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

IL-1β 诱导的年龄相关性黄斑变性中视网膜铁积累

• 批准号: 10547817
• 负责人: Jacob Khan Sterling
• 金额: $ 1.81万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2023-05-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10547817
• 关键词: Acute; Age; Age related macular degeneration; Animals; Antibodies; Aqueous Humor; Astrocytes; Autopsy; Basic Science; Blindness; Brain; CASP1 gene; Cell Fraction; Cell Separation; Cells; Cessation of life; Chronic; Clinical Sciences; DNA Sequence Alteration; Data; Dependovirus; Development; Dose; Down-Regulation; Exposure to; FDA approved; Fluorescent in Situ Hybridization; Fostering; Genes; Genetic Transcription; Goals; Grant; Hepatocyte; Homeostasis; Human; IL1R1 gene; Immunofluorescence Immunologic; In Vitro; Individual; Inflammation; Inflammatory; Interleukin-1 beta; Iron; Iron Overload; Link; Lipopolysaccharides; Macrophage; Magnetism; Maps; Measures; Messenger RNA; Microdissection; Microglia; Molecular Biology Techniques; Muller' s cell; Mus; Nerve Degeneration; Neuroglia; Normal Cell; Ophthalmologist; Ophthalmology; Oxidative Stress; Pathogenesis; Pathway interactions; Patients; Photoreceptors; Population; Production; Protein Export Pathway; Protein Import; Proteins; Proteome; Publishing; Research; Retina; Retinal Degeneration; Retinal Diseases; Rod; Saline; Scientist; Secondary to; Serum; Signal Transduction; Signaling Protein; Starvation; Stimulus; Structure of retinal pigment epithelium; Supporting Cell; Technology; Testing; Tissues; Toxin; Training; United States; Up-Regulation; Vascular Endothelial Cell; Work; cell type; cellular targeting; cytokine; epithelial injury; in vivo; in vivo Model; iron chelation therapy; macula; mouse model; neuroinflammation; neurosensory; novel therapeutic intervention; overexpression; pathogen; photoreceptor degeneration; prevent; proteostasis; receptor; response; systemic inflammatory response; transcriptome

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.

项目概要 年龄相关性黄斑变性 (AMD) 是美国不可逆性失明的最常见原因 国家。 AMD 的特点是光感受器及其支持细胞（视网膜色素）的死亡 上皮细胞（RPE）。 Dunaief 实验室此前已证明铁会积聚在光感受器中 AMD 患者的 RPE 和 RPE 与年龄匹配的对照组相比。随后的体外和体内工作 证明铁积累足以导致视网膜变性。此外，铁螯合疗法 可以预防继发于多种毒素和基因突变的视网膜变性 与 AMD 相关。然而，AMD 中铁积累的机制尚不清楚。在这个提案中，我们 在人类和小鼠视网膜中利用磁性细胞分选技术与分子生物学技术相结合 组织来检验我们的假设：IL-1β 会触发视网膜光感受器和 RPE 中的铁积累，从而加剧 AMD。 在目标 1 中，我们将从 AMD 和正常视网膜中分离细胞部分，以确定哪些细胞类型产生 IL- 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 测量每个细胞组分中铁转运蛋白的 mRNA 和蛋白质水平，以确定哪些转运蛋白 导致视网膜光感受器和 RPE 铁过载。在目标 2 中，我们将使用细胞分选来分离 来自慢性神经炎症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 对于铁积累和继发于慢性疾病的铁转录组变化是必需的。 炎症刺激。我们还假设长期施用 IL-1β，使用 腺相关病毒足以改变视网膜铁转录组和铁积累。 这些数据将共同确定 (1) IL-1β 信号传导是否可以作为视网膜铁积累的触发因素。 AMD 和 (2) IL-1β 抑制是否可用于防止光感受器和 RPE 中铁的积累。 这笔赠款将为我作为一名有抱负的科学家眼科医生的培训提供不可或缺的支持，因此 我可以实现融合临床和基础科学眼科以创造更好的最终目标 视网膜疾病的治疗。