Chemical Probes for Imaging Reactive Sulfur, Oxygen, and Nitrogen Species in Living Cells and Clinical Samples

 描述(由申请人提供)：活性硫、氧和氮(RSON)是内源性小分子，在细胞信号中发挥基本作用，在从癌症到神经退化到糖尿病的各种疾病中调节不当。这个超家族的信号分子包括一氧化氮、硫化氢、过氧化氢和许多其他物质。尽管参与了几乎所有的生理过程，但我们对它们复杂和相互交织的作用的了解仍处于起步阶段，这在很大程度上是由于缺乏方法。 监测活细胞和人类临床样本中的这些暂时性物种。该项目旨在使用高度创新的化学方法开发响应荧光染料，用于精确实时跟踪特定的RSON物种，包括硫化氢、过氧亚硝酸盐和氮氧基，并使用这些探针在肺癌细胞模型和人类唾液/呼出的呼气冷凝物中研究它们的产生。具体地说，我们的目标是：1.开发基于反应的探针来检测和成像细胞和临床样本中的RSON种类。对于某些RSON物种，特别是硫化氢、过氧亚硝酸根和氮氧基的检测，缺乏生物相容的方法。利用我们在合成有机化学方面的专业知识，我们将通过发明新的基于反应的探测器来利用荧光、化学发光和核磁共振技术来检测和成像这些物种，从而弥合这一差距。2.研究RSON在疾病细胞模型中的作用。尽管在癌症模型中已经对普遍存在的信号分子一氧化氮进行了很好的研究，但其他RSON物种的产生和作用仍然不完全清楚。我们将使用新开发的和最先进的基于反应的探针来表征呼吸道炎症的细胞模型中RSON物种的复杂细胞化学。3.开发和验证用于检测和管理疾病的护理点诊断方法。RSON化学有可能成为一种强大的诊断和预后标志物。事实上，呼出的一氧化氮和过氧化氢是监测哮喘和其他呼吸道疾病的公认生物标记物，但家庭和护理点监测仍然是一个重大障碍。我们将开发基于智能手机的创新护理点RSON检测技术，以监测唾液和呼出的呼吸冷凝物中这些物种的水平。

 DESCRIPTION (provided by applicant): Reactive sulfur, oxygen, and nitrogen (RSON) species are endogenous small molecules that play fundamental roles in cellular signaling and are misregulated in diseases ranging from cancer to neurodegeneration to diabetes. This super-family of signaling molecules includes nitric oxide, hydrogen sulfide, hydrogen peroxide, and many others. Despite being involved in nearly all physiological processes, our understanding of their complex and intertwined roles remains in its infancy due, in large part, to a lack of methods to monitor these transient species in living cells and human clinical samples. This project aims to use highly innovative chemistry to develop responsive fluorescent dyes for the precise real-time tracking of specific RSON species, including hydrogen sulfide, peroxynitrite, and nitroxyl, and to use these probes to investigate their production in cellular models of lung cancer and in human saliva/exhaled breath condensates. Specifically, we aim to: 1. Develop reaction-based probes to detect and image RSON species in cells and clinical samples. There is a lack of biologically compatible methods for the detection of certain RSON species, particularly hydrogen sulfide, peroxynitrite, and nitroxyl. Leveraging our expertise in synthetic organic chemistry, we will bridge this gap by inventing new reaction-based probes to detect and image these species using fluorescence, chemiluminescence, and nuclear magnetic resonance techniques. 2. Investigate the role of RSON species in cellular models of disease. Although the ubiquitous signaling molecule nitric oxide has been well studied in cancer models, the production and roles of other RSON species remain incompletely understood. We will use newly developed and state-of-the-art reaction-based probes to characterize the complex cellular chemistry of RSON species in a cellular model of airway inflammation. 3. Develop and validate point-of-care diagnostics for the detection and management of disease. RSON chemistry has the potential to be a powerful diagnostic and prognostic marker. Indeed, exhaled nitric oxide and H2O2 are established biomarker for monitoring asthma and other respiratory ailments, but home and point-of-care monitoring remains a significant obstacle. We will develop innovative point-of-care smartphone-based RSON detection techniques for monitoring the levels of these species in the saliva and exhaled breath condensates.