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）是大脑的固有免疫细胞，在炎症反应中起着至关重要的作用 中风。中风主要影响老年人。不幸的是，随着衰老，MG越来越多 功能障碍，并与大脑中的其他细胞类型结合，导致中风后结果较差。 但是，MG在中风大脑中的作用并不是完全不好，因为MG也有助于修复和恢复 下风。因此，在老年人的特定时间对Mg炎症反应的选择性调节 对于中风患者，大脑可能具有很高的治疗价值。 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类蛋白2a; IFI27L2A）在老年毫克中有选择性和高度上调，并进一步上调 中风后毫克。我们的初步分析显示IFI27L2A表达与特定之间存在正相关。 促炎性细胞因子和衰老相关基因的集合。这些和其他研究表明 IFI27L2A可能在促进MG的炎症后反应中起关键的调节作用 小鼠的大脑。人脑样品的分析还显示出IFI27L2（人类同工型）的表达增加 在中风大脑中，增强了我们发现的转化相关性。接下来，要定义IFI27L2A的功能作用， 我们进行了体外和体内研究的结合，该研究确立了令人信服的因果关系的作用 IFI27L2A在促进MG促炎功能和中风损伤中。我们还建立了 IFI27L2A过表达的潜力诱导基质金属蛋白酶（MMP）表达/活性和反应性 氧（ROS）产生，这是血脑屏障（BBB）破坏的两个驱动因素。最后， 我们使用鼠标中风模型，表明IFI27L2A（IFI27L2A +/-小鼠）的部分缺失导致显着 改善的预后，梗塞体积减少和小胶质细胞激活证明了结果。在一起，这些 数据提供了第一个直接证据，证明IFI27L2A调节中风后MG功能和脑损伤 并确定IFI27L2A依赖性信号传导作为新的治疗靶标。 我们提出的目标将检验总体假设，即增加MG IFI27L2A有助于夸大 老年大脑中缺血性中风的炎症反应，并且降低MG IFI27L2A表达可以 减弱神经炎症，减少BBB破坏并改善中风后的预后。如果成功，这些 研究将为基于MG促炎性的选择性衰减而为新的策略奠定基础 与神经炎症相关的中风或其他病理治疗中的信号传导。