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The Warburg Effect and Diabetic Retinopathy

瓦尔堡效应和糖尿病视网膜病变

基本信息

批准号：
10635331
负责人：
Ahmed S Ibrahim
金额：
$ 38.5万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10635331
关键词：
5&apos -AMP-activated protein kinase Acceleration Aging Angiogenic Switch Animals Benchmarking Bioenergetics Bioinformatics Blindness Blood Vessels CRISPR/Cas technology Cell Proliferation Cells Cellular Metabolic Process Complication Data Development Diabetes Mellitus Diabetic Retinopathy Diabetic mouse Disease Dose Endoplasmic Reticulum Endothelial Cells Endothelium Enzymes Extravasation Fright Functional disorder Gene Expression Glucose Glycolysis Goals Human Hyperglycemia Hypoxia Hypoxia Inducible Factor Inositol Knockout Mice Knowledge Lasers Lipids Mediating Mediator Metabolic Metabolism Mission Mitochondria Modeling National Eye Institute Neuroglia Neurons Oxidative Phosphorylation Oxygen Pathology Pathway Analysis Pathway interactions Patients Phenotype Phosphotransferases Production Protein Biosynthesis Proteins Research Personnel Retina Retinal Neovascularization Ribonucleases Risk Factors Role Sampling Signal Transduction Streptozocin Testing Tissues Tube Visual Acuity Warburg Effect angiogenesis bevacizumab cancer cell conditional knockout diabetic endoplasmic reticulum stress in vitro Model in vivo inhibitor insight mitochondrial dysfunction neovascularization neurovascular neurovascular unit novel nucleotide metabolism pharmacologic prevent proliferative diabetic retinopathy response sensor success

项目摘要

Project Summary/Abstract Retinal neovascularization (RNV) is a debilitating complication of advanced diabetic retinopathy, which despite the use of anti-VEGF and laser treatments continues to cause blindness. Less is known as to why RNV develops only after patients have had diabetes for decades. Although endothelial cell (EC) angiogenic activation is a hallmark of this transition, how ECs adapt their metabolism to sustain such activation remains a significant gap in our knowledge. Our long-term goal is to determine the bioenergetic mechanisms of RNV. The Warburg effect is a metabolic shift from mitochondrial oxidative phosphorylation (OxPhos) to hyperglycolysis that was first found in cancer cells. This metabolic shift not only produces ATP faster than OxPhos, albeit less efficiently, but also provides precursors required for lipid, protein, and nucleotide synthesis during cell proliferation. Recently the Warburg effect was rediscovered as a key contributor in various endothelial-related diseases; however, its role in diabetic retinopathy is not well-defined. In this application, the overall objective(s) are to define the role of the Warburg effect in diabetic retinopathy and to identify its underlying mechanisms. Here, we propose that multiple hits are needed to cooperatively alter EC metabolism to fulfill biosynthetic demands of transforming a quiescent EC into an angiogenic cell. Tissue hypoxia is the most common risk factor associated with advanced diabetic retinopathy. Thus, our central hypothesis is that diabetes primes quiescent ECs to be angiogenic (first hit) and that hypoxia (second hit) is necessary for angiogenic switch via the metabolic adaptation of the Warburg effect. Aim1 will test the hypothesis that persistent activation of the energy sensor, AMP-activated protein kinase (AMPK) sustains the Warburg effect to mediate EC angiogenic activation. Our approach is to use a two-hit model of diabetes and hypoxia in AMPKα1 endothelial-specific conditional knockout (AMPKα1End-/-) mice and in AMPKα1 silenced human retinal ECs (HRECs) to achieve this aim. We will also use vitreous samples from patients with proliferative diabetic retinopathy to test the correlation between the development of RNV and the Warburg effect- associated metabolites. Aim2 will investigate the role of endoplasmic reticulum (ER) stress in mediating the Warburg effect-induced EC angiogenic activation. We hypothesize that activation of Inositol-requiring enzyme (IRE)1, a unique ER-stress sensor protein with kinase and RNase activities, is a key mediator for the Warburg effect-induced EC angiogenic activation. We will use CRISPR/Cas9 to test the effect of inhibiting downstream signaling of IRE1 kinase and RNase activities on the Warburg effect-induced EC angiogenic activation. We will also test our hypothesis in vivo using a two-hit model of hypoxia and diabetes and selective pharmacological inhibitors. Overall, this new-investigator initiated R01 capitalizes on the interdisciplinary expertise of a biochemist, a mitochondrial biologist, an ER biologist, and a clinician to use a novel two-hit model of advanced diabetic retinopathy to gain mechanistic insights into the bioenergetic basis of RNV. Understanding the role of the Warburg effect will reveal novel targets in the treatment of diabetic retinopathy.

项目总结/摘要视网膜新生血管（RNV）是晚期糖尿病视网膜病变的一种使人衰弱的并发症，尽管抗VEGF和激光治疗的使用继续导致失明。关于RNV为什么会发展，只有在患者患有糖尿病几十年后才能这样做。虽然内皮细胞（EC）血管生成激活是一个重要因素，作为这种转变的一个标志，内皮细胞如何适应它们的代谢以维持这种激活仍然是一个重大的空白在我们的知识。我们的长期目标是确定RNV的生物能量机制。瓦尔堡效应是一种从线粒体氧化磷酸化（OxPhos）到糖酵解过度的代谢转变，在癌细胞中。这种代谢转变不仅比OxPhos更快地产生ATP，尽管效率较低，而且提供细胞增殖过程中脂质、蛋白质和核苷酸合成所需的前体。最近瓦尔堡效应被重新发现为各种内皮相关疾病的关键因素;然而，糖尿病性视网膜病变的发病机制尚不明确。在本申请中，总体目标是定义糖尿病视网膜病变中的瓦尔堡效应，并确定其潜在机制。在此，我们建议，需要多个命中来协同改变EC代谢，以满足转化EC的生物合成需求。静止EC转化为血管生成细胞。组织缺氧是与晚期胰腺癌相关的最常见的危险因素。糖尿病视网膜病变因此，我们的中心假设是糖尿病使静止的EC成为血管生成细胞（首先是血管生成细胞）。 Hit）和缺氧（第二次Hit）是通过瓦尔堡代谢适应进行血管生成转换所必需的效果Aim 1将检验能量传感器AMP激活蛋白激酶持续激活的假设，（AMPK）维持瓦尔堡效应以介导EC血管生成激活。我们的方法是使用两次打击模型 AMPK α 1内皮特异性条件性敲除（AMPK α 1End-/-）小鼠和AMPK α 1 沉默人视网膜EC（HREC）以实现这一目的。我们还将使用玻璃体样本，增殖性糖尿病视网膜病变，以测试RNV的发展和瓦尔堡效应之间的相关性- 相关代谢物。Aim2将研究内质网（ER）应激在介导细胞凋亡中的作用。瓦尔堡效应诱导的EC血管生成激活。我们假设肌醇需要酶的激活（IRE）1是一种独特的具有激酶和RNA酶活性的内质网应激传感蛋白，是瓦尔堡效应的关键介质效应诱导的EC血管生成激活。我们将使用CRISPR/Cas9来测试抑制下游的效果。 IRE1激酶和RNA酶活性对瓦尔堡效应诱导的EC血管生成激活的信号传导。我们将还使用缺氧和糖尿病的两次打击模型和选择性药理学方法在体内测试我们的假设。抑制剂的总的来说，这个新的研究者发起的R01利用了一个跨学科的专业知识，生物化学家，线粒体生物学家，ER生物学家和临床医生使用一种新的先进的两次打击模型。糖尿病视网膜病变，以获得对RNV的生物能量基础的机械见解。理解的作用瓦尔堡效应将揭示治疗糖尿病视网膜病变的新靶点。