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: 10221785
• 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/10221785
• 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.

项目概要/摘要 尽管神经保护策略已显示出希望，但尚无治疗方法在中风后显示出疗效。 该项目重点研究二十二烷酸 (DOC) 介质的神经保护生物活性：神经保护素 D1 (NPD1)、Resolvin D1 (RvD1) 及其组合 (NPD1+RvD1) 抗缺血和栓塞实验 中风。这些脂质介质是根据中风的发作“按需”生物合成的，以解决中风的问题 神经炎症并恢复体内平衡。我们的初步研究表明，NPD1 给药后 神经元细胞培养物中的 OGD 会修改基因簇和上游潜在主调节因子，从而减少 神经元凋亡和神经炎症，改善细胞稳态，并对基因产生有益影响 表达于缺血再灌注。此外，我们表明 DOC 可以促进神经/行为恢复， 减少梗塞面积，增加神经发生，促进缺血性中风后细胞存活。总体目标为 我们的研究旨在揭示 DOC 在 MCAo 中作用的机制。我们的中心假设是 DOC 通过在神经元保护之前靶向选择性基因簇来促进神经元和星形胶质细胞的完整性 以及由中脑星形胶质细胞衍生的神经营养细胞调节的稳态信号整合 因子 (MANF) 和无名指蛋白 146 (Iduna)。具体目标 1 将检验 DOC 监管的假设 通过调节特定基因簇，促进 MCAo 后的稳态和细胞存活生物活性。我们将定义 剂量和治疗窗，以及它们对缺血半暗带的影响，我们将确定是否 脂质介质在有或没有溶栓组织纤溶酶原激活剂的情况下对栓塞性中风具有神经保护作用。 我们从神经元培养数据中选择了基因，包括一些编码 lncRNA 的基因，并建议 通过 RT-qPCR 高通量微流体工作流程定义有和没有 DOC 的 MCAo 后它们的表达。 具体目标 2 将检验 MANF 和 Iduna 通过 DOC 整合增强丰度的假设 神经干细胞的稳态信号恢复和增殖导致神经保护。两者都是亲 针对不同神经保护机制的生存蛋白。由于 NPD1 生物合成受以下因素刺激 神经营养素，我们将探讨 DHA→DOC（NPD1、RvD1 和 NPD1+RvD1）→MANF→Iduna→ 之间的关系 通过未折叠蛋白响应促生存信号输出进行保护。 A DOC or combinations will outline 由 MANF 和 Iduna 驱动的未折叠蛋白响应的输出。由于缺血性中风 UPR 信号传导 我们将 lncRNA 定义为 UPR 的调节器和效应器，可微调 UPR 的输出 应激信号通路并确定哪些特定基因特征是 MANF 和/或 Iduna 依赖。拟议研究的科学前提是确定最有效的 DOC 或 DOC 与神经保护/神经恢复所需的靶基因簇的组合 脂质介质，这将为未来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。