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.

项目摘要/摘要 与年龄相关的黄斑变性（AMD）代表发达国家失明的主要原因，并且 大量研究已致力于探索原因和潜在治疗方法。而线粒体 已经发现对AMD的发展至关重要，对确切途径的了解很少 线粒体会影响核基因表达和疾病进展。我们已经开发了 新型的蒙上膜软骨细胞体模型，使我们能够产生残留的色素上皮细胞系 核，但具有不同AMD和年龄匹配的正常受试者的线粒体。使用这些气缸，我们的 实验室发现，引入AMD线粒体会导致细胞凋亡，氧化应激和 扩展的细胞活力。目前，我们正在探索AMD患者的线粒体如何引起这些 细胞因子细胞系的急剧变化，以及如何调节这种影响以改善细胞健康。一 我们投资的潜在途径是MicroRNA的监管。我们的初步cybrid数据 证明AMD线粒体的引入会导致七个microRNA的表达改变具有功能 与AMD病理学有关。随后的工作集中于两个过表达的目标下调 MicroRNA存在于AMD周期（miRNA 135b-5p和miRNA 148a-3p）中。 miRNA 135b-下调 5p导致与凋亡和血管生成相关的基因表达降低，而下调下调 miRNA 148A-3P的表达水平增加了与线粒体生物发生和 活性氧的产生减少。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方法的方法。基于这些 初步数据，我们对该赠款的中心假设是AMD线粒体（a）暴露的细胞改变了 microRNA表达以及（b）通过靶向抑制调节这些失调的microRNA水平 或过表达会影响基因表达，细胞健康和血管生成的体外模型。更远 了解microRNA在AMD模型中的作用对各种其他 有线粒体功能障碍的疾病，包括阿尔茨海默氏症和帕金森氏病。