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 视网膜小猪上皮（RPE）是一种支持健康的视觉功能的作用，长期寿命的细胞类型 数十年来。衰老的视网膜显示出在复杂疾病中被夸大的病理特征 称为年龄相关的黄斑变性（AMD）。衰老RPE的进行性变性是 地理萎缩，AMD的后期会导致视力丧失。我们最近发现了超过的丰度 源自正弦逆转录元元件（RTE）alu家族的胞质完整性DNA 地理萎缩和胞质Alu cDNA的人眼对动物的RPE高度细胞毒性 基于人类细胞的研究。这些alu cDNA是通过长期通过alu rnas的逆转录而产生的 散布的核元素1（L1）。我们还发现，Alu cDNA触发线粒体的胞质逃逸 DNA（mtDNA），并以机械不清楚的方式一起激活这两个胞质DNA协作 多方体的先天免疫反应导致细胞死亡。为了更好地了解 Alu CDNA和MTDNA及其对视网膜衰老的因果关系的结合影响，我们将（1） 在年轻，健康 老化，受AMD影响的人眼； （2）探测衰老中Alu cDNA产生的刺激和机理 通过研究L1调节剂是否影响Alu cDNA的产生和视网膜毒性； （3）阐明 由Alu cDNA触发的线粒体功能障碍和非规范炎症途径的途径 由Alu cDNA和MTDNA关节活性触发的，这些活动负责永久性细胞死亡。该提议是 响应RFA-AG-23-015，因为它将提供有关胞质Alu cDNA和mtDNA的机械见解 通过映射其序列和表达来调节老化标志的相互作用，并调节健康状态 在衰老期间，确定其生物发生的调节，并描述它们如何彼此共谋 促进衰老中的细胞功能障碍。这些研究的完成将揭示以前意外的途径 具有用于开发变革性疗法以改善视觉健康范围的治疗性相关性 认知健康范围，如阿尔茨海默氏病等疾病具有流行病学和机械特征 与AMD。