Oxidative Lipidomics in Pediatric Traumatic Brain Injury

氧化脂质组学在小儿创伤性脑损伤中的应用

• 批准号: 10844023
• 负责人: Hülya Bayir
• 金额: $ 33.61万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10844023
• 关键词: Acute; Acyltransferase; Adult; Affinity; Anti-Inflammatory Agents; Antioxidants; Apoptotic; Arachidonate 15-Lipoxygenase; Arachidonic Acids; Biochemical Reaction; Brain; Brain Injuries; Cardiolipins; Cause of Death; Cessation of life; Child; Childhood Injury; Chronic; Clinical Trials; Coenzyme A Ligases; Collaborations; Complex; Data; Endoplasmic Reticulum; Enzymes; Esterification; Event; Family member; Free Radical Scavengers; Free Radicals; Funding; Generations; Glutathione; Hospitalization; Hydrogen Peroxide; Hydrolysis; Hydroxides; Image; Induction of Apoptosis; Inflammatory Response; Injury; Innovative Therapy; Iron; Link; Lipid Peroxidation; Lipoxygenase Inhibitors; Lysophosphatidylcholines; Mechanics; Membrane; Mitochondria; Morbidity - disease rate; National Center for Advancing Translational Sciences; Nature; Neurological outcome; Neurons; Oxidation-Reduction; Oxidative Stress; Pathogenicity; Pathway interactions; Pattern; Peroxidases; Phosphatidylethanolamine; Phosphatidylethanolamine Binding Protein; Phospholipase; Phospholipids; Proteins; Rattus; Reduced Glutathione; Role; Scaffolding Protein; Signal Transduction; Source; Stretching; Substrate Specificity; Sulfhydryl Compounds; System; TBI treatment; Technology; Testing; Therapeutic; Therapeutic Uses; Translations; Trauma; Traumatic Brain Injury; United States National Institutes of Health; Work; adduct; baicalein; catalyst; cognitive function; controlled cortical impact; cytochrome c; design; drug discovery; experimental study; glutathione peroxidase; improved outcome; inhibitor; lipidomics; liquid chromatography mass spectrometry; mortality; neuron apoptosis; neuron loss; neuroprotection; new therapeutic target; novel therapeutics; oxidation; pediatric traumatic brain injury; peroxidation; postnatal; preference; preservation; prevent; protein complex; small molecule; small molecule inhibitor; standard care; targeted treatment

Each year in the US, severe TBI in children results in ∼7400 deaths and 60 000 hospitalizations. Fifty percent of surviving children with severe TBI have poor neurological outcome at six months. Severe TBI in children is thus a critical problem in desperate need of impactful therapies. Free radicals and oxidative stress have been uniformly accepted as universal pathogenic mechanisms of TBI prompting therapeutic use of antioxidants. Invariably, clinical trials of non-specific free radical scavengers/antioxidants failed. This suggests that true sources and mechanisms of TBI redox disbalance remain undefined, and represent a potential therapeutic opportunity. During the previous funding period we showed that lipid peroxidation after TBI in immature brain occurs as a result of controlled enzymatic reactions. We discovered that peroxidation of mitochondrial phospholipid cardiolipin (CL) represents a required stage of neuronal apoptosis after TBI in postnatal day (PND) 17 rats. We identified cytochrome c as a catalyst of CL peroxidation and showed that a mitochondria- targeted inhibitor of CL peroxidation suppressed TBI-induced apoptosis and preserved cognitive function in PND17 rats. Our latest work identified highly selective oxidation of arachidonic acid (AA) containing phosphatidylethanolamines (PE) by 15 lipoxygenase (15LOX) to be causative to ferroptosis. We discovered that PE binding protein 1 (PEBP1) complexes with 15LOX and changes its substrate specificity from free AA to AA esterified into PE to generate hydroperoxy-AA-PE death signals. Normally hydroperoxy-AA-PE are eliminated by combined action of glutathione peroxidase 4 (GPX4)/glutathione (GSH). Immature brain has lower GSH levels and GPX activity vs adult brain thus could be more vulnerable to ferroptosis upon injury. Indeed our preliminary data show that TBI leads to marked increase in expression and activity of 15LOX, and accumulation of oxidized AA-PE in PND17 rat brain. Furthermore, preliminary data indicate that inhibition of AA-PE oxidation suppresses TBI-induced neuronal death and preserves cognitive function. Thus, we hypothesize that generation of oxidized AA-PE by 15LOX/PEBP1 complex leads to neuronal death and represents a new target for drug discovery leading to innovative therapies in pediatric TBI. We propose to test our hypothesis in three Specific Aims. Aim 1 will determine the degree, spatial and temporal pattern of 15LOX/PEBP1 complex formation and AA-PE oxidation after TBI. Aim 2 will investigate the mechanisms of AA-PE oxidation in TBI-induced neuronal death. Aim 3 will design and investigate the mechanism of action and neuroprotective potential of small-molecule regulators of 15LOX and 15LOX/PEBP1 activity in TBI. These studies will employ powerful lipidomics and oxidative lipidomics technology to provide important mechanistic information on the role of PE oxidation in neuronal ferroptosis after pediatric TBI. The ability to selectively modulate PE oxidation, a critical early event in the mechanism of ferroptosis, could lead to targeted therapies for TBI and ultimately improve outcome for children after brain injury.

在美国，每年严重的儿童创伤性脑损伤会导致约 7,400 人死亡和 6 万人住院治疗。百分之五十 患有严重 TBI 的幸存儿童在六个月时神经系统结果不佳。儿童严重 TBI 是 因此，这是一个迫切需要有效治疗的关键问题。自由基和氧化应激已被 普遍认为 TBI 的普遍致病机制促使抗氧化剂的治疗使用。 非特异性自由基清除剂/抗氧化剂的临床试验总是失败。这表明，真实的 TBI 氧化还原失衡的来源和机制仍不清楚，但代表了一种潜在的治疗方法 机会。在之前的资助期间，我们发现未成熟大脑中 TBI 后的脂质过氧化 发生是受控酶促反应的结果。我们发现线粒体的过氧化 磷脂心磷脂 (CL) 代表出生后 TBI 后神经元凋亡的必要阶段 (PND)17只大鼠。我们确定细胞色素 c 是 CL 过氧化的催化剂，并表明线粒体 CL过氧化的靶向抑制剂抑制了TBI诱导的细胞凋亡并保留了认知功能 PND17 大鼠。我们的最新工作确定了含有花生四烯酸 (AA) 的高选择性氧化 15 脂氧合酶 (15LOX) 产生磷脂酰乙醇胺 (PE)，导致铁死亡。我们发现 PE 结合蛋白 1 (PEBP1) 与 15LOX 复合，并将其底物特异性从游离 AA 改变为 AA酯化成PE产生氢过氧-AA-PE死亡信号。通常氢过氧-AA-PE是 通过谷胱甘肽过氧化物酶 4 (GPX4)/谷胱甘肽 (GSH) 的联合作用消除。大脑不成熟有 与成人大脑相比，GSH 水平和 GPX 活性较低，因此在受伤时更容易出现铁死亡。 事实上，我们的初步数据表明，TBI 导致 15LOX 的表达和活性显着增加，并且 PND17 大鼠脑中氧化 AA-PE 的积累。此外，初步数据表明，抑制 AA-PE 氧化可抑制 TBI 诱导的神经元死亡并保留认知功能。因此，我们 假设 15LOX/PEBP1 复合物生成氧化 AA-PE 导致神经元死亡 代表了药物发现的新目标，从而导致儿科 TBI 的创新疗法。我们建议测试 我们在三个具体目标中的假设。目标 1 将确定的程度、空间和时间模式 TBI 后 15LOX/PEBP1 复合物的形成和 AA-PE 氧化。目标 2 将研究其机制 TBI 诱导的神经元死亡中的 AA-PE 氧化。目标 3 将设计和研究作用机制 15LOX 和 15LOX/PEBP1 活性的小分子调节剂在 TBI 中的神经保护潜力。这些 研究将采用强大的脂质组学和氧化脂质组学技术来提供重要的机制 关于 PE 氧化在儿科 TBI 后神经元铁死亡中的作用的信息。选择性的能力 调节 PE 氧化是铁死亡机制中的一个关键早期事件，可能会导致靶向治疗 治疗 TBI 并最终改善脑损伤后儿童的预后。

项目成果

Integrated -omics approach reveals persistent DNA damage rewires lipid metabolism and histone hyperacetylation via MYS-1/Tip60.

• DOI: 10.1126/sciadv.abl6083
• 发表时间: 2022-02-18
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Hamsanathan S;Anthonymuthu T;Han S;Shinglot H;Siefken E;Sims A;Sen P;Pepper HL;Snyder NW;Bayir H;Kagan V;Gurkar AU
• 通讯作者: Gurkar AU

Guidelines for measuring reactive oxygen species and oxidative damage in cells and in vivo.

• DOI: 10.1038/s42255-022-00591-z
• 发表时间: 2022-06
• 期刊: NATURE METABOLISM
• 影响因子: 20.8
• 作者: Murphy, Michael P.;Bayir, Hulya;Belousov, Vsevolod;Chang, Christopher J.;Davies, Kelvin J. A.;Davies, Michael J.;Dick, Tobias P.;Finkel, Toren;Forman, Henry J.;Janssen-Heininger, Yvonne;Gems, David;Kagan, Valerian E.;Kalyanaraman, Balaraman;Larsson, Nils-Goran;Milne, Ginger L.;Nystrom, Thomas;Poulsen, Henrik E.;Radi, Rafael;Van Remmen, Holly;Schumacker, Paul T.;Thornalley, Paul J.;Toyokuni, Shinya;Winterbourn, Christine C.;Yin, Huiyong;Halliwell, Barry
• 通讯作者: Halliwell, Barry

Aiming for the target: Mitochondrial drug delivery in traumatic brain injury.

瞄准目标：创伤性脑损伤中的线粒体药物递送。

• DOI: 10.1016/j.neuropharm.2018.07.014
• 发表时间: 2019-03
• 期刊: Neuropharmacology
• 影响因子: 4.7
• 作者: Lamade AM;Kenny EM;Anthonymuthu TS;Soysal E;Clark RSB;Kagan VE;Bayır H
• 通讯作者: Bayır H

Imaging mass spectrometry of diversified cardiolipin molecular species in the brain.

• DOI: 10.1021/ac5011876
• 发表时间: 2014-07-01
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Amoscato, A. A.;Sparvero, L. J.;He, R. R.;Watkins, S.;Bayir, H.;Kagan, V. E.
• 通讯作者: Kagan, V. E.

Genetic re-engineering of polyunsaturated phospholipid profile of Saccharomyces cerevisiae identifies a novel role for Cld1 in mitigating the effects of cardiolipin peroxidation.

• DOI: 10.1016/j.bbalip.2018.06.016
• 发表时间: 2018-10
• 期刊: Biochimica et biophysica acta. Molecular and cell biology of lipids
• 作者: Lou W;Ting HC;Reynolds CA;Tyurina YY;Tyurin VA;Li Y;Ji J;Yu W;Liang Z;Stoyanovsky DA;Anthonymuthu TS;Frasso MA;Wipf P;Greenberger JS;Bayır H;Kagan VE;Greenberg ML
• 通讯作者: Greenberg ML