RPE Messengers, Transcription and Photoreceptor Renewal

RPE 信使、转录和光感受器更新

• 批准号: 10418615
• 负责人: Nicolas G. Bazan
• 金额: $ 44.79万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-03-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10418615
• 关键词: Affect; Age related macular degeneration; Animal Model; Awareness; Cell Death; Cell Maintenance; Cell physiology; Cells; Disease model; Docosahexaenoic Acids; Eicosapentaenoic Acid; Environment; Essential Fatty Acids; Event; Eye; Family member; Fatty Acids; Functional disorder; Future; G-Protein-Coupled Receptors; Gene Cluster; Gene Proteins; Genes; Genetic Transcription; Genomics; Goals; Grant; Homeostasis; Human; Inflammatory; Investigation; Knowledge; Lead; Lentivirus; Life; Light; Lipids; Mediating; Mediator of activation protein; Membrane; Metabolism; Methods; Molecular; Molecular Genetics; Neuroglia; Omega-3 Fatty Acids; Outcome; Oxygen Consumption; Pathway interactions; Phenotype; Phosphorylation; Photoreceptors; Polyunsaturated Fatty Acids; Proteins; Rejuvenation; Retina; Retinal Degeneration; Retinoids; Role; Signal Pathway; Signal Transduction; Site; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Structure of retinal pigment epithelium; Techniques; Testing; Time; Very Long Chain Fatty Acid; Vision; adiponectin; base; cell injury; cell type; gene network; inherited retinal degeneration; lipid mediator; lipidomics; molecular imaging; neuroprotectin D1; novel; oxidative damage; peroxidation; photoreceptor cell inner segment; pigment epithelium-derived factor; programs; receptor; targeted treatment; translational impact; uptake

Project Summary/Abstract Despite their adverse environment, the question of how photoreceptor cells (PRC) and retinal pigment epithelial cells (RPEC) remain functionally unchanged for decades in healthy eyes remains to be answered. This grant program is focused on the molecular cascade by which bioactive derivatives of the essential fatty acid family member docosahexaenoic acid (DHA) induce effectors through GPC receptors to sustain protection of PRC and RPEC function. We have shown that the synthesis of one DHA-derived mediator, Neuroprotectin D1 (NPD1), is activated at the onset of homeostatic disruptions, and we have recently discovered a second set of mediators, elovanoids (ELVs; derived from precursors made by ELOVL4), that are critical in the cascade. Our next goal is to answer two important questions: Question 1. What mechanisms get DHA to the right places at the right times in order to give rise to the lipid mediators? Hypothesis 1 – Several retinal cells [RPEC, PRC, and Müller/glia (MG)] facilitate DHA uptake, retention, and metabolism via multiple pathways to support the synthesis of the homeostatic lipid mediators sustaining PRC integrity. Question 2. How does the lipid-mediator signaling sustain cellular integrity? Hypothesis 2 – NPD1 and ELV are synthetized in the RPEC to sustain function of these cells in a hostile environment. These mediators are synthesized on demand when homeostatic disruption occurs which, in turn, activates counteracting signaling against oxidative-cell damage. Mechanisms include GPC receptor-mediated gene clusters, which regulate the expression of specific proteins that promote homeostasis and maintenance of cell integrity. Aim 1 tests the prediction that specific mechanisms and pathways facilitate the uptake and metabolism of DHA in RPECs, MGs and PRCs to sustain PRC renewal. Aim 2 tests the prediction that NPD1 and ELV elicit their bioactivity by targeting specific G-protein-coupled receptors. Aim 3 tests the prediction that NPD1 and ELV contribute to RPEC and PRC homeostasis by targeting gene clusters. Genes will be identified using inducible lentivirus constructs, AdipoR1 global KO, cell-type specific AdipoR1 conditional KO and MFRP (rd6) animal models and single-cell genomics techniques. Additional methods include LC–MS/MS- based lipidomic analysis, MALDI molecular imaging and human primary RPECs. The outcomes of this fundamental investigation will provide new information and understanding about DHA acquisition and retention, as well as of the basic molecular and genetic signals and mechanisms required to sustain retinal function. The results of this study may also define the parameters for future studies on the pathophysiology and targets for treatment of age-related macular degeneration as well as of inherited retinal degenerations.

项目摘要/摘要 尽管环境不利，但感光细胞（PRC）和视网膜色素上皮的问题 细胞（RPEC）在健康的眼睛中保持数十年的功能保持不变。这笔赠款 程序的重点是分子级联 成员Docosahexaenoic Acid（DHA）通过GPC受体影响效果，以维持中国的保护和 RPEC功能。我们已经表明，一个DHA衍生的介体神经保护素D1（NPD1）的合成是 在稳态中断时被激活，我们最近发现了第二组调解人， Elovanoids（Elvs;源自Elovl4的前体），在级联反应中至关重要。我们的下一个目标是 回答两个重要的问题：问题1。哪些机制在正确的时间将DHA带到正确的位置 为了引起脂质介质？假设1 - 几个残留细胞[RPEC，PRC和Müller/Glia （MG）]通过多种途径促进DHA的吸收，保留和代谢，以支持合成 维持PRC完整性的稳态脂质介质。问题2。脂质介导器信号如何维持 细胞完整性？假设2 - NPD1和ELV在RPEC中合成以维持这些细胞的功能 在敌对的环境中。当体内稳态中断时，这些介体是按需综合的 反过来，它激活了针对氧化细胞损伤的应对信号传导。机制包括GPC 受体介导的基因簇，它调节促进稳态的特定蛋白的表达 和细胞完整性的维护。 AIM 1测试了特定机制和途径促进的预测 DHA在RPEC，MGS和PRC中的摄取和代谢，以维持Prc Renewal。 AIM 2测试预测 NPD1和ELV通过靶向特定的G蛋白偶联受体来引起其生物活性。 AIM 3测试 预测NPD1和ELV通过靶向基因簇促进了RPEC和PRC稳态。基因会 使用诱导式慢病毒构建体ADIPOR1 Global KO，细胞型特异性ADIPOR1条件KO鉴定 和MFRP（RD6）动物模型和单细胞基因组学技术。其他方法包括LC – MS/MS- 基于脂质组分析，MALDI分子成像和人类原发性RPEC。结果的结果 基本投资将提供有关DHA获取和保留的新信息和理解， 以及维持剩余功能所需的基本分子和遗传信号和机制。这 这项研究的结果还可以定义有关病理生理学的未来研究的参数和目标 治疗与年龄相关的黄斑变性以及遗传的残余变性。