Tools for the Detection of Ethylene

乙烯检测工具

• 批准号: 10018050
• 负责人: Brian Michel
• 金额: $ 15.07万
• 依托单位: UNIVERSITY OF DENVER (COLORADO SEMINARY)
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10018050
• 关键词: Alkenes; Biological; Biological Assay; Biological Markers; Cardiovascular Diseases; Cell Culture Techniques; Cells; Cellular Assay; Communities; Complex; Custom; Detection; Development; Disease; Environment; Ethylenes; Exhalation; Fluorescent Probes; Gases; Goals; Human; Hydrophobicity; Hydroxyl Radical; Investigation; Kinetics; Lasers; Ligands; Lipid Peroxidation; Lipid Peroxides; Location; Malignant Neoplasms; Mammalian Cell; Mammals; Measures; Membrane; Methods; Microscopic; Mission; Modification; National Institute of General Medical Sciences; Nature; Neurodegenerative Disorders; Oxidative Stress; Pharmaceutical Chemistry; Plant Growth Regulators; Play; Production; Property; Public Health; Reactive Oxygen Species; Reporting; Research; Research Proposals; Resolution; Role; Ruthenium; Sampling; Signal Transduction; Site-Directed Mutagenesis; Stress; Structure; Structure-Activity Relationship; System; Technology; Transition Elements; Translating; Variant; base; catalyst; chemical synthesis; design; experimental study; fluorophore; improved; instrument; lipophilicity; metal complex; pi bond; small molecule; subcellular targeting; tool

7. Project Summary/Abstract While ethylene has long been known as an important plant hormone it has also been demonstrated to be produced in mammals as a result of oxidative stress that is hallmark to numerous diseases. In particular ethylene arises from the radical fragmentation of lipid peroxides and/or intermediates in their formation. The formation of lipid peroxides is a result of reactive oxygen species, which are implicated as playing stress or signaling roles in numerous diseases including cancer, cardiovascular disease, and neurodegenerative diseases amongst others. While there are some sophisticated spectroscopic methods for sensitively measuring the biomarker ethylene in exhaled breath, these approaches are necessarily limited in spatial resolution and complexity of sample. Recently, our group has developed a profluorescent chemodosimeter that is capable of detecting ethylene in live cells; however the current sensitivity is insufficient for detection of endogenous ethylene. Therefore the overall goal of this exploratory research proposal is to determine if structural and targeting modifications can provide the necessary sensitivity to study the endogenous production of ethylene and any potential signaling roles. To achieve this goal we aim to (1) structurally optimize the ligands about the ruthenium and tune the photophysical properties of the appended fluorophore; and (2) to use subcellular targeting of the probe to cellular domains where ethylene is expected to be present in higher concentrations. It is expected that the proposed research will result in probes with significantly improved limit of detection (~2 orders of magnitude) in the range that might be expected for endogenous ethylene levels in disease states. This will primarily be accomplished through synthesis of modified probes, characterization of resulting properties, and localization studies in live cells. This exploratory research is expected to evaluate the feasibility of detecting endogenous ethylene levels and lay the groundwork for further investigations into ethylene's production at the cellular level. The ability to detect ethylene with spatial resolution not available using current approaches would provide a broadly applicable tool capable of reporting lipid peroxidation. 1

7.项目摘要/摘要 虽然乙烯长期以来一直被认为是一种重要的植物激素，但它也被证明 在哺乳动物中由于氧化应激而产生，氧化应激是多种疾病的标志。在……里面 特定的乙烯产生于过氧化脂质和/或其中间产物的自由基碎裂。 队形。过氧化脂质的形成是活性氧物种的结果，这与 在许多疾病中扮演压力或信号角色，包括癌症、心血管疾病和 神经退行性疾病等。虽然有一些复杂的光谱方法 为了灵敏地测量呼气中的生物标记物乙烯，这些方法是必要的 在空间分辨率和样本复杂性方面受到限制。最近，我们小组开发了一种 前荧光化学剂量计，能够检测活细胞中的乙烯；但电流 对于内源乙烯的检测，灵敏度不够。 因此，这一探索性研究提案的总体目标是确定结构和 靶向修饰可以提供必要的敏感性来研究内源产物 乙烯和任何潜在的信号作用。为了实现这一目标，我们的目标是(1)从结构上优化 有关Ru的配体，并调节附加的荧光团的光物理性质；以及(2) 使用亚细胞靶向将探针定位到预期存在乙烯的细胞域 浓度更高。 预计拟议的研究将导致探测器的限制大大改善 在可能预期的内源乙烯水平范围内检测(~2个数量级) 疾病状态。这将主要通过合成修饰的探针来实现， 由此产生的性质的表征，以及活细胞中的定位研究。这项探索性研究是 预计将评估检测内源乙烯水平的可行性，并为 在细胞水平上进一步研究乙烯的产生。检测乙烯的能力 使用当前方法无法获得的空间分辨率将提供一种广泛适用的工具 报告脂质过氧化的可能性。 1