Microglia-Vascular Interaction in Alteration of Blood-Retinal Barrier in Diabetic Retinopathy

意义：糖尿病是一种全球流行病，美国有4800万人受到影响，患病率为 预计到2045年将增加到6300万。这种疾病是一个重大的公共卫生问题。糖尿病 在退伍军人健康管理局(VHA)内构成了一个重大挑战，因为估计 在其登记的患者中，糖尿病的患病率高达25%，大大高于一般患者 人口。糖尿病是第三种最常见的VHA诊断方法，占药房费用的25% 医院床位数超过170万天。糖尿病视网膜病变是糖尿病的一种微血管并发症 导致中年失明的原因。尽管使用了针对血管内皮细胞的治疗药物 生长因子(VEGF)，约三分之二的患者视力无法恢复，且作用是短暂的，需要 反复玻璃体内注射药物。理论基础：视网膜的慢性炎症在BRB中起主要作用 蚀变导致血管渗漏。我们的初步工作提供了新的证据，表明 糖尿病视网膜单核细胞内流和小胶质细胞激活。这项研究提案的目标是 确定激活的小胶质细胞分泌哪些因子，以及这些因子如何与血管相互作用 导致BRB改变的细胞。假设：糖尿病视网膜分泌物中激活的小胶质细胞 影响周细胞-内皮细胞相互作用导致BRB改变的因素。目标：1) 糖尿病患者小胶质细胞耗竭对视网膜血管和结构的影响 视网膜病变，2a)通过分析视网膜病变的核心转录信号来识别和优先处理候选基因 糖尿病动物模型中视网膜小胶质细胞激活与血-视网膜屏障改变的关系。 2b)确认“小胶质细胞候选基因”对内皮细胞通透性和血视网膜的作用 糖尿病的屏障改变和3)测试功能验证的候选基因的可药性 在细胞培养模型和糖尿病动物模型中使用非专利小分子文库。设计：In 这项研究计划，下一代序列(NGS)技术将被用来研究分子 糖尿病患者BRB改变的基础。这项研究将解决一个关键的科学差距，通过确定 血管内皮生长因子以外的改变BRB的分子介质。相关性：这些发现可能有助于开发 1型和2型糖尿病中针对这些分子的新生物标志物和改进的治疗策略 对于那些对抗血管内皮生长因子药物反应不佳的患者，可能会防止视力丧失。我们的方法是 创新是因为这项提议引入了一项创新战略，其目标是稳定 糖尿病小胶质细胞功能障碍的初始阶段，这是糖尿病发展的关键上游事件 黄斑水肿。

Significance: Diabetes is a global epidemic with 48 million people affected in USA, and the prevalence is expected to increase to 63 million by 2045. The disease represents a significant public health problem. Diabetes within the Veterans Health Administration (VHA) poses a significant challenge because the estimated prevalence of diabetes among its enrolled patients is as high as 25%, substantially higher than the general population. Diabetes is the third most common VHA diagnosis and accounted for 25% of pharmacy costs and over 1.7 million hospital bed days. Diabetic retinopathy, a microvascular complication of diabetes is the leading cause of vision loss in middle-aged adults. Despite the use of therapeutics targeting the vascular endothelial growth factor (VEGF), about two thirds of patients do not recover vision, and the effect is transient, needing repeated intravitreal injections of drugs. Rationale: Chronic inflammation in retina plays a major role in BRB alteration resulting in vascular leakage. Our preliminary work presents novel evidence that there is increased influx of monocytes and activation of microglia in the retina in diabetes. The goal of this research proposal is to determine what factors are secreted by the activated microglia and how those factors interact with vascular cells leading to alteration of the BRB. Hypothesis: Activated microglia in the diabetic retina secrete factors that affect pericyte-endothelial interactions leading to BRB alteration. Aims: 1) Determine effect of microglia depletion on the retinal vasculature and structure in the context of diabetic retinopathy, 2a) Identify and prioritize “candidate genes” by analyzing the core transcriptional signatures of activated retinal microglia in an animal model of diabetes in relationship to blood-retinal barrier alteration. 2b) Validate the role of “microglial candidate genes” on endothelial cell permeability and blood retinal barrier alteration in diabetes and 3) Test the druggability of the functionally validated candidate genes using an off-patent small molecule library in cell culture models and a diabetic animal model. Design: In this research proposal, next generation sequence (NGS) technology will be utilized to investigate the molecular basis of alteration of the BRB in diabetes. This research will address a critical scientific gap by identifying molecular mediators beyond VEGF that alter the BRB. Relevance: The findings may help in development of novel biomarkers and improved therapeutic strategies targeting these molecules in both type 1 and 2 diabetes patients that may prevent vision loss in those who poorly respond to anti-VEGF drugs. Our approach is innovative because this proposal introduces an innovative strategy that targets the mechanisms of stabilizing the initial step of microglia dysfunction in diabetes, a critical upstream event for the development of diabetic macular edema.