Fluorescent Probes for Studying Copper Oxidation Biology

描述(申请人提供)：该项目的长期目标是在分子水平上了解金属离子、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。这项工作的核心是能够跟踪生物系统中具有氧化还原活性的金属离子和氧代谢物。为了满足这一需求，我们将设计铜和过氧化氢的荧光传感器，这是体内氧化信号和压力的两个主要贡献者，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。铜是许多氧处理酶所必需的氧化还原活性辅因子，这些酶调节呼吸、抗氧化剂、结缔组织和神经递质的处理，但它的细胞管理不善可以通过不受调控的过氧化氢和相关的活性氧物种(ROS)的产生来触发氧化应激。随着时间的推移，蛋白质、脂质和核酸随后的氧化损伤可能会导致与年龄相关的严重人类疾病的组织和器官系统的功能下降，包括癌症、心血管疾病和神经退行性疾病，如阿尔茨海默病(AD)。然而，过氧化氢不仅仅是一种混杂的ROS氧化剂，因为新出现的证据表明，这种氧代谢产物的调节产生对于调节控制细胞增殖和/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来追踪被怀疑参与氧化信号和/或应激途径的特定分子物种。具体目标包括：(I)开发选择性和灵敏的荧光传感器，用于活体细胞中不稳定的铜(L)和铜(111)的成像；(Ii)设计类似的小分子试剂，用于选择性地对过氧化氢进行活细胞成像；(Iii)应用这些工具来研究铜和过氧化氢在健康和疾病细胞和组织模型中的积累、运输和氧化还原功能。研究生命系统中金属营养素及其与氧代谢产物的相互作用的基础科学，对于了解它们对严重的公共健康疾病的贡献具有重要意义。这些努力对年龄是风险因素的疾病特别重要，包括阿尔茨海默氏症和相关的神经退行性疾病，因为它们有很高的金属调节不当和氧化应激的发生率。

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.