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 In recent unpublished studies we have shown plasma 2-CLFA levels are elevated 40-fold in a five-subject cohort of humans exposed to chlorine at a water treatment facility near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）对公共卫生构成了重大威胁。氯是领先的化学物质 由美国的数量产生。由于火车出轨，意外而发生的CL2暴露 游泳池维护人员滥用，水处理设施中的事故和化学战。 故意暴露是一个主要问题，因为CL2既是化学战剂，又是潜在的化学物质 恐怖分子。需要诊断生物标志物来确定CL2暴露于人类的程度和 预测结果，最终可能会改善治疗支持和对策。福特 LAB发现CL2和HOCL靶宿主血浆脂质脂质，导致2-氯氟醛（2-Clfald）和2-氯氟脂蛋白酸（2-CLFA）产生。这些氯化脂质在 暴露于亚致死量的氯气的小鼠的肺和血浆。在最近未发表的研究中，我们有 显示的血浆2-CLFA水平在暴露于氯的五个受试者队列中升高了40倍 阿拉巴马州伯明翰附近的水处理设施。总之，我们显示了衍生的氯化脂质 目前，从血浆原成因氧化是CL2暴露于人类的最佳生物标志物，并具有 潜力用于预测未来结果，这些结果可能会导致暴露后改善护理。 使用三倍四极杆仪器使用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。拟议的研究将通过鉴定叶绿素改性蛋白来解决此问题 在CL2暴露后产生。未来产生抗叶绿素改性蛋白抗体的研究可能 然后用于开发诊断套件。建议氯化脂质共价修改蛋白质。 通过氯化脂质修饰的特定蛋白质和氨基酸残基的鉴定将用于 未来的研究开发可用于快速检测测定的抗体。另外，它设想 由于CL2暴露而识别氯化脂质改性蛋白可能会为将来提供新的目标 调查以发展与CL2暴露的对策。有三个特定的目标。 特定的目标1将鉴定由2-Clfald修饰的氨基酸基序和蛋白质。 特定的目标2将鉴定由2-CLFA修饰的氨基酸基序和蛋白质。 特定的目标3将鉴定肺中氯化脂质改性的蛋白质和暴露于CL2的小鼠的血浆。 这些研究将鉴定氯化脂质改性蛋白，并表明它们存在于Cl2的血浆中 裸露的小鼠。此外，这些拟议的R21研究有可能导致在 未来可以改善CL2暴露的对策并改善结果。