Docosanoids modulate homeostasis and cell survival after ischemic stroke

二十二烷酸调节缺血性中风后的体内平衡和细胞存活

• 批准号: 10395594
• 负责人: Nicolas G. Bazan
• 金额: $ 32.87万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-15 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10395594
• 关键词: Alteplase; Anabolism; Astrocytes; Behavior assessment; Behavioral; Bioinformatics; Brain; Cell Culture Techniques; Cell Death; Cell Survival; Cells; Cerebral Ischemia; Clinical Research; Computer software; DNA Repair; Data; Development; Docosahexaenoic Acids; Dose; Experimental Models; Fostering; Future; Gene Cluster; Genes; Glucose; Glutamates; Goals; Homeostasis; Infarction; Inflammation; Intervention; Ischemia; Ischemic Brain Injury; Ischemic Penumbra; Ischemic Stroke; Knockout Mice; Knowledge; Link; Longevity; Magnetic Resonance Imaging; Mediating; Mediator of activation protein; Microfluidics; Molecular; Molecular Genetics; N-Methyl-D-Aspartate Receptors; Nervous system structure; Neurologic; Neurologic Deficit; Neurons; Omega-3 Fatty Acids; Output; Oxygen; Pathway Analysis; Phenotype; Play; Polymers; Property; Proteins; Rattus; Recovery; Reperfusion Therapy; Ring Finger Domain; Role; Signal Pathway; Signal Transduction; Stress; Stroke; Testing; Therapeutic; Up-Regulation; Western Blotting; Work; aged; cell motility; cerebral ischemic injury; deprivation; effective therapy; embolic stroke; excitotoxicity; genetic signature; improved; lipid mediator; nerve stem cell; neuroblast; neurogenesis; neuroinflammation; neuron apoptosis; neuroprotectin D1; neuroprotection; neurorestoration; neurotrophic factor; post stroke; recruit; response; restoration; stem cells; stroke model; stroke outcome; stroke therapy; thromboembolic stroke; thrombolysis; translational impact

PROJECT SUMMARY / ABSTRACT Although neuroprotective strategies have shown promise, no treatment has demonstrated efficacy after stroke. This project focuses on the neuroprotective bioactivity of docosanoid (DOC) mediators: Neuroprotectin D1 (NPD1), Resolvin D1 (RvD1), and their combination (NPD1+RvD1) against ischemic and embolic experimental stroke. These lipid mediators are biosynthesized “on demand” in response to the onset of stroke to resolve neuroinflammation and restore homeostasis. Our preliminary studies show that administration of NPD1 after OGD in neuronal cell cultures modifies clusters of genes and upstream potential master regulators that decrease neuronal apoptosis and neuroinflammation, improve cell homeostasis, and that beneficially impact genes expressed in ischemia-reperfusion. In addition, we show that DOC provide neurological/behavioral recovery, reduce infarct size, increase neurogenesis, and promote cell survival after ischemic stroke. The overall goal of our studies is to uncover a mechanistic understanding of DOC action in MCAo. Our central hypothesis is that DOC foster neuronal and astrocyte integrity by targeting selective gene clusters preceding neuronal protection and by the homeostatic signaling integration regulated by the mesencephalic astrocyte-derived neurotrophic factor (MANF) and by the ring finger protein 146 (Iduna). Specific aim 1 will test the hypothesis that DOC regulate pro-homeostatic and cell survival bioactivity after MCAo by modulating specific gene clusters. We will define the doses and therapeutic window, as well as their effect on the ischemic penumbra and we will define whether the lipid mediators are neuroprotective in embolic stroke with or without tissue plasminogen activator of thrombolysis. We selected genes from our data on neuronal cultures, including, some encoding lncRNAs, and propose to define by RT-qPCR high-throughput microfluidics workflow their expression after MCAo with and without DOC. Specific aim 2 will test the hypothesis that MANF and Iduna enhanced abundance by DOC integrates homeostatic signaling restoration and proliferation of neural stem cells leading to neuroprotection. Both are pro- survival proteins that target different neuroprotective mechanisms. Since NPD1 biosynthesis is stimulated by neurotrophins, we will explore the relationship DHA→DOC (NPD1, RvD1 and NPD1+RvD1) →MANF→Iduna→ protection by unfolded protein response pro-survival signaling outputs. A DOC or combinations will outline outputs of the unfolded protein response driven by MANF and Iduna. Since ischemic stroke engages UPR signaling we will define lncRNAs as regulators and effectors of UPR that fine-tune the output of the stress signaling pathways and identify also which specific gene signatures are MANF and or Iduna dependent. The scientific premise of the proposed studies is that identification of the most effective DOC or combination of DOC to target gene clusters necessary for neuroprotection/neurorestoration modulated by the lipid mediators which will provide the basis for future clinical studies on potential interventions to reduce the immediate and long-term consequences of stroke.

项目摘要 /摘要 尽管神经保护策略已经显示出希望，但尚未表现出中风后的效率。 该项目重点介绍了二十烷酸（DOC）介质的神经保护性生物活性：神经保护素D1 （NPD1），Resolvin D1（RVD1）及其组合（NPD1+RVD1）针对缺血和栓塞实验 中风。这些脂质介质是根据中风开始的生物合成的“按需” 神经炎症并恢复体内平衡。我们的初步研究表明，NPD1的给药 神经元细胞培养物的OGD修饰剂基因和上游潜在的主调节群的簇减少 神经元凋亡和神经炎症，改善细胞稳态，并有益地影响基因 在缺血 - 重新灌注中表达。此外，我们表明DOC提供神经/行为恢复， 减少梗塞大小，增加神经发生并促进缺血性中风后的细胞存活。总体目标 我们的研究是揭示对MCAO中DOC动作的机械理解。我们的中心假设是 DOC FOSTER神经元和星形胶质细胞完整性通过针对神经元保护之前的选择性基因簇 并由脑脑星形胶质细胞衍生的神经营养性调节的体内稳态信号传导整合 因子（manf）和环手指蛋白146（iduna）。特定目标1将检验DOC调节的假设 MCAO后，促室内和细胞存活生物活性通过调节特定基因簇。我们将定义 剂量和理论窗口及其对缺血性半月的影响，我们将定义是否定义 脂质介质在栓塞中具有神经保护作用，有或没有组织纤溶酶原激活剂的溶栓激活剂。 我们从我们的神经元培养数据的数据中选择了基因，包括一些编码lncrnas的基因，以及提出的建议。 通过RT-QPCR高通量微流体的定义，在MCAO之后使用和不使用DOC。 特定目标2将检验以下假设：MANF和IDUNA通过DOC集成增强了丰度 神经元干细胞的稳态信号恢复和增殖导致神经保护。两者都是亲 靶向不同神经保护机制的生存蛋白。由于NPD1生物合成被刺激 神经营养蛋白，我们将探索关系DHA→DOC（NPD1，RVD1和NPD1+RVD1）→MANF→IDUNA→ 通过展开的蛋白质反应促生存信号传导输出保护。文档或组合将概述 由MANF和IDUNA驱动的展开蛋白质反应的输出。由于缺血性中风参与 UPR信号我们将将lncrnas定义为调节器和UPR的影响，以微调输出 应力信号通路并还确定哪些特定基因签名是manf和iDuna 依赖。拟议研究的科学前提是，识别最有效的文档或 DOC与靶基因簇结合了由神经保护/神经抑制所必需的 脂质介质将为未来的潜在干预措施的临床研究提供基础，以减少 中风的直接和长期后果。