Chlorinated lipid modification of proteins: Biomarkers of chlorine gas exposure

蛋白质的氯化脂质修饰：氯气暴露的生物标志物

• 批准号: 10160912
• 负责人: DAVID A. FORD
• 金额: $ 18.94万
• 依托单位: SAINT LOUIS UNIVERSITY
• 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/10160912
• 关键词: Accidents; Acids; Address; Alabama; Aldehydes; Amino Acid Motifs; Amino Acids; Antibodies; Biological Assay; Biological Markers; Caring; Chemical Warfare; Chemical Warfare Agents; Chemicals; Chemistry; Chlorine; Dependence; Detection; Diagnostic Reagent Kits; Dose; Electrons; Employment; Endothelial Cells; Epithelial Cells; Exposure to; Future; Gases; Glutathione; Human; Hypochlorous Acid; Inhalation; Investigation; Lead; Letters; Lipids; Lung; Maintenance; Modification; Mus; Outcome; Oxidants; Pharmacologic Substance; Phospholipids; Plasma; Plasmalogens; Post-Translational Protein Processing; Production; Proteins; Public Health; Research Personnel; Structure; Swimming Pools; Therapeutic; Tissue Banks; Training; Transportation; Water; adduct; airway epithelium; analog; antibody detection; chlorine gas; cohort; detection assay; diagnostic biomarker; exposed human population; improved; improved outcome; instrument; liquid chromatography mass spectrometry; manufacturing process; new therapeutic target; novel; outcome prediction; oxidation; protein biomarkers; rapid detection; signature molecule; synthetic peptide; therapeutic target; water treatment

Exposure to chlorine gas (Cl2) presents a significant threat to public health. Chlorine is a leading chemical produced by volume in the US. Exposure to Cl2 has occurred as a result of train derailments, accidental misuse by swimming pool maintenance workers, accidents at water treatment facilities, and chemical warfare. Intentional exposure is a major concern since Cl2 is both a chemical warfare agent and potential chemical terror agent. Diagnostic biomarkers are needed to determine the extent of Cl2 exposure to humans and to predict outcomes, which ultimately could lead to improved therapeutic support and countermeasures. The Ford lab discovered that both Cl2 and HOCl target host plasmalogen lipids, resulting in 2-chlorofatty aldehyde (2-CLFALD) and 2-chlorofatty acid (2-CLFA) production. These chlorinated lipids accumulate to robust levels in lung and plasma of mice exposed to sub-lethal amounts of chlorine gas. In recent unpublished studies we have shown plasma 2-CLFA levels are elevated 40-fold in a five-subject cohort of humans exposed to chlorine at a water treatment facility near Birmingham, Alabama. Taken together, we have shown chlorinated lipids derived from plasmalogen oxidation are currently the best biomarkers of Cl2 exposure to humans and have the potential to be used to predict future outcomes that could lead to improved care following exposure.!Plasma chlorinated lipids are quantified using LC/MS detection employing a triple quadrupole instrument. The dependence on LC/MS detection is a limitation in implementing field analyses of Cl2 exposure using these biomarkers. The proposed studies will address this problem by identifying chlorolipid-modified proteins produced following Cl2 exposure. Future studies producing antibodies to chlorolipid-modified proteins could then be used to develop diagnostic kits. It is proposed that chlorinated lipids covalently modify proteins. The identification of specific proteins and amino acid residues modified by chlorinated lipids will be used in future studies to develop antibodies that can be used for rapid detection assays. Additionally, it is envisioned that identifying chlorinated lipid-modified proteins as a result of Cl2 exposure may provide new targets for future investigations to develop countermeasures to Cl2 exposure. There are three specific aims. Specific Aim 1 will identify amino acid motifs and proteins modified by 2-CLFALD. Specific Aim 2 will identify amino acid motifs and proteins modified by 2-CLFA. Specific Aim 3 will identify chlorinated lipid-modified proteins in lung and plasma from mice exposed to Cl2. These studies will identify chlorinated lipid-modified proteins, and show that they are present in plasma of Cl2 exposed mice. Furthermore, these proposed R21 studies have the potential to lead to new investigations in the future to develop improved countermeasures for, and improve outcomes of, Cl2 exposure.

接触氯气 (Cl2) 对公众健康构成重大威胁。氯是一种主要化学物质 在美国批量生产。因火车脱轨、意外事故而接触 Cl2 游泳池维护人员的滥用、水处理设施的事故以及化学战。 故意接触是一个主要问题，因为 Cl2 既是化学战剂，又是潜在的化学物质 恐怖分子特工。需要诊断生物标志物来确定人类接触 Cl2 的程度并 预测结果，最终可以改善治疗支持和对策。福特 实验室发现 Cl2 和 HOCl 都靶向宿主缩醛磷脂脂质，从而产生 2-氯脂肪酸 (2-CLFALD) 和 2-氯脂肪酸 (2-CLFA)。这些氯化脂质在 暴露于亚致死量氯气的小鼠的肺和血浆。在最近未发表的研究中，我们有 研究显示，在 5 名受试者中，在一定浓度下暴露于氯的人群中，血浆 2-CLFA 水平升高了 40 倍。 阿拉巴马州伯明翰附近的水处理设施。综上所述，我们已经证明了氯化脂质衍生 缩醛磷脂氧化反应是目前人类 Cl2 暴露的最佳生物标志物，并且具有 有潜力用于预测未来的结果，从而改善暴露后的护理。！血浆 使用三重四极杆仪器通过 LC/MS 检测对氯化脂质进行定量。这 对 LC/MS 检测的依赖是使用这些方法实施 Cl2 暴露现场分析的限制 生物标志物。拟议的研究将通过鉴定氯脂修饰的蛋白质来解决这个问题 暴露于 Cl2 后产生。未来的研究可能会产生针对氯脂修饰蛋白的抗体 然后用于开发诊断试剂盒。有人提出氯化脂质共价修饰蛋白质。 氯化脂质修饰的特定蛋白质和氨基酸残基的鉴定将用于 未来的研究将开发可用于快速检测分析的抗体。此外，预计 识别由于 Cl2 暴露而导致的氯化脂质修饰蛋白可能为未来提供新的目标 调查以制定针对 Cl2 暴露的对策。具体目标有三个。 具体目标 1 将鉴定经 2-CLFALD 修饰的氨基酸基序和蛋白质。 具体目标 2 将鉴定经 2-CLFA 修饰的氨基酸基序和蛋白质。 具体目标 3 将鉴定暴露于 Cl2 的小鼠肺和血浆中的氯化脂质修饰蛋白。 这些研究将鉴定氯化脂质修饰蛋白，并表明它们存在于 Cl2 血浆中 暴露的老鼠。此外，这些拟议的 R21 研究有可能引发新的研究 未来将针对 Cl2 暴露制定改进的对策并改善其结果。