Microflow Liquid Chromatograph for Ultrasensitive Steroid Profiling by Mass Spectrometry

用于超灵敏类固醇质谱分析的微流液相色谱仪

• 批准号: RTI-2020-00588
• 负责人: Soma, Kiran
• 金额: $ 10.02万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=693498
• 关键词: Microflow; Liquid; Chromatograph; Ultrasensitive; Steroid

Stress has major physiological effects in all vertebrates. In particular, early-life stress can “program” physiology, with enduring consequences that last into adulthood. Glucocorticoids are steroid hormones that mediate the stress response. These hormones are secreted from the adrenal glands and reach their numerous target cells via the blood. In addition, exciting new research shows that glucocorticoids are locally synthesized and metabolized within the brain, lymphoid organs, and reproductive tissues.******My NSERC Discovery Grant and Discovery Accelerator Supplement focus on the local synthesis of glucocorticoids in the brain and lymphoid organs during early development. In particular, I will examine the regulation by environmental stressors, mechanisms of action, and long-lasting effects of local glucocorticoids. Prof. Collier's Discovery Grant focuses on UDP-glucuronosyltransferase enzymes. These enzymes reduce local levels of glucocorticoids via metabolism and play critical roles in development and reproduction. Both of our projects integrate multiple techniques, including molecular biology, bioinformatics, and liquid chromatography tandem mass spectrometry (LC-MS/MS).******For both projects, it is critical to measure local levels of glucocorticoids in microdissected tissue: brain and lymphoid organs (Soma) or reproductive tissues (Collier). This key task faces technical challenges, in particular the need for specific and ultrasensitive assays. With regard to specificity, mass spectrometry is the gold standard, and our labs are pioneers in developing mass spectrometry assays for steroids. With regard to sensitivity, the steroid assays in our labs are among the most sensitive in the world. However, steroids do not ionize readily, which limits the ability to measure low amounts of steroids (. The microLC can be integrated seamlessly with our existing state-of-the-art mass spectrometers. Failure to obtain the requested equipment will greatly delay our studies on local glucocorticoids.******This equipment is urgently needed to understand how stress “gets under the skin” to affect physiology. In addition, the advances in mass spectrometry will accelerate endocrine research worldwide and equip our HQP with cutting-edge technical skills that are in high demand in academia and industry. The fundamental knowledge gained from this project will impact a variety of disciplines, including endocrinology, enzymology, physiology, neuroscience, immunology, and analytical chemistry.

压力对所有脊椎动物都有重大的生理影响。特别是，早年的压力可以“编程”生理机能，带来持久的后果，一直持续到成年。糖皮质激素是调节应激反应的类固醇激素。这些激素从肾上腺分泌，并通过血液到达众多的靶细胞。此外，令人兴奋的新研究表明，糖皮质激素在大脑、淋巴器官和生殖组织内局部合成和代谢。 *****我的 NSERC 发现补助金和发现加速器补充项目重点关注早期发育过程中糖皮质激素在大脑和淋巴器官中的局部合成。我将特别研究环境压力源的调节、作用机制以及局部糖皮质激素的长期影响。 Collier 教授的发现资助重点关注 UDP-葡萄糖醛酸基转移酶。这些酶通过新陈代谢降低糖皮质激素的局部水平，并在发育和繁殖中发挥关键作用。我们的两个项目都整合了多种技术，包括分子生物学、生物信息学和液相色谱串联质谱 (LC-MS/MS)。 *****对于这两个项目，测量显微解剖组织中糖皮质激素的局部水平至关重要：脑和淋巴器官 (Soma) 或生殖组织 (Collier)。这项关键任务面临技术挑战，特别是需要特异性和超灵敏的检测。就特异性而言，质谱是黄金标准，我们的实验室是开发类固醇质谱分析的先驱。就灵敏度而言，我们实验室的类固醇检测是世界上最敏感的。然而，类固醇不易电离，这限制了测量少量类固醇的能力（。microLC 可以与我们现有的最先进的质谱仪无缝集成。未能获得所需的设备将大大延迟我们对局部糖皮质激素的研究。********迫切需要该设备来了解压力如何“进入皮肤”影响生理机能。此外，质谱仪的进步将 加速全球内分泌研究，并为我们的总部配备学术界和工业界急需的尖端技术技能。从该项目中获得的基础知识将影响多个学科，包括内分泌学、酶学、生理学、神经科学、免疫学和分析化学。