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.

总结 活性氧（ROS）是人类健康的关键介质，但当调节不当时，可能会导致 许多疾病的进展（例如，心血管疾病，帕金森病，阿尔茨海默病，癌症， 唐氏综合征、白内障、几种神经系统疾病等）。虽然ROS的生物效应被认为是 由其空间（亚细胞定位）和时间（暴露持续时间）水平决定，详细 了解特定位点的ROS细胞内浓度及其与疾病发病机制的关系 现在失踪了其主要原因是商业上无法获得实验工具来检测和 以足够的灵敏度以及空间和时间分辨率表征特定细胞位置处的ROS。 电子顺磁共振（EPR）被认为是明确化学物质的金标准。 通过自旋捕获方法鉴定ROS。然而，实施的技术障碍很高， 在生物环境中开发基于EPR的ROS成像的努力已被证明是困难的。 基于染料与ROS反应后荧光发射变化的方法更容易获得 比EPR。虽然这样的光学方法可以容易地与细胞成像组合，但是目前的光学成像方法不适合于细胞成像。 实施过程充满了困难，包括ROS检测缺乏特异性，毒性问题， 探针本身产生的人为ROS，由于高水平的背景导致的信号变异性 荧光和重要的光漂白。该第一阶段提案通过以下方式推进了ROS检测领域： 开发一种新的基于纳米金刚石（ND）的明亮荧光ROS传感器家族，该传感器将联合收割机 EPR自旋捕获的特异性和信息内容，具有先进的成像能力， 没有光毒性和光漂白问题的光学探针。我们的商业化之路 组建了一支在ND加工和商业化方面具有专业知识的团队，开发尖端的ROS EPR和化学合成的检测方案，以及自由基生物学和氧化应激方面的专业知识。 第一阶段的目的是展示一个原理验证原型ROS传感器，它由自旋反应 分子制作的ND表面和相关的荧光和EPR数据。然后ROS传感器将 体外试验检测黄嘌呤氧化酶系统产生的超氧自由基， RAW264.7巨噬细胞中的ROS成像。将ND与传统ROS光学探头进行基准测试， 目的是证明ND ROS传感器在延长观察周期和减少结果方面的优势， 可变性这些新的ROS检测工具的商业化将使特定位点的纵向研究成为可能。 细胞和组织中ROS的产生，以促进对ROS和氧化应激在细胞和组织中作用的理解。 发病机制和疾病的进展，否则无法实现。此外，基于ND-NV的自适应自旋 用于离体临床诊断的探针具有很高的商业潜力。