代谢物的调节产生对于介导控制细胞增殖和/或细胞死亡的事件至关重要。这里设计的化学传感器将提供强大的新化学工具，用于跟踪被怀疑有助于氧化信号和/或应激途径的特定分子物种。具体目标包括：(i)开发选择性和敏感的荧光传感器，用于成像活细胞中不稳定的Cu(l)和Cu(ll), （ii）设计类似的小分子试剂，用于选择性成像H2O2的活细胞，（iii）并应用这些工具研究健康和患病细胞和组织模型中的铜和过氧化物积累，运输和氧化还原功能。研究金属营养素的基础科学及其与生命系统中氧代谢产物的相互作用，有助于了解它们对严重公共卫生疾病的贡献。这些努力对于年龄是危险因素的疾病尤其重要，包括阿尔茨海默氏症和相关的神经退行性疾病，因为它们具有高发生率的金属失调和氧化应激。

DESCRIPTION (provided by applicant): The long-term goal of this program is to understand, at the molecular level, the relationships between metal ions, oxygen metabolism, and oxidative stress, and their contributions to human health, aging, and disease. Central to this effort is the ability to track redox-active metal ions and oxygen metabolites in living biological systems. To meet this need, we will devise fluorescent sensors for copper and hydrogen peroxide, two major contributors to oxidative signaling and stress in the body, and apply these new chemical tools to study the roles of copper oxidation biology in genetic models of neurodegenerative and age-related diseases. Copper is a required redox-active cofactor for many oxygen-processing enzymes that regulate respiration, antioxidants, connective tissue, and neurotransmitter processing, but its cellular mismanagement can trigger oxidative stress through the unregulated production of hydrogen peroxide and related reactive oxygen species (ROS). Over time, subsequent oxidative damage to proteins, lipids, and nucleic acids can lead to the functional decline of tissue and organ systems in serious age-related human diseases, including cancer, cardiovascular disorders, and neurodegenerative diseases such as Alzheimer's disease (AD). H2O2 is more than a promiscuous ROS oxidant, however, as emerging evidence suggests that regulated production of this oxygen metabolite is critical for mediating events that control cell proliferation and/or cell death. The chemosensors devised here will provide powerful new chemical tools for tracking specific molecular species suspected of contributing to oxidative signaling and/or stress pathways. Specific aims include (i) developing selective and sensitive fluorescent sensors for imaging labile Cu(l) and Cu(ll) in living cells, (ii) devising analogous small-molecule reagents for selective live-cell imaging of H2O2, (iii) and applying these tools to study copper and peroxide accumulation, trafficking, and redox function in healthy and diseased cell and tissue models. Studying the basic science of metal nutrients and their interaction with oxygen metabolites in living systems is pertinent to understanding their contributions to serious diseases of public health. These efforts are of particular importance to diseases where age is a risk factor, including Alzheimer's and related neurodegenerative disorders, because they have high incidence of metal misregulation and oxidative stress.