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.

项目总结/摘要 虽然神经保护策略已经显示出希望，但没有治疗在中风后表现出有效性。 本项目的重点是神经保护生物活性的docosanoid（DOC）介质：神经保护素D1 （NPD 1）、Resolvin D1（RvD 1）和它们的组合（NPD 1 + RvD 1）抗缺血性和栓塞性实验 中风这些脂质介质是生物合成的“需求”，以应对中风的发作，以解决 神经炎症和恢复稳态。我们的初步研究表明，NPD 1的管理后， 神经元细胞培养物中的OGD改变了基因簇和上游潜在的主调节因子， 神经元凋亡和神经炎症，改善细胞稳态，并有益地影响基因 在缺血再灌注中表达。此外，我们表明，DOC提供神经/行为恢复， 减少梗死面积，增加神经发生，促进缺血性卒中后细胞存活。的总目标 我们的研究旨在揭示DOC在MCAo中作用的机理。我们的核心假设是， DOC通过在神经元保护之前靶向选择性基因簇促进神经元和星形胶质细胞的完整性 以及中脑星形胶质细胞源性神经营养因子调节的稳态信号整合 环指蛋白146（Iduna）。具体目标1将检验DOC调节 通过调节特定的基因簇，在MCAo后具有促稳态和细胞存活的生物活性。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组织纤溶酶原激活剂溶栓的栓塞性卒中中具有神经保护作用。 我们从神经元培养的数据中选择了基因，包括一些编码lncRNA的基因，并提出 通过RT-qPCR高通量微流体工作流程定义它们在有和没有DOC的MCAo后的表达。 具体目标2将检验MANF和Iduna通过DOC整合提高丰度的假设 稳态信号传导恢复和神经干细胞增殖导致神经保护。两者都是亲-- 针对不同神经保护机制的生存蛋白。由于NPD 1的生物合成受到 神经营养因子，我们将探讨DHA→DOC（NPD 1，RvD 1和NPD 1 + RvD 1）→MANF→Iduna→ 通过未折叠的蛋白质应答促存活信号输出进行保护。DOC或组合将概述 由MANF和Iduna驱动的未折叠蛋白质响应的输出。由于缺血性中风 我们将lncRNA定义为UPR的调节因子和效应因子， 应激信号通路，并鉴定哪些特定基因标记是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与靶向基因簇的组合，这些基因簇是由神经保护/神经恢复调节所必需的 脂质介质，这将为未来的潜在干预措施，以减少临床研究的基础。 中风的直接和长期后果。