Lipid imaging in Traumatic Brain Injury by high resolution GCIB-secondary ion mass spectrometry

高分辨率 GCIB 二次离子质谱法在创伤性脑损伤中的脂质成像

• 批准号: 10454918
• 负责人: Hülya Bayir
• 金额: $ 54.22万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454918
• 关键词: Achievement; Acute Brain Injuries; Adult; Anatomy; Apoptosis; Apoptotic; Area; Attention; Biochemical; Biology; Brain; Brain Injuries; Brain region; Cardiolipins; Cell Death; Cells; Cellular biology; Cessation of life; Chemistry; Child; Contusions; Craniocerebral Trauma; Detection; Development; Disease; Event; Gases; Goals; Health; Image; Imaging Techniques; Imaging technology; Individual; Inflammatory Response; Injury; Inner mitochondrial membrane; Intracranial Hypertension; Ions; Knowledge; Label; Laboratories; Lead; Leadership; Life; Lipid Biochemistry; Lipids; Macromolecular Complexes; Maps; Mass Spectrum Analysis; Mechanics; Membrane; Membrane Lipids; Methodology; Microglia; Microscopy; Mitochondria; Modeling; Molecular; Mus; Neuroglia; Neurons; Neurosciences; Organelles; Oxidation-Reduction; Pathway interactions; Pennsylvania; Phosphatidylethanolamine; Phospholipids; Physical Chemistry; Physics; Polyunsaturated Fatty Acids; Positioning Attribute; Protocols documentation; Ptosis; Refractory; Resolution; Role; Signal Transduction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spectrometry, Mass, Secondary Ion; Surface; TBI Patients; Techniques; Technology; Testing; Therapeutic; Therapy Evaluation; Time; Tissues; Traumatic Brain Injury; Universities; Weight; Work; aqueous; base; brain tissue; cell type; controlled cortical impact; design; effectiveness evaluation; high resolution imaging; improved; innovation; interdisciplinary approach; interest; invention; lipidome; lipidomics; mass spectrometric imaging; monounsaturated fat; mortality; neuropathology; new therapeutic target; novel; novel strategies; oxidation; predictive marker; prevent; programs; response; saturated fat; shift work; small molecule; spatiotemporal; submicron

Since the invention of microscopy and the initial observation of cells more than three hundred years ago, cell biology has been triumphant in detailed structural and functional characterization of intracellular organelles and macromolecular complexes. The realization that specialized bi-functional molecules, lipids, can form the aqueous interfaces of membrane structures has attracted attention to this group of intracellular compounds. Extensive biochemical studies discovered a huge diversification of lipids that could not be accommodated within a simple concept of their role as membrane building blocks. Indeed, numerous signaling functions of different lipid molecules, including membrane lipids, have been discovered. In spite of the very successful analytical work in biochemical characterization of the countless lipids, the exact intracellular topography of individual molecular species of lipids in the context of their signaling functions has not been established. The major reason for this was the lack of adequate technologies for high resolution imaging of small lipid molecules. The most recent developments of Gas Cluster Ion Beams Secondary Ion Mass Spectrometry (GCIB-SIMS) allows, for the first time, to fill this gap of fundamental knowledge in cell biology and develop a new type of microscopy – biochemical microscopy of lipids – that will create intracellular maps of individual lipids and their essential for life asymmetric distribution in biomembranes. Achievement of the goals of this innovative and paradigm shifting work will be based on highly interdisciplinary approaches and the leadership position of the three teams in their respective fields of analytical/physical chemistry of SIMS (at Pennsylvania State University, N. Winograd), lipid biochemistry/biology (at the University of Pittsburgh, V.E. Kagan), and traumatic brain injury (TBI) (at University of Pittsburgh, H. Bayır. Aim 1 will employ high-resolution GCIB-SIMS to explore molecular speciation and construct cell-specific maps of CL and PE in neuronal, glial, and microglial cells in different anatomical regions of normal mouse brain. Aim 2 will identify TBI induced molecular alterations in cardiolipin (CL) and phosphatidylethanolamine (PE) in neuronal, glial, and microglial cells using GCIB-SIMS in mouse controlled cortical impact (CCI) model. We will further identify TBI induced changes in subcellular distribution of individual CL and PE species related to the execution of apoptotic or ferroptotic programs in the respective cells. We will be particularly interested in pro-apoptotic changes in mitochondrial CL and pro-ferroptotic changes in PE. We will also examine brain tissue removed from TBI patients with refractory intracranial hypertension and brain-bank control tissue. Aim 3 will determine the utility of GCIB-SIMS imaging in assessing the effectiveness of select anti-apoptotic and anti-ferroptotic small molecule regulators in preventing cell-specific changes in CL and PE molecular speciation after TBI. Proposed studies will decode specific features of topography of individual types of lipid molecules in cells and tissues and their role in signaling functions in health and disease.

自从三百多年前显微镜的发明和对细胞的初步观察以来， 生物学在细胞内细胞器的详细结构和功能表征方面取得了胜利， 大分子复合物认识到专门的双功能分子，脂质，可以形成 膜结构的水界面引起了对这组细胞内化合物的注意。 广泛的生物化学研究发现了一个巨大的多样化的脂质， 在一个简单的概念中，它们的作用是膜构建块。事实上，许多信号功能， 已经发现了不同的脂质分子，包括膜脂质。尽管非常成功， 在无数脂质的生化特性分析工作，精确的细胞内地形， 脂质在它们的信号传导功能方面的单个分子种类还没有建立。的 主要原因是缺乏足够的技术来对小脂质进行高分辨率成像 分子。气体团簇离子束二次离子质谱的最新进展 （GCIB-SIMS）首次填补了细胞生物学基础知识的空白，并开发了一种 一种新型的显微镜-脂质的生化显微镜-将创建个体的细胞内地图 脂质及其生命必需品在生物膜中的不对称分布。实现这一目标 创新和范式转变的工作将基于高度跨学科的方法和领导 三个团队在西姆斯（宾夕法尼亚州）分析/物理化学各自领域的地位 州立大学，N。Winograd）、脂质生物化学/生物学（匹兹堡大学，V.E.卡根），以及 创伤性脑损伤（TBI）（匹兹堡大学，H。拜尔Aim 1将采用高分辨率GCIB-SIMS 探索神经元、胶质细胞和小胶质细胞中CL和PE的分子形态并构建细胞特异性图谱 正常小鼠大脑不同解剖区域的细胞。目的2：鉴定TBI诱导的分子 在神经元、神经胶质和小胶质细胞中心磷脂（CL）和磷脂酰乙醇胺（PE）的变化， GCIB-SIMS在小鼠控制的皮质撞击（CCI）模型中。我们将进一步确定TBI引起的变化， 与细胞凋亡或铁凋亡相关的单个CL和PE物质的亚细胞分布 在各自的细胞中。我们将特别感兴趣的是线粒体CL中促凋亡的变化， 和PE中的促铁代谢变化。我们还将检查从TBI患者中取出的脑组织， 颅内高压和脑库对照组织。目标3将确定GCIB-SIMS成像的实用性 在评估选择的抗细胞凋亡和抗铁凋亡小分子调节剂在 防止TBI后CL和PE分子形态的细胞特异性变化。拟议的研究将解码 细胞和组织中个别类型脂质分子的拓扑结构的具体特征及其在 在健康和疾病中的信号功能。