Characterization of the role of MIF on retinal health and disease

摘要 这项研究的重点是填补有关精确分子途径的知识空白， 支持视网膜炎症和影响缺血性视网膜疾病，包括糖尿病 视网膜病、血管疾病、早产儿视网膜病和镰状细胞视网膜病。电流 治疗往往不足以防止视力丧失，并增加选择性靶向其他 炎症介质可能提供新的视力挽救疗法。我们已经确定，（1）亲- 炎性细胞因子巨噬细胞迁移抑制因子（MIF）是预防炎症的药物靶点， 视网膜脱离中视网膜胶质增生和感光细胞丧失。(2)MIF在N-甲基-D- 天冬氨酸（NMDA）损伤模型，其模拟缺血介导的视网膜兴奋性毒性; 在该模型中，MIF的药理学和遗传学抑制增加了神经元存活。(3)临床上，我们 鉴定了MIF启动子多态性与视网膜前膜形成的遗传关联。 Müller胶质细胞/星形胶质细胞（MG）是ERM的主要成分，表明MIF可以发挥作用。 在视网膜胶质细胞的病理功能中起重要作用。MIF抑制剂正在进行临床评价， 各种系统性疾病。虽然抑制MIF的促炎作用可能确实是 MIF d-DT抑制剂增强的神经元存活，我们最近的研究结果强烈表明， 也存在替代机制。MIF在Müller胶质细胞/星形胶质细胞中高度表达， 被认为是神经胶质生长因子。我们的初步数据表明，条件性抑制 MG中的MIF增强损伤期间视网膜神经元的存活并影响MG JAK/STAT 通路在此，具体目标1将检验MIF抑制促进神经元存活的假设 通过激活Müller神经胶质细胞/星形胶质细胞的gp 130/JAK/STAT信号通路，在 鸡和小鼠NMDA模型，我们将使用药理学和遗传学方法来评估 对由MIF抑制诱导的MG信号通路和神经元存活的影响。具体目标2将 检验Müller胶质细胞/星形胶质细胞MIF的条件性缺失上调了 gp 130/JAK/STAT信号通路，并增强视网膜神经元的存活。在第三阶段，我们将 开发用MIF抑制剂治疗的受损和未受损视网膜的单细胞RNA-seq数据库 和/或MIF的MG特异性遗传缺失。我们将全面评估转录 在神经胶质细胞和视网膜神经元中的单细胞分辨率的变化，这是由MIF的抑制引起的。这 研究将确定MIF和信号通路之间的重要功能关系， 视网膜损伤中的特定细胞。从了解这些知识中获得的基本知识 转录组“开关”将为未来针对关键分子途径的研究奠定基础， 可药用，副作用最小，但能够预防和恢复视网膜损伤导致的视力丧失。

Abstract This research focuses on filling gaps in knowledge about the precise molecular pathways that underpin retinal inflammation and impact crosstalk from ischemic retinal diseases, including diabetic retinopathy, vascular diseases, retinopathy of prematurity, and sickle cell retinopathy. Current treatments are often inadequate to prevent vision loss, and adding selective targeting of additional inflammatory mediators may offer new vision-saving therapies. We have identified that (1) the pro- inflammatory cytokine macrophage migration inhibitor factor (MIF) is a druggable target for preventing retinal gliosis and photoreceptor loss in retinal detachment. (2) MIF is up-regulated in the N-methyl-D- aspartic acid (NMDA) damage model which simulates ischemia-mediated retinal excitotoxicity; pharmacologic and genetic inhibition of MIF increases neuronal survival in this model. (3) Clinically we identified a genetic association of MIF promoter polymorphisms with epiretinal membrane formation. Müller glia/astrocytes (MG) are the predominant components of ERM suggesting that MIF could play an important role in the pathological function of retinal glia. MIF inhibitors are in clinical evaluation for a variety of systemic diseases. While inhibition of MIF’s pro-inflammatory effects may indeed underlie the enhanced neuronal survival from MIF d-DT inhibitors, our recent findings strongly suggest that alternative mechanisms also exist. MIF is highly expressed in the Müller glia/astrocytes and it has been hypothesized to be a glial growth factor. Our preliminary data show that conditional inhibition of MIF in the MG enhances the survival of retinal neurons during damage and affects the MG JAK/STAT pathway. Herein, Specific Aim 1 will test the hypothesis that MIF inhibition promotes neuronal survival in retinal damage by activating the gp130/JAK/STAT signaling pathway of Müller glia/astrocytes. In chick and murine NMDA models, we will use pharmacologic and genetic approaches to assess the impact on MG signaling pathways and neuronal survival induced by MIF inhibition. Specific Aim 2 will test the hypothesis that conditional deletion of Müller glia/astrocyte MIF up-regulates the gp130/JAK/STAT pathway and enhances the survival of retinal neurons. In Specific Aim 3 we will develop a single cell RNA-seq database of damaged and undamaged retina treated with MIF inhibitors and/or MG-specific genetic deletion of MIF. We will comprehensively evaluate the transcriptional changes at single-cell resolution in the glia and retinal neurons that result from inhibition of MIF. This research will define the important functional relationships between MIF and signaling pathways on specific cells during retinal damage. The fundamental knowledge gained from understanding the transcriptome ‘switch’ will set the stage for future studies targeting key molecular pathways that are druggable with minimal side effects, but able to prevent and recover visual loss from retinal damage.