"Myeloid PFKFB3 in subretinal fibrosis"

“视网膜下纤维化中的骨髓 PFKFB3”

基本信息

批准号：
10342773
负责人：
Ruth B Caldwell
金额：
$ 40.03万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10342773
关键词：
6-Phosphofructo-2-kinase 6-Phosphofructokinase Anatomy Area Bone Marrow Cell Proliferation Cells Choroid Choroidal Neovascularization Cicatrix Citric Acid Cycle Complex Data Development Endothelial Cells Enzyme Activation Enzymes Exudative age-related macular degeneration Eye diseases Fibrosis Fructose Fructose-2,6-bisphosphatase Genetic Glycolysis In Vitro Lasers Measures Mediating Mesenchymal Metabolic Metabolic Pathway Metabolism Modeling Molecular Mus Myelogenous Myeloid Cells Myofibroblast Neuroglia Oxygen Pathway interactions Pharmacology Production Profibrotic signal Protein Isoforms Role Smooth Muscle Actin Staining Method Structure of retinal pigment epithelium Testing Therapeutic VLDL receptor Visual impairment cell type design epithelial to mesenchymal transition in vitro Model in vivo inhibitor knock-down macrophage novel strategies overexpression retinal angiogenesis tool treatment strategy

项目摘要

REVISED PROJECT SUMMARY Subretinal fibrosis is an end-stage fibrous plaque/disciform scar that progresses from choroidal neovascularization (CNV) during neovascular age-related macular degeneration (nAMD). Subretinal fibrosis compromises highly organized anatomical layers and tightly coordinated cellular interactions, inevitably leading to irreversible visual impairment. Current treatment for subretinal fibrosis is limited and thus, therapeutic strategies for the inhibition of subretinal fibrosis are imperative. Multiple cell types, including endothelial cells (ECs), retinal pigment epithelium (RPE) cells, macrophages, and glial cells, contribute to subretinal fibrosis by either differentiating into mesenchymal-like cells and further differentiating into α-smooth muscle actin-positive myofibroblasts and/or producing profibrotic and proinflammatory factors. However, the underlying metabolic mechanisms for these cellular and molecular activities remain poorly defined. Glycolysis is a metabolic pathway utilized by many proliferative cells. Our preliminary data show that cells in subretinal fibrotic areas are hyper-glycolytic, as evidenced by high levels of glycolytic enzymes and glycolytic regulators/activators including 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase isoform 3 (Pfkfb3), a critical enzyme for activation of glycolysis in various highly proliferative cells. Pfkfb3 catalyzes the synthesis of fructose-2,6-bisphosphate (F2, 6P2), which is the most potent allosteric activator of 6-phosphofructo-1-kinase (PFK-1), a rate-limiting enzyme for glycolysis. We have demonstrated that high levels of glycolytic enzymes including Pfkfb3 are present in the RPE/choroid complex isolated from laser-induced and spontaneous subretinal fibrosis in C57BL/6j mice and very low–density lipoprotein receptor deficient (Vldlr-/-) mice and that the area of subretinal fibrosis is markedly decreased in Pfkfb3-/+ mice. Our in vitro studies have also shown that PFKFB3/Pfkfb3 deletion in myeloid cells inhibits their transition to myofibroblast cells as well as reducing their production of proinflammatory and profibrotic factors. We hypothesize that Pfkfb3-mediated glycolysis in myeloid cells induces their transition to myofibroblasts and induces their production of profibrotic and proinflammatory factors by activating HIFs pathways, eventually leading to the development of subretinal fibrosis. To test our hypothesis, we have generated a variety of genetic mice and established mouse subretinal fibrosis models with laser-induced CNV and spontaneous CNV in Vldlr-/- mice. We will investigate the effect on subretinal fibrosis of Pfkfb3 deficiency or inhibition in myeloid cells using specific genetic and pharmacological tools with an integrated approach of in vivo and in vitro models. Our study will define the role of PFKFB3-mediated metabolism in myeloid cells in the development of subretinal fibrosis and validate inhibition of myeloid PFKFB3 as a novel strategy for the treatment of subretinal fibrosis.

修订项目摘要视网膜下纤维化是一种在新血管相关的黄斑变性（NAMD）期间从脉络膜新生血管形成（CNV）进行的终端纤维斑块/椎间盘疤痕。视网膜下纤维化会损害高度组织的解剖层和紧密协调的细胞相互作用，这不可避免地导致不可逆的视觉障碍。视网膜下纤维化的当前治疗受到限制，因此，抑制视网膜下纤维化的治疗策略必须进行。多种细胞类型，包括内皮细胞（EC），视网膜色素上皮（RPE）细胞，巨噬细胞和神经胶质细胞，通过分化为间充质样细胞并进一步分化为α-氨基肌肉肌蛋白肌蛋白阳性肌纤维蛋白肌纤维细胞和/或产生蛋白质纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维纤维的纤维化和产生的良好，并进一步分化，从而有助于产肾上腺素的纤维化。但是，这些细胞和分子活性的潜在代谢机制仍然很差。糖酵解是许多增殖细胞使用的代谢途径。我们的初步数据表明，下视网膜下纤维化区域的细胞是高糖酵解的，这是由高水平的糖酵解酶和糖酵解调节剂/活化剂（包括6-磷酸2-激酶/果糖2-激酶-2，6-磷酸酶同步酶3）的高度繁殖单元格（PFKFB3）的高度grally Encifation civitifation in Activation in Cristifation in Cristifation in Activation in Cristifation。 PFKFB3催化果糖-2,6-三磷酸盐（F2，6P2）的合成，这是6-磷酸果果-1-激酶（PFK-1）的最潜在的变构激活剂，这是一种用于糖溶解的速率酶酶。我们已经证明，在C57BL/6J小鼠中，从激光诱导的和赞助者自发的下视网膜下纤维化分离的RPE/脉络性复合物中存在高水平的糖酵解酶，以及非常低密度的脂蛋白受体缺陷（VLDLR-////sark sark sarmintial s subtinal-clibrible）（vldlr--/smartinal in Subtinal in subtribal）（vldlr-/smartinal）subtribr-subtribr（vldlr--/s）。 PFKFB3 - /+小鼠。我们的体外研究还表明，髓样细胞中的PFKFB3/PFKFB3缺失抑制其过渡到肌纤维细胞，并减少其促炎和促炎因子的产生。我们假设PFKFB3介导的髓样细胞中的糖酵解会影响它们向肌纤维细胞的过渡，并通过激活HIFS途径来诱导其产生纤维化和促炎性因子的产生，最终导致肾上腺素纤维化的发展。为了检验我们的假设，我们已经产生了各种遗传小鼠，并用激光诱导的CNV和VLDLR - / - 小鼠中的CNV建立了小鼠sbletinal纤维化模型。我们将使用特定的遗传和药物工具研究具有体内和体外模型的整合方法，研究对髓样细胞中PFKFB3缺乏症或抑制的影响。我们的研究将定义PFKFB3介导的代谢在髓样细胞中的作用，在肾上腺纤维纤维化的发展中，并验证对髓样PFKFB3的抑制作用，这是治疗肾上腺结束纤维化的新策略。