Cytosolic SINE retrotransposable element cDNA and mitochondrial DNA in aging retina

衰老视网膜中的胞质 SINE 逆转录转座元件 cDNA 和线粒体 DNA

• 批准号: 10722062
• 负责人: Jayakrishna Ambati
• 金额: $ 62.67万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10722062
• 关键词: Affect; Age related macular degeneration; Aging; Alu Elements; Alzheimer' s Disease; Animals; Binding; Biogenesis; Blindness; Cell Death; Cells; Cessation of life; Chromatin; Cognitive; Collaborations; Complementary DNA; Complex; Cytoplasm; DNA; Data; Disease; Elements; Epidemiology; Event; Evolution; Eye; Family; Foundations; Functional disorder; Future; Generations; Genetic; Guanosine; Human; IL18 gene; Inflammaging; Inflammasome; Inflammation; Inflammatory; Innate Immune Response; Interleukin-1 beta; Joints; Knowledge; Libraries; Maps; Mediating; Medical; Mitochondria; Mitochondrial DNA; Molecular; Mus; Nuclear; Pathogenesis; Pathologic; Pathology; Pathway interactions; Production; Property; RNA; RNA-Directed DNA Polymerase; Regulation; Retina; Retrotransposon; Reverse Transcription; Role; Signal Transduction; Sterility; Stimulus; Structure of retinal pigment epithelium; Testing; Therapeutic; Tissues; Toll-like receptors; Toxic effect; Vision; Visual; Voltage-Dependent Anion Channel; age effect; age related; aged; cell type; cytotoxic; cytotoxicity; geographic atrophy; healthspan; human disease; improved; insight; intercellular communication; micronucleus; mitochondrial dysfunction; next generation sequencing; novel; retinal toxicity; sensor; tissue degeneration

PROJECT SUMMARY The retinal pigmented epithelium (RPE) is an amitotic, long-lived cell type that supports healthy visual function for numerous decades. The aging retina displays pathologic features that are exaggerated in a complex disease called age-related macular degeneration (AMD). Progressive degeneration of the aging RPE is a hallmark of geographic atrophy, a late-stage of AMD that causes vision loss. We recently discovered an excess abundance of cytosolic complementary DNA derived from the SINE retrotransposon element (RTE) Alu family in the RPE of human eyes with geographic atrophy and that cytosolic Alu cDNAs are highly cytotoxic to the RPE in animal and human cell-based studies. These Alu cDNAs are produced via reverse transcription of Alu RNAs by the long- interspersed nuclear element-1 (L1). We also found that Alu cDNAs trigger the cytosolic escape of mitochondrial DNA (mtDNA), and together these two cytosolic DNAs collaborate, in mechanistically unclear ways, to activate a multifaceted innate immune response that results in cell death. To gain a better understanding of the combinatory impact of Alu cDNAs and mtDNA and their causal contribution to aging in the retina, we will (1) create temporal, spatial, and sequence topographic maps of Alu cDNA subfamilies and mtDNA in young, healthy aged, and AMD-affected human eyes; (2) probe the stimuli for and mechanisms of Alu cDNA generation in aging by investigating whether L1 regulators influence Alu cDNA production and retinal toxicity; and (3) elucidate the pathway of mitochondrial dysfunction triggered by Alu cDNA and the noncanonical inflammatory pathways triggered by Alu cDNA and mtDNA joint activity that are responsible for retinal cell death. This proposal is responsive to RFA-AG-23-015 as it will provide mechanistic insights into how cytosolic Alu cDNAs and mtDNA mediate interactions of aging hallmarks and modulate healthspan by mapping their sequence and expression during aging, determining the regulation of their biogenesis, and delineating how they conspire with each other to promote cellular dysfunction in aging. Completion of these studies will reveal previously unexplored pathways with therapeutic relevance for developing transformative therapies to improve visual healthspan and potentially cognitive healthspan, as conditions such as Alzheimer’s disease share epidemiological and mechanistic features with AMD.

项目摘要 视网膜色素上皮细胞（RPE）是一种无丝分裂、寿命长的细胞类型，支持健康的视觉功能 几十年了老化的视网膜显示病理特征是夸大了一个复杂的疾病 年龄相关性黄斑变性（AMD）老化RPE的进行性变性是视网膜色素变性的标志。 地图状萎缩，一种导致视力丧失的AMD晚期。我们最近发现了大量的 来自SINE逆转录转座子元件（RTE）Alu家族的胞质互补DNA在RPE中的表达， 具有地图状萎缩人眼和细胞溶质Alu cDNA对动物中的RPE具有高度细胞毒性， 人类细胞研究。这些Alu cDNA是通过长链RNA逆转录Alu RNA而产生的。 散布的核元件-1（L1）。我们还发现，Alu cDNAs触发线粒体的胞质逃逸， 线粒体DNA和这两种胞质DNA以机制不明的方式协同激活 一种导致细胞死亡的多方面先天免疫反应。为了更好地了解 Alu cDNA和mtDNA的组合影响及其对视网膜衰老的因果作用，我们将（1） 创建年轻、健康和健康人的Alu cDNA亚家族和mtDNA的时间、空间和序列地形图， （2）探讨Alu cDNA在衰老过程中的刺激因素和机制 通过研究L1调节子是否影响Alu cDNA的产生和视网膜毒性;（3）阐明 Alu cDNA触发的线粒体功能障碍通路与非经典炎症通路 由Alu cDNA和mtDNA联合活性触发，导致视网膜细胞死亡。这项建议是 响应于RFA-AG-23-015，因为它将提供关于细胞溶质Alu cDNA和mtDNA 介导衰老标志的相互作用，并通过绘制它们的序列和表达来调节健康寿命 在衰老过程中，确定它们的生物发生的调节，并描绘它们如何相互共谋 促进衰老过程中的细胞功能障碍。这些研究的完成将揭示以前未探索的途径 具有治疗相关性，可用于开发变革性疗法，以改善视觉健康， 认知健康跨度，如阿尔茨海默病等疾病的流行病学和机制特征 关于AMD