IL-1Beta-Induced Retinal Iron Accumulation in Age-Related Macular Degeneration

项目总结 老年性黄斑变性(AMD)是美国最常见的不可逆性失明原因 各州。AMD的特征是光感受器及其支持细胞--视网膜色素的死亡 上皮(RPE)。Dunaef实验室之前已经证明铁在光感受器中积累 AMD患者的RPE与年龄匹配的对照组比较。随后的体外和体内工作 证明了铁的积累足以导致视网膜退化。此外，铁络合疗法 对继发于多种毒素和基因突变的视网膜变性具有保护作用 与AMD相关。然而，AMD中铁蓄积的机制尚不清楚。在这项提案中，我们 利用磁性细胞分选技术结合分子生物学技术在人和小鼠视网膜中的应用 组织来验证我们的假设：IL-1β触发视网膜铁在光感受器和视网膜色素上皮积聚，加剧 AMD。 在目标1中，我们将分离AMD和正常视网膜的细胞组分，以确定哪些细胞类型产生IL-1。 1β，以及哪些细胞类型表达IL-1β信号转导所需的受体蛋白。我们还将 测量每个细胞组份中铁转运蛋白的mRNA和蛋白水平，以确定哪些转运蛋白 导致光感受器和RPE中的视网膜铁超载。在目标2中，我们将使用细胞分类来分离 慢性神经炎小鼠模型的视网膜细胞组分。我们假设IL-1β和IL-1β 受体IL-1R1是铁积累和继发于慢性前列腺癌的铁转录组改变所必需的 炎症性刺激。我们还假设，慢性给药IL-1β，通过一种 腺相关病毒，将足以改变视网膜铁转录组和铁的积累。 这些数据将共同决定(1)IL-1β信号是否可能作为视网膜铁积聚的触发因素 (2)抑制IL-1β是否可以防止光感受器和视网膜色素上皮细胞中铁的蓄积。 这笔赠款将为我成为一名有抱负的科学家和眼科医生提供不可或缺的支持 我可以实现我的最终目标，融合临床和基础科学眼科，创造更好的 视网膜疾病的治疗方法。

PROJECT SUMMARY Age-related macular degeneration (AMD) is the most common cause of irreversible blindness in the United States. AMD is characterized by the death of photoreceptors and their supporting cells, the retinal pigment epithelium (RPE). The Dunaief lab has previously demonstrated that iron accumulates in the photoreceptors and RPE of AMD patients compared to age-matched controls. Subsequent in vitro and in vivo work demonstrated that iron accumulation is sufficient for retinal degeneration. Furthermore, iron chelation therapy is protective against retinal degeneration secondary to a diverse array of toxins and genetic mutations associated with AMD. However, the mechanism of iron accumulation in AMD is not known. In this proposal, we utilize magnetic cell sorting technology paired with molecular biology techniques in human and mouse retinal tissue to test our hypothesis: IL-1β triggers retinal iron accumulation in photoreceptors and RPE, exacerbating AMD. In Aim 1, we will isolate cell fractions from AMD and normal retinas to determine which cell-types produce IL- 1β and which cell-types express the receptor proteins necessary for IL-1β signal transduction. We will also measure mRNA and protein levels of iron transporters in each cell fraction to identify which transporters contribute to retinal iron overload in the photoreceptors and RPE. In Aim 2, we will use cell sorting to isolate retinal cell fractions from a mouse model of chronic neuroinflammation. We hypothesize that IL-1β and IL-1β receptor, IL-1R1, are necessary for iron accumulation and iron transcriptome changes secondary to a chronic inflammatory stimulus. We also hypothesize that chronic administration of IL-1β, delivered to retina using an adeno-associated virus, will be sufficient for changes in the retinal iron transcriptome and iron accumulation. Together these data will determine (1) if IL-1β signaling may serve as a trigger for retinal iron accumulation in AMD and (2) whether IL-1β inhibition can be used to prevent iron accumulation in photoreceptors and RPE. This grant will provide indispensable support towards my training as an aspiring scientist-ophthalmologist so that I can achieve my ultimate goal of fusing clinical and basic science ophthalmology to create better therapies for retinal disease.