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 In recent unpublished studies we have shown plasma 2-CLFA levels are elevated 40-fold in a five-subject cohort of humans exposed to chlorine at a water treatment facility near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。氯是一种主要的化学物质 在美国按批量生产。由于火车脱轨，意外发生了接触Cl2的事件 游泳池维修工人的滥用，水处理设施的事故，以及化学战。 由于Cl2既是一种化学战剂，又是一种潜在的化学物质，因此故意暴露是一个主要问题。 恐怖分子特工。需要诊断生物标记物来确定人类接触Cl2的程度和 预测结果，这最终可能导致改善治疗支持和对策。福特汽车 实验室发现，Cl2和HOCl2都针对宿主血浆蛋白原脂质，导致2-氯脂肪醛(2-CLFALD)和2-氯脂肪酸(2-CLFA)的产生。这些氯化脂肪在体内积累到强健的水平 亚致死量氯气对小鼠肺和血浆的影响。在最近未发表的研究中，我们有 显示血浆2-CLFA水平在一组五个受试者中升高了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来自于 是目前人类接触Cl2的最好的生物标志物，具有 可能被用来预测未来的结果，这可能会导致暴露后改善护理。 使用三重四极杆仪器进行LC/MS检测，对氯化脂质进行定量。这个 对LC/MS检测的依赖是使用这些实施Cl2暴露的现场分析的限制 生物标志物。拟议的研究将通过识别氯脂修饰的蛋白质来解决这个问题。 在接触Cl2后产生。未来生产氯脂修饰蛋白抗体的研究可能 然后用来开发诊断试剂盒。有人认为，氯化脂质对蛋白质具有共价修饰作用。 氯化脂肪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，它被设想为 鉴定氯化脂质修饰蛋白作为Cl2暴露的结果可能为未来提供新的靶点 开展调查以制定应对Cl2暴露的对策。有三个具体目标。 特定目标1将鉴定由2-CLFALD修饰的氨基酸基序和蛋白质。 特殊目的2将鉴定氨基酸基序和2-CLFA修饰的蛋白质。 特定目标3将从接触Cl2的小鼠的肺和血浆中鉴定氯化脂质修饰的蛋白质。 这些研究将确定氯化脂质修饰的蛋白质，并表明它们存在于Cl2的血浆中 暴露的小鼠。此外，这些拟议的R21研究有可能导致在 未来将制定改进的应对措施，并改善Cl2暴露的结果。