Expanding the Redox Biology Toolbox

扩展氧化还原生物学工具箱

• 批准号: 10009815
• 负责人: Xiaoli Qi
• 金额: $ 15万
• 依托单位: CHEMICAL BIOLOGY PROBES LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10009815
• 关键词: Affect; Basic Science; Biological Assay; Biological Process; Biology; Carboxylic Acids; Cell Nucleus; Cells; Charge; Chemistry; Clinical; Clinical Research; Computer software; Development; Division of Cancer Treatment and Diagnosis; Docking; Drug Screening; Electrostatics; Endoplasmic Reticulum; Equilibrium; Eukaryotic Cell; Generations; Glutathione; Goals; Half-Life; Homeostasis; Hybrids; Hydrophobicity; Laboratory Diagnosis; Lead; Legal patent; Length; Ligands; Lysosomes; Mitochondria; Monitor; National Cancer Institute; Organelles; Oxidation-Reduction; Patients; Performance; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase; Proteins; Protocols documentation; Quality Control; Reaction; Reporting; Sampling; Specificity; Sulfhydryl Compounds; Time; Tissues; Transfection; Transferase; base; commercialization; functional group; improved; interest; radioresistant; reaction rate; real time monitoring; small molecule; tool; tumor

Abstract Glutathione (GSH) is the most abundant non-protein thiol in eukaryotic cells. GSH maintains cellular redox homeostasis and is involved in numerous biological processes. These important functions are dynamically regulated by the intracellular concentration and distribution of GSH. Currently, the concentration of intracellular GSH is derived from either cell lysates or GSH-S-transferase (GST) dependent probes. These approaches, however, cannot provide information about the real-time dynamics of GSH concentration changes. We reported the first reversible reaction-based GSH probe (ThiolQuant Green) that can perform single-point quantification of GSH levels in living cells and developed the 2nd generation GSH probe—RealThiol (RT)—that allows real-time monitoring of GSH dynamics in living cells. We filed a patent on these GSH probes (WO2016025382) and have shared the RealThiol probe with >70 redox biology labs around the world. Additionally, the Division of Cancer Treatment & Diagnosis Laboratory Support at National Cancer Institute (NCI) is interested in applying our GSH probes in their drug screening assays and requested gram quantity of RT probes. The overarching goal for this Phase I project is to develop probes that can monitor the dynamics of the concentration change of GSH in real- time with subcellular specificity and to provide a complete toolset to study GSH-related redox biology. For Phase II commercialization, we will focus on both basic and clinical research. We will develop assay kits for GSH-related biology, such as ferroptosis. We will also apply our GSH probes in clinical samples to help identify patients with drug/radiation-resistant tumors.

摘要 谷胱甘肽(GSH)是真核细胞中含量最丰富的非蛋白硫醇。谷胱甘肽维持细胞氧化还原 动态平衡，参与多种生物过程。这些重要的功能是动态的 受细胞内GSH浓度和分布的调节。目前，细胞内的浓度 谷胱甘肽来源于细胞裂解产物或谷胱甘肽S转移酶依赖的探针。这些方法， 然而，不能提供有关GSH浓度变化的实时动态信息。我们报道了 第一个基于可逆反应的GSH探针(ThiolQuant Green)可以执行单点定量 活细胞中的GSH水平，并开发了第二代GSH探针-RealThiol(RT)-允许实时 活细胞内谷胱甘肽动态监测。我们为这些GSH探针申请了专利(WO2016025382)，并已 与世界各地的70个氧化还原生物实验室共享RealThiol探测器。此外，癌症部门 美国国立癌症研究所(NCI)的治疗和诊断实验室支持部门有兴趣应用我们的GSH 他们的药物筛选试验中的探针和要求的克量的RT探针。这项工作的首要目标是 一期工程是研制能够实时监测体内GSH浓度动态变化的探头。 具有亚细胞特异性的时间，并为研究GSH相关的氧化还原生物学提供了完整的工具集。适用于阶段 II商业化，我们将把重点放在基础和临床研究上。我们将开发GSH相关的检测试剂盒 生物学方面，如铁性下垂。我们还将在临床样本中应用我们的GSH探针，以帮助识别患有 耐药/耐辐射肿瘤。