Myeloid glycolysis in pathological ocular angiogenesis

病理性眼血管生成中的髓样糖酵解

基本信息

批准号：
10673058
负责人：
Ruth B Caldwell
金额：
$ 45.28万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10673058
关键词：
6-Phosphofructo-2-kinase 6-Phosphofructokinase Adult Age Age related macular degeneration Angiogenic Factor Aorta Area Biological Assay Blindness Blood Vessels Bone Marrow Breeding Cell surface Cells Choroid Coculture Techniques Conditioned Culture Media Consensus Critical Pathways Data Development Elderly Endothelial Cells Endothelium Enzyme Activation Enzymes Exhibits Eye diseases Fructose Fructose-2,6-bisphosphatase Genetic Glycolysis Growth HIF1A gene Histone Acetylation Hypoxia Hypoxia Inducible Factor In Vitro Individual Inflammatory Laser injury Lesion Leukocytes Macrophage Measures Mediating Messenger RNA Metabolism Methods Microglia Molecular Mus Myelogenous Myeloid Cells Newborn Infant Oxygen Pathologic Pathologic Neovascularization Pathway interactions Peripheral Phenotype Phosphotransferases Play Production Proliferating Protein Isoforms Proteins Retina Retinal Diseases Retinal Neovascularization Retinopathy of Prematurity Role Sampling Signal Pathway Signal Transduction Testing Transgenic Organisms Vascular Endothelial Growth Factors angiogenesis cell type chromatin immunoprecipitation cytokine design in vitro Model in vivo knock-down middle age mouse model neovascular neovascularization ocular angiogenesis overexpression proliferative diabetic retinopathy promoter retinal angiogenesis tool

项目摘要

PROJECT SUMMARY This proposal seeks to develop a new molecular strategy to limit pathological retinal angiogenesis in blinding eye disease. An emerging consensus is that macrophages/microglia drive aberrant neovascularization. However, the mechanisms involved remain poorly defined. Glycolysis is a major metabolic process in macrophages. PFKFB3 (6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase isoform 3, Pfkfb3 for mice) is a critical enzyme for activation of glycolysis in vascular cells and leukocytes. It catalyzes the synthesis of fructose-2, 6- bisphosphate, which is the most potent allosteric activator of 6-phosphofructo-1-kinase, a rate-limiting enzyme for glycolysis. PFKFB3 is critical for the inflammatory phenotype of macrophages, and glycolysis has been shown to promote activation of hypoxia-inducible factor (HIF) or histone acetylation signaling in some cell types. However the role of PFKFB3 in regulating angiogenesis and the underlying signaling pathways are completely unknown. Our preliminary data show that myeloid cells in the retinas of the mouse model of oxygen-induced retinopathy are hyper-glycolytic, as evidenced by high levels of glycolytic molecules and regulators/activators of glycolysis including Pfkfb3. The majority of these retinal macrophages exhibit a mixed phenotype characterized by increased levels of cell surface markers for both classical and alternative activation and increased expression of both pro-inflammatory and pro-angiogenic factors. This macrophage phenotype is recapitulated in mouse bone marrow derived macrophages treated with conditioned medium from hypoxic mouse retinal endothelial cells or the glycolytic metabolite lactate. We term these unique macrophages as Pathological Angiogenesis- associated Glycolytic Macrophages (PAGMs). Pfkfb3 knockdown in macrophages reduces their expression of pro-inflammatory and pro-angiogenic factors and suppresses neovascularization in hypoxic retinas. These data inform the hypothesis that Pfkfb3-mediated glycolysis in myeloid cells induces vascular growth by activating Hifs and histone acetylation leading to increased production of pro-inflammatory and pro-angiogenic factors. To test our hypothesis, we have generated myeloid cell specific Pfkfb3-deficient mice. We have also established methods for generating BMDMs that are Pfkfb3 deficient (hypo-glycolytic), Pfkfb3 overexpressing (hyper- glycolytic) or Pfkfb3 wild type and for knockdown of Hif1a and Hif2a. We will investigate the effect of modulating glycolysis in PAGMs on pathological angiogenesis using these genetic tools and in vivo, ex vivo, and in vitro models. We propose three specific aims. 1) Hyper-glycolytic PAGMs regulate the sprouting and proliferation of retinal or choroidal endothelial cells by production and release of pro-inflammatory and pro-angiogenic factors. 2) Hifs and histone acetylation pathways are critically involved in PAGM polarization and activation induced by endogenous glycolysis or endothelial derived glycolytic metabolites. 3) Pfkfb3-mediated glycolysis in macrophages/microglia plays an important role in the development of pathological choroidal and intraretinal neovascularization. 1

项目摘要该提案旨在制定一种新的分子策略，以限制盲目的病理视网膜血管生成眼病。新兴共识是巨噬细胞/小胶质细胞驱动异常的新生血管形成。然而，所涉及的机制的定义仍然很差。糖酵解是巨噬细胞中的主要代谢过程。 PFKFB3（6-磷酸2-激酶/果糖-2，6-二磷酸酶同工型3，PFKFB3的6-二氨基酶）是关键的酶用于激活血管细胞和白细胞中的糖酵解。它催化果糖2、6-的合成双磷酸盐是6-磷酸1-激酶的最有效的变构激活剂，一种限速酶用于糖酵解。 PFKFB3对于巨噬细胞的炎症表型至关重要，已经显示了糖酵解促进某些细胞类型中缺氧诱导因子（HIF）或组蛋白乙酰化信号的激活。但是，PFKFB3在调节血管生成和潜在信号通路中的作用完全是未知。我们的初步数据表明，氧诱导的小鼠模型的视网膜中的髓样细胞视网膜病是高糖酵解的，这是由高水平的糖酵解分子和调节剂/活化剂所证明的糖酵解包括PFKFB3。这些视网膜巨噬细胞中的大多数表现出具有特征的混合表型通过增加经典和替代激活的细胞表面标记水平和表达增加促炎和促血管生成因子。在小鼠中概括了这种巨噬细胞表型骨髓衍生的巨噬细胞用低氧小鼠视网膜内皮治疗的条件培养基处理细胞或糖酵解代谢物乳酸。我们将这些独特的巨噬细胞称为病理血管生成 - 相关的糖酵解巨噬细胞（PAGM）。巨噬细胞中的PFKFB3敲低可降低其表达促炎和促血管生成因素，并抑制缺氧视网膜中的新血管化。这些数据告知髓样细胞中PFKFB3介导的糖酵解的假设通过激活HIF诱导血管生长组蛋白乙酰化导致促炎和促血管生成因子的产生增加。测试我们的假设，我们已经产生了髓样细胞特异性PFKFB3缺陷小鼠。我们也建立了生成PFKFB3缺陷（低糖酵解），PFKFB3过表达的BMDM的方法（高 - 糖酵解）或PFKFB3野生型以及用于HIF1A和HIF2A的敲低。我们将调查调制的效果使用这些遗传工具和体内和体外，在PAGM中糖酵解病理血管生成型号。我们提出了三个具体目标。 1）高糖酵解的PAGM调节发芽和增殖视网膜或脉络膜内皮细胞通过产生和释放促炎和促血管生成因子。 2）HIF和组蛋白乙酰化途径与PAGM极化和激活有关内源性糖酵解或内皮衍生的糖酵解代谢产物。 3）PFKFB3介导的糖酵解巨噬细胞/小胶质细胞在病理脉络膜和术后发展中起重要作用新血管形成。 1

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Adenosine kinase promotes post-infarction cardiac repair by epigenetically maintaining reparative macrophage phenotype.

DOI：
10.1016/j.yjmcc.2022.11.007
发表时间：
2023-01
期刊：
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY
影响因子：
5
作者：
Zhang, Min;Wang, Caiping;Wang, Rongning;Xu, Jiean;Wang, Zhefeng;Yan, Jianlong;Cai, Yongfeng;Li, Liangping;Huo, Yuqing;Dong, Shaohong
通讯作者：
Dong, Shaohong

Gene-dosage effect of Pfkfb3 on monocyte/macrophage biology in atherosclerosis.