Nanodiamond Quantum Sensors for Free Radical Detection

用于自由基检测的纳米金刚石量子传感器

• 批准号: 10325762
• 负责人: ALEX I. SMIRNOV
• 金额: $ 25.66万
• 依托单位: ADAMAS NANOTECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-10 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10325762
• 关键词: Affect; Alzheimer' s Disease; Benchmarking; Biological; Biology; Cardiovascular Diseases; Cataract; Cells; Chemicals; Chemistry; Coupled; DNA; Data; Detection; Development; Diabetes Mellitus; Diamond; Disease; Disease Progression; Down Syndrome; Dyes; Electron Spin Resonance Spectroscopy; Environment; Exhibits; Family; Fluorescence; Free Radicals; Gold; Health; Human; Hydroxylamine; Image; In Vitro; Kinetics; Label; Ligands; Lipids; Location; Longitudinal Studies; Magnetism; Malignant Neoplasms; Measurement; Measures; Mediator of activation protein; Methods; Mitochondria; Modeling; Molecular Probes; Nitrogen; Optical Methods; Optics; Outcome; Oxidative Stress; Parkinson Disease; Pathogenesis; Pathway interactions; Phase; Photobleaching; Phototoxicity; Production; Property; Proteins; Protocols documentation; Publications; RNA; Reaction; Reactive Oxygen Species; Reporting; Reproducibility; Research; Research Personnel; Resolution; Role; Salvelinus; Scheme; Signal Pathway; Signal Transduction; Site; Small Business Innovation Research Grant; Source; Specificity; Specimen; Spin Trapping; Superoxides; Surface; System; Tissues; Toxic effect; Xanthine Oxidase; adduct; base; biomaterial compatibility; cellular imaging; chemical stability; chemical synthesis; clinical diagnostics; commercialization; density; design; imaging capabilities; implementation barriers; improved; in vitro testing; macrophage; nanodiamond; nervous system disorder; new technology; novel; oxidative damage; particle; prototype; quantum; sensor; temporal measurement; tool

Summary Reactive oxygen species (ROS) are key mediators in human health but when misregulated can contribute to the progression of many diseases (e.g., cardiovascular disease, Parkinson's disease, Alzheimer's disease, cancer, Down's syndrome, cataract, several neurological disorders, etc.). While biological effects of ROS are thought to be determined by their both spatial (subcellular localization) and temporal (duration of exposure) levels, detailed understanding of site-specific ROS intracellular concentrations and their relationship to the disease pathogenesis is currently missing. The main reason for this is the commercial unavailability of experimental tools to detect and characterize ROS at specific cellular locations with sufficient sensitivity and spatial and temporal resolution. Electron paramagnetic resonance (EPR) is considered to be the gold standard for unambiguous chemical identification of ROS by spin-trapping methods. However, the technical barriers for implementation are high, and efforts toward developing EPR-based imaging of ROS within a biological environment have proven difficult. Methods based on changes in fluorescence emission upon reactions of a dye with ROS are more accessible than EPR. While such optical methods can be readily combined with cellular imaging, the current implementations are riddled with difficulties including lack of specificity in ROS detection, toxicity concerns, artefactual ROS production by the probes themselves, signal variability due to high levels of background fluorescence and, importantly, photobleaching. This phase I proposal advances the field of ROS detection by developing a new family of nanodiamond (ND) based bright fluorescent ROS sensors that will combine the specificity and information content of EPR spin trapping with the advanced imaging capabilities enabled by optical probes without the problems of phototoxicity and photobleaching. Our pathway to commercialization assembles a team with expertise in ND processing and commercialization, development of cutting-edge ROS detection schemes in EPR and chemical synthesis, and expertise in free radical biology and oxidative stress. Phase I is aimed at demonstrating a proof-of-principle prototype ROS sensor which consists of spin-reactive molecules crafted on ND surface and correlating fluorescence and EPR data. The ROS sensor will then be tested in vitro to detect superoxide radical produced by a xanthine oxidase system and then detection and imaging ROS in RAW264.7 macrophages. Benchmarking of ND over conventional ROS optical probes will be aimed to demonstrate advantages of ND ROS sensors in extending the observation period and reducing results' variability. Commercialization of these new ROS detection tools will enable longitudinal studies of site-specific ROS production in cells and tissue to advance the understanding of the roles of ROS and oxidative stress in the pathogenesis and progression of diseases not otherwise achievable. Moreover, the adaption ND-NV-based spin probes to ex vivo clinical diagnostics has high a commercial potential.

概括 活性氧（ROS）是人类健康中的关键介体 许多疾病的进展（例如心血管疾病，帕金森氏病，阿尔茨海默氏病，癌症， Down的综合征，白内障，几种神经系统疾病等）。而ROS的生物学作用被认为是 由它们的空间（亚细胞定位）和时间（暴露时间）水平确定，详细 了解位点特异性ROS细胞内浓度及其与疾病发病机理的关系 目前缺少。造成这种情况的主要原因是实验工具无法检测和 表征具有足够灵敏度以及空间和时间分辨率的特定细胞位置的ROS。 电子顺磁共振（EPR）被认为是明确化学的黄金标准 通过自旋捕获方法识别ROS。但是，实施技术障碍很高，并且 事实证明，在生物环境中开发基于EPR的ROS成像的努力已被证明很困难。 染料与ROS反应后荧光发射变化的方法更容易接近 比EPR。虽然这种光学方法可以很容易地与蜂窝成像相结合，但电流 实施的困难使实施措施包括ROS检测缺乏特异性，毒性问题， 探针本身产生的人工ROS，由于背景高水平而引起的信号变异性 荧光，重要的是光漂白。该阶段我提出了通过 开发一个新的基于纳米原子的家族（ND）的明亮荧光ROS传感器，该传感器将结合起来 EPR旋转诱捕的特异性和信息内容具有由高级成像功能启用 光学探针没有光毒性和光漂白问题。我们的商业途径 组建一个具有ND处理和商业化专业知识的团队，开发尖端ROS EPR和化学合成中的检测方案，以及自由基生物学和氧化应激方面的专业知识。 第一阶段旨在展示原理原型ROS传感器，该原型由自旋反应性组成 在ND表面制成的分子并将荧光和EPR数据相关联。 ROS传感器将是 在体外测试以检测黄嘌呤氧化酶系统产生的超氧化物自由基，然后检测和 在RAW264.7巨噬细胞中成像ROS。常规ROS光学探针的ND基准测试将是 旨在在延长观察期并降低结果中证明ND ROS传感器的优势'' 可变性。这些新的ROS检测工具的商业化将使特定地点的纵向研究 细胞和组织中的ROS产生，以促进对ROS和氧化应激的作用的理解 疾病的发病机理和进展是无法实现的。此外，基于ND-NV的改编自旋 对离体临床诊断的探针具有很高的商业潜力。