Regulators of Photoreceptor Aerobic Glycolysis in Retinal Health and Disease

项目总结/摘要 瓦尔堡效应（WE）是在肿瘤中观察到的葡萄糖向乳酸的厌氧转化增强， 视网膜细胞，即使在正常氧下。肿瘤细胞和视网膜细胞都将葡萄糖重新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 年龄相关性黄斑变性（AMD），折磨着1500万美国人， 他们的愿景合成代谢过程由糖酵解酶丙酮酸激酶M2（PKM 2）和 醛缩酶醛缩酶通过调节PKM 2活性（磷酸烯醇丙酮酸[PEP]的转化）促进抗肿瘤 丙酮酸[Pyr]），其由其磷酸化状态控制。此外，抗PKM 2和抗ALDOC 已在AMD患者视网膜中鉴定出抗体，暗示它们参与合成代谢失调 在AMD发病机制中的作用。因此，了解PKM 2和ALDOC之间的相互作用-两个关键的调解人， WE -将有助于阐明在健康视网膜中调节合成代谢过程的机制， 它们是如何在致病条件下失调的。 我们的初步研究表明，非受体酪氨酸激酶Src在体外磷酸化PKM 2。我们早先 研究表明，在视网膜中Src激活和PTP 1B活性抑制是光和视紫红质依赖性的。 我们将检验cSrc和PTP 1B调节PKM 2磷酸化的假设，从而确定 光感受器（PR）细胞中的光和视紫红质依赖性调节的机制（Aim 1）。在 目的2，我们将检验ALDOC在PR和视网膜色素中以不同方式影响PKM 2的假设。 上皮（RPE）细胞，从而确定ALDOC在RPE和PR细胞中的调节作用。目标3将测试 视网膜老化和AMD伴随PKM 2依赖性的动态变化的假说， 翻译的mRNA库，从而阐明PKM 2在调节PR和RPE基因表达中的作用 该项目的结果将是对WE介质如何调节视网膜有更深入的了解 结构、功能和可行性。这些知识对于理解公关和 RPE细胞在正常和病理条件下发挥功能并保持存活，并且与使命直接相关 美国国家眼科研究所（National Eye Institute，NIH）这些研究的成功完成将为发展 预防/延缓视网膜变性的策略。

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.