AMD Mitochondria Modulate Expression of microRNA 135b-5p and 148a-3p in RPE Cybrids: Implications for Age-related Macular Degeneration

AMD 线粒体调节 RPE Cybrids 中 microRNA 135b-5p 和 148a-3p 的表达：对年龄相关性黄斑变性的影响

• 批准号: 10597239
• 负责人: Vladimir Jivkov Kefalov
• 金额: $ 19.63万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10597239
• 关键词: Affect; Age; Age related macular degeneration; Aging; Alzheimer' s Disease; Apoptosis; Bioenergetics; Biogenesis; Blindness; Blood Vessels; Cell Culture Techniques; Cell Death; Cell Line; Cell Nucleus; Cell Survival; Cells; Coculture Techniques; Cytoplasm; Data; Dermal; Development; Disease; Disease Progression; Down-Regulation; Effectiveness; Endothelial Cells; Exhibits; Fibroblasts; Foundations; Funding; Future; Gene Expression; Genes; Genus Hippocampus; Goals; Grant; Growth; Health; Human; In Vitro; Inflammation; Inflammatory; Laboratories; Lead; Length; Measures; Methods; MicroRNAs; Mitochondria; Modeling; Morphology; Nonexudative age-related macular degeneration; Nuclear; Oxidative Stress; Parkinson Disease; Pathologic; Pathology; Pathway interactions; Patients; Pattern; Production; Proteins; Quantitative Reverse Transcriptase PCR; RNA; Reactive Oxygen Species; Regulation; Research; Role; Signal Transduction; Structure of retinal pigment epithelium; Umbilical vein; Vascular Endothelial Growth Factors; Western Blotting; Work; angiogenesis; cytokine; imager; improved; in vitro Model; inhibitor; interest; metabolomics; microRNA biomarkers; mimicry; mitochondrial dysfunction; neuroprotection; novel; overexpression; therapeutic miRNA; therapeutic target

Project Summary/Abstract Age-related Macular Degeneration (AMD) represents a major cause of blindness in the developed world, and significant research has been devoted to exploring the causes and potential treatments. While the mitochondria have been found to be of critical importance to the development of AMD, less is known about the exact pathways through which mitochondria influence nuclear gene expression and disease progression. We have developed a novel transmitochondrial cybrid model that allows us to generate retinal pigment epithelial cell lines with identical nuclei, but with mitochondria from different AMD and age-matched normal subjects. Using these cybrids, our laboratory has discovered that introduction of AMD mitochondria causes cellular apoptosis, oxidative stress and decreased cell viability. Presently, we are exploring how the mitochondria of AMD patients could cause these dramatic changes in cybrid cell lines, and how this influence could be modulated to improve cellular health. One of the potential pathways that we have investigated is the regulation of microRNA. Our preliminary cybrid data show that introduction of AMD mitochondria causes altered expression of seven microRNA with functions relevant to AMD pathology. Subsequent work has focused on the targeted downregulation of two overexpressed microRNA present in the AMD cybrids (miRNA 135b-5p and miRNA 148a-3p). Downregulation of miRNA 135b- 5p leads to decreased expression of genes associated with apoptosis and angiogenesis, while downregulation of miRNA 148a-3p results in increased expression levels of genes associated with mitochondrial biogenesis and decreased reactive oxygen species production. These findings demonstrate the effectiveness of microRNA modulation as a method for improvement of cellular health and potential treatment. Building upon these preliminary data, our central hypotheses for this grant are that cells with AMD mitochondria (a) exhibit altered microRNA expression and that (b) modulation of these dysregulated microRNA levels through targeted inhibition or overexpression will influence gene expression, cellular health and in vitro models of angiogenesis. Further understanding of the role of microRNA in the AMD model has a significant potential impact for a variety of other diseases with mitochondrial dysfunction, including Alzheimer’s and Parkinson’s diseases.

项目概要/摘要 年龄相关性黄斑变性 (AMD) 是发达国家失明的主要原因， 重要的研究致力于探索线粒体的原因和潜在的治疗方法。 已被发现对 AMD 的发展至关重要，但对其确切途径知之甚少 我们开发了一种线粒体通过其影响核基因表达和疾病进展的方法。 新型传输软骨细胞混合模型使我们能够生成具有相同特征的视网膜色素上皮细胞系 细胞核，但具有来自不同 AMD 和年龄匹配的正常受试者的线粒体。 实验室发现AMD线粒体的引入会导致细胞凋亡、氧化应激和 目前，我们正在探索AMD患者的线粒体如何导致这些现象。 细胞系的巨大变化，以及如何调节这种影响以改善细胞健康。 我们研究的潜在途径之一是 microRNA 的调控，我们的初步 cybrid 数据。 表明 AMD 线粒体的引入导致七种具有功能的 microRNA 的表达发生改变 与 AMD 病理学相关的后续工作重点是两种过度表达的靶向下调。 AMD cybrid 中存在 microRNA（miRNA 135b-5p 和 miRNA 148a-3p）。miRNA 135b- 下调。 5p 导致与细胞凋亡和血管生成相关的基因表达减少，同时下调 miRNA 148a-3p 的增加导致与线粒体生物合成相关的基因表达水平增加 这些发现证明了 microRNA 的有效性。 调节作为改善细胞健康和潜在治疗的方法。 初步数据显示，我们对于这项资助的中心假设是，具有 AMD 线粒体 (a) 的细胞表现出改变 microRNA 表达以及 (b) 通过靶向抑制调节这些失调的 microRNA 水平 或过度表达将影响基因表达、细胞健康和血管生成的体外模型。 了解 microRNA 在 AMD 模型中的作用对于许多其他领域具有重大的潜在影响 线粒体功能障碍的疾病，包括阿尔茨海默病和帕金森病。