Regulators of Photoreceptor Aerobic Glycolysis in Retinal Health and Disease

视网膜健康和疾病中光感受器有氧糖酵解的调节因子

• 批准号: 10717825
• 负责人: Raju VS Rajala
• 金额: $ 42.93万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10717825
• 关键词: Ablation; Affect; Affinity Chromatography; Age related macular degeneration; Aging; Aldehyde-Lyases; Aldolase C; American; Anabolism; Antibodies; Antioxidants; Cell physiology; Cells; Complex; DNA; Disease; Enzymes; Gene Expression; Gene Expression Profile; Glucose; Glycolysis; Goals; Health; In Vitro; Interphase Cell; Knockout Mice; Knowledge; Label; Light; Lipids; Mediating; Mediator; Messenger RNA; Metabolic; Metabolism; Mission; Molecular; Mus; National Eye Institute; Outcome; Pathogenesis; Pathogenicity; Pathologic; Pathology; Patients; Phosphoenolpyruvate; Phosphorylation; Photoreceptors; Process; Protein Dephosphorylation; Protein Tyrosine Kinase; Proteins; Publications; Pyruvate; Pyruvate Kinase; RNA; Regulation; Retina; Retinal Degeneration; Rhodopsin; Ribosomes; Rod; Role; Signal Transduction; Structure; Structure of retinal pigment epithelium; Techniques; Testing; Transcription Coactivator; Translating; Tyrosine Phosphorylation; United States National Institutes of Health; Vision; Warburg Effect; Work; aerobic glycolysis; aged; design; experimental study; innovation; knock-down; lipid biosynthesis; mouse model; normoxia; novel; novel therapeutics; prevent; protein tyrosine phosphatase 1B; retinal rods; src-Family Kinases; tumor

PROJECT SUMMARY/ABSTRACT The Warburg Effect (WE) is the enhanced anaerobic conversion of glucose to lactate observed in tumor and retinal cells, even in normoxia. Both tumor and retinal cells repurpose glucose for anabolic biosynthesis of lipid, protein, RNA/DNA, and antioxidant molecules. In the retina, dysregulation of anabolic processes is associated with age-related macular degeneration (AMD), which afflicts 15 million Americans with devastating effects on their vision. Anabolic processes are regulated by the glycolytic enzymes pyruvate kinase M2 (PKM2) and aldolase. Aldolase promotes anabolism by regulating PKM2 activity (conversion of phosphoenolpyruvate [PEP] to pyruvate [Pyr]), which is controlled by its phosphorylation status. Furthermore, anti-PKM2 and anti-ALDOC antibodies have been identified in AMD patient retinas, implicating their involvement in anabolic dysregulation in AMD pathogenesis. Thus, understanding the interplay between PKM2 and ALDOC - two key mediators of the WE - will help elucidate the mechanisms by which anabolic processes are regulated in healthy retinas and how they are dysregulated in pathogenic conditions. Our preliminary studies show that non-receptor tyrosine kinase Src phosphorylates PKM2 in vitro. Our earlier studies show that in the retina Src activation and PTP1B activity inhibition are light- and rhodopsin-dependent. We will test the hypothesis that cSrc and PTP1B regulate PKM2 phosphorylation, thereby determining the mechanism of light- and rhodopsin-dependent regulation of anabolism in photoreceptor (PR) cells (Aim 1). In Aim 2, we will test the hypothesis that ALDOC affects PKM2 in diverse ways in the PR and retinal pigment epithelial (RPE) cells, thereby determining the regulatory role of ALDOC in RPE and PR cells. Aim 3 will test the hypothesis that retinal aging and AMD are accompanied by a PKM2-dependent dynamic change in actively translated mRNA repertoire, thereby elucidating the role of PKM2 in regulating PR and RPE gene expression The outcome of this project will be a deeper understanding of how mediators of the WE regulate retina structure, functions, and viability. This knowledge is fundamentally important for understanding how PR and RPE cells function and stay alive in normal and pathologic conditions, and are directly relevant to the mission of the National Eye Institute, NIH. Successful completion of these studies will pave the path for developing strategies to prevent/delay retinal degeneration.

项目摘要/摘要 Warburg效应（We）是在肿瘤和 视网膜细胞，即使在常氧中也是如此。肿瘤和视网膜细胞都重新利用葡萄糖，用于脂质的合成代谢生物合成， 蛋白质，RNA/DNA和抗氧化剂分子。在视网膜中，合成代谢过程的失调与 与年龄相关的黄斑变性（AMD），折磨了1500万美国人对 他们的愿景。合成代谢过程由糖粉酶丙酮酸激酶M2（PKM2）和 藻酶。通过调节PKM2活性（磷酸烯醇丙酮酸[PEP]转化[PEP] 丙酮酸[pyr]），该）由其磷酸化状态控制。此外，抗PKM2和抗aldoc 在AMD患者视网膜中已经鉴定出抗体，暗示其参与合成代谢失调 在AMD发病机理中。因此，了解PKM2和ALDOC之间的相互作用 - 我们将有助于阐明在健康的视网膜中调节合成代谢过程的机制 在致病条件下它们的失调方式。 我们的初步研究表明，非受体酪氨酸激酶SRC在体外磷酸化PKM2。我们的早期 研究表明，在视网膜中，SRC激活和PTP1B活性抑制作用是光和视紫红质依赖性的。 我们将测试CSRC和PTP1B调节PKM2磷酸化的假设，从而确定 光感受器（PR）细胞中代谢的光和视紫红质依赖性调节的机理（AIM 1）。在 AIM 2，我们将检验以下假设：ALDOC在PR和视网膜色素中以各种方式影响PKM2 上皮（RPE）细胞，从而确定ALDOC在RPE和PR细胞中的调节作用。 AIM 3将测试 视网膜老化和AMD伴随着PKM2依赖性动态变化的假设 翻译了mRNA曲目，从而阐明了PKM2在调节PR和RPE基因表达中的作用 该项目的结果将更深入地了解我们调节视网膜的调节器 结构，功能和生存能力。这些知识对于了解公关和如何如何 RPE细胞在正常和病理条件下起作用并保持活力，并且与任务直接相关 NIH国家眼科研究所。这些研究的成功完成将为发展铺平道路 预防/延迟视网膜变性的策略。