Ifi27l2a as a therapeutic target for stroke

Ifi27l2a作为中风的治疗靶点

• 批准号: 10719517
• 负责人: GAB SEOK KIM
• 金额: $ 45.98万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10719517
• 关键词: Adult; Affect; Aging; Anti-Inflammatory Agents; Attenuated; Automobile Driving; Binding; Blood - brain barrier anatomy; Brain; Brain Injuries; Brain Ischemia; Cells; Data; Disease; Encephalitis; Female; Foundations; Gene Dosage; Genes; Gliosis; Hemorrhage; Human; Immune; In Vitro; Individual; Infarction; Inflammation; Inflammatory; Inflammatory Response; Injury; Interferon alpha; Ischemic Stroke; Knockout Mice; Mass Spectrum Analysis; Matrix Metalloproteinases; Microglia; Molecular; Morphology; Mus; Neurons; Nuclear Receptors; Outcome; Pathology; Persons; Phagocytosis; Play; Primary Cell Cultures; Production; Protein Isoforms; Proteins; Proteomics; Reactive Oxygen Species; Recombinant Proteins; Recovery; Regulation; Role; Sampling; Signal Transduction; Stroke; Testing; Therapeutic; Up-Regulation; Vascular Diseases; Viral; age related neurodegeneration; aged; aging brain; attenuation; blood-brain barrier disruption; brain cell; brain endothelial cell; cell type; cytokine; disability; effectiveness testing; extracellular; functional outcomes; glial activation; human old age (65+); immune cell infiltrate; improved outcome; in vivo; ischemic injury; knock-down; male; neuroinflammation; new therapeutic target; novel; novel strategies; overexpression; post stroke; progenitor; repaired; senescence; sex; sexual dimorphism; small hairpin RNA; stroke model; stroke patient; stroke therapy; therapeutic target; transcriptomics

Abstract Microglia (MG), the resident innate immune cells of the brain, play a critical role in the inflammatory response to stroke. Stroke predominantly affects older individuals. Unfortunately, with aging, MG become increasingly dysfunctional and, in conjunction with other cell types in the brain, contribute to worse outcome following stroke. However, the role of MG in the stroke brain is not exclusively bad, as MG also contribute to repair and recovery following stroke. Thus, the selective regulation of the MG inflammatory response at specific times in the aged brain could be of great therapeutic value for stroke patients. Using single-cell transcriptomic analysis (scRNAseq), we discovered a gene (interferon alpha-inducible protein 27-like protein 2A; Ifi27l2a) that was selectively and highly upregulated in aged MG and further upregulated in MG following stroke. Our initial analysis showed a positive correlation between Ifi27l2a expression and specific sets of pro-inflammatory cytokine and senescence-related genes. These and other studies suggested that Ifi27l2a might play a critical regulatory role in promoting the inflammatory response of MG in the post-stroke brain of mice. Analysis of human brain samples also showed increased expression of IFI27L2 (human isoform) in stroke brain, enhancing the translational relevance of our findings. Next, to define the functional role of Ifi27l2a, we performed a combination of in vitro and in vivo studies, which established a compelling causative role of Ifi27l2a in promoting MG proinflammatory function and in worsening stroke injury. We also established the potential for Ifi27l2a overexpression to induce matrix metalloproteinase (MMP) expression/activity and reactive oxygen species (ROS) production, which are two driving factors in blood brain barrier (BBB) disruption. Finally, using a mouse stroke model, we showed that partial deletion of Ifi27l2a (Ifi27l2a+/- mice) led to significantly improved outcome, as evidenced by reduced infarct volume and reduced microglial activation. Together, these data provide the first direct evidence for Ifi27l2a in modulating MG functionality and brain injury following stroke and establish Ifi27l2a-dependent signaling as a new therapeutic target. Our proposed aims will test the overall hypothesis that increased MG Ifi27l2a contributes to an exaggerated inflammatory response to ischemic stroke in the aged brain and that reducing MG Ifi27l2a expression can attenuate neuroinflammation, reduce BBB disruption, and improve outcome following stroke. If successful, these studies will lay the foundation for a novel strategy based on the selective attenuation of MG pro-inflammatory signaling in the treatment of stroke or other pathologies associated with neuroinflammation.

抽象的 小胶质细胞 (MG) 是大脑中常驻的先天免疫细胞，在炎症反应中发挥着关键作用 中风。中风主要影响老年人。不幸的是，随着年龄的增长，MG 变得越来越严重 功能失调，并与大脑中的其他细胞类型结合，导致中风后更糟糕的结果。 然而，MG 在中风大脑中的作用并不完全是坏的，因为 MG 还有助于修复和恢复 中风后。因此，老年人特定时间的 MG 炎症反应的选择性调节 大脑对于中风患者可能具有巨大的治疗价值。 使用单细胞转录组分析（scRNAseq），我们发现了一个基因（干扰素α诱导蛋白） 27样蛋白2A； Ifi27l2a) 在老年 MG 中选择性地高度上调，并在老年 MG 中进一步上调 中风后的 MG。我们的初步分析显示 Ifi27l2a 表达与特异性之间呈正相关。 一组促炎细胞因子和衰老相关基因。这些和其他研究表明 Ifi27l2a可能在促进卒中后MG炎症反应中发挥关键调节作用 老鼠的大脑。对人脑样本的分析还显示 IFI27L2（人亚型）的表达增加 在中风大脑中，增强我们研究结果的转化相关性。接下来，定义Ifi27l2a的功能角色， 我们进行了体外和体内研究的结合，确定了令人信服的因果作用 Ifi27l2a 促进 MG 促炎功能和恶化中风损伤。我们还设立了 Ifi27l2a 过表达诱导基质金属蛋白酶 (MMP) 表达/活性和反应性的潜力 氧物质（ROS）的产生，这是血脑屏障（BBB）破坏的两个驱动因素。最后， 使用小鼠中风模型，我们发现部分删除 Ifi27l2a（Ifi27l2a+/- 小鼠）会显着导致 改善结果，如梗塞体积减少和小胶质细胞激活减少所证明。在一起，这些 数据为 Ifi27l2a 调节 MG 功能和中风后脑损伤提供了第一个直接证据 并建立 Ifi27l2a 依赖性信号传导作为新的治疗靶点。 我们提出的目标将检验总体假设，即增加 MG Ifi27l2a 会导致夸大的 老年大脑对缺血性中风的炎症反应，减少 MG Ifi27l2a 表达可以 减轻神经炎症、减少血脑屏障破坏并改善中风后的预后。如果成功的话，这些 研究将为基于选择性减弱 MG 促炎症因子的新策略奠定基础 治疗中风或与神经炎症相关的其他病理的信号传导。