Regulation of Glia-driven Neuroinflammation in Glaucoma

青光眼中神经胶质细胞驱动的神经炎症的调节

• 批准号: 9910408
• 负责人: Gulgun TEZEL
• 金额: $ 40万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9910408
• 关键词: American; Animal Model; Astrocytes; Attention; Autoantibodies; Autopsy; Axon; Blindness; Clinical; Comparative Study; Complement; Data; Disease; Drug Targeting; Ensure; Genetic Transcription; Glaucoma; Homeostasis; Human; Image; Immune; Individual; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Injections; Injury; Mass Spectrum Analysis; Methodology; Methods; Microglia; Microspheres; Modeling; Molecular; Molecular Analysis; Mouse Strains; Mus; Nerve Degeneration; Neuroglia; Neuronal Injury; Neurons; Ocular Hypertension; Optic Disk; Optic Nerve; Outcome; Outcome Study; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Pilot Projects; Play; Process; Production; Regulation; Retina; Role; Sampling; Site; Structure; T-Lymphocyte; Testing; Time; Transcription Coactivator; Transgenic Mice; Transgenic Organisms; Vision; Work; adaptive immune response; anterior chamber; base; cell type; chemokine; cytokine; design; event cycle; experimental study; glial activation; immunogenic; immunoregulation; inflammatory milieu; neuroinflammation; neuroprotection; neurotoxic; neurotoxicity; prevent; response; treatment strategy; viscoelasticity

PROJECT SUMMARY Recent studies of clinical/postmortem and experimental samples have accumulated evidence that supports a connection of inflammatory responses to neurodegeneration in glaucoma. Although inflammatory aspects of glaucomatous neurodegeneration are attracting increasing attention, how neuroinflammation is regulated in glaucoma, and how we can prevent neurodegenerative inflammation remain unknown. Here, we present experimental data leading us to hypothesize that NF-κB plays a key role in activating the glia-driven neuroinflammation in the glaucomatous retina and optic nerve, and that targeting this transcriptional activator mechanism is a logical strategy to provide immunomodulation and avoid secondary injury processes. With respect to intimate inter-relationship between astroglia and microglia in induction of the inflammatory/neurotoxic phenotype, NF-κB, the key transcriptional activator of inflammatory mediators, should be critical for both glial subtypes and their interaction in neuroinflammation in glaucoma. To test this hypothesis, we will comparatively study glial subtype-targeted conditional transgenic lines (in which the NF-κB canonical pathway is inactivated by drug-induced deletion of IκKβ in astroglia or microglia) and background controls with or without ocular hypertension (induced by anterior chamber microbead/viscoelastic injections). We will longitudinally analyze glial subtype-targeted transgenic effects on a diverse set of inflammatory outcomes at different sites of injury (retina, optic nerve head, and optic nerve axons). The studied outcomes will include the glial inflammatory activation phenotype and cell type-specific effects of glial subtype-targeted IκKβ deletion. Besides a preselected set of molecules, freshly isolated samples of astroglia and microglia will be analyzed by quantitative mass spectrometry to also gain high-throughput molecular information about transgenic effects on NF-κB-regulated inflammation pathways, other transcriptional targets of NF-κB, and molecules relevant to glial neurosupport functions. Since glia drive both innate and adaptive immune responses in glaucoma, additional analysis will include T-cell reactivity and autoantibody production. To determine whether transgenic inhibition of glial NF-κB (and neuroinflammation) protects neurons from immunogenic injury in mouse glaucoma, we will also analyze neuron structure (by RGC and axon counts) and function (by PERG). We expect that this project, complementing the astroglia- or microglia- targeted transgenic lines with molecular analysis of astroglia-specific and microglia-specific responses (in isolated cell type-specific samples) will allow us to determine the role of NF-κB in individual contribution and inter- relationship of astroglia and microglia in neuroinflammation (in the retina, optic nerve head, and axons), characterize specific molecular components, and value glial NF-κB as a treatment target to restore immune homeostasis and provide neuroprotection through immunomodulation in glaucoma.

项目摘要 最近对临床/死后和实验样本的研究积累了证据，支持 青光眼炎症反应与神经变性的关系虽然炎症方面的 神经退行性变引起越来越多的关注，神经炎症是如何调节的， 青光眼，以及我们如何预防神经退行性炎症仍然未知。在这里，我们介绍 实验数据使我们假设NF-κB在激活胶质细胞驱动的 神经炎症的视网膜和视神经，并针对这种转录激活剂， 机制是提供免疫调节和避免二次损伤过程的逻辑策略。相对 星形胶质细胞和小胶质细胞在诱导炎症/神经毒性方面的密切相互关系 NF-κB是炎症介质的关键转录激活因子，在神经胶质细胞和神经胶质细胞中起关键作用。 青光眼神经炎症的亚型及其相互作用。为了验证这一假设，我们将比较 研究胶质细胞亚型靶向的条件转基因系（其中NF-κB经典途径被 药物诱导的星形胶质细胞或小胶质细胞中IκKβ的缺失）和背景对照， 高血压（由前房微珠/粘弹性注射引起）。我们将纵向分析胶质细胞 亚型靶向的转基因对不同损伤部位的各种炎症结果的影响（视网膜， 视神经头和视神经轴突）。研究结果将包括胶质细胞炎症激活 胶质细胞亚型靶向IκKβ缺失的表型和细胞类型特异性效应。除了一套预选的 分子，新鲜分离的星形胶质细胞和小胶质细胞样本将通过定量质谱分析 同时获得转基因对NF-κ B调节炎症的影响的高通量分子信息 途径、NF-κB的其他转录靶点以及与胶质细胞神经支持功能相关的分子。自从胶质细胞 驱动青光眼的先天性和适应性免疫反应，额外的分析将包括T细胞反应性 和自身抗体的产生。为了确定转基因是否抑制胶质细胞NF-κB（和神经炎症） 保护神经元免受小鼠青光眼中的免疫原性损伤，我们还将分析神经元结构（通过RGC 和轴突计数）和功能（通过PERG）。我们希望这个项目，补充星形胶质细胞-或小胶质细胞- 具有星形胶质细胞特异性和小胶质细胞特异性反应的分子分析的靶向转基因系（在分离的 细胞类型特异性样本）将使我们能够确定NF-κB在个体贡献和细胞间作用中的作用。 星形胶质细胞和小胶质细胞在神经炎症中的关系（在视网膜、视神经头和轴突中）， 表征特定的分子组分，并将胶质细胞NF-κB作为恢复免疫的治疗靶点 并通过免疫调节在青光眼中提供神经保护。