Dysfunction of endothelial precursor subtypes dictates the outcomes of diabetic r

内皮前体亚型的功能障碍决定了糖尿病患者的结局

基本信息

批准号：
8730783
负责人：
Maria Bartolomeo Grant
金额：
$ 36.8万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8730783
关键词：
Actins Address Adipocytes Agreement Angiogenic Switch Angiopoietin-1 Animal Model Animals Background Diabetic Retinopathy Behavior Bioavailable Biological Assay Biological Markers Blood Vessels Blood capillaries Bone Marrow Bone Marrow Transplantation Bromodeoxyuridine CD14 gene CD34 gene CXCL12 gene Cell Death Cell Transplantation Cells Characteristics Cyclic GMP Defect Development Diabetes Mellitus Diabetic Retinopathy Diabetic mouse Endothelial Cells Environment Enzyme-Linked Immunosorbent Assay Epoprostenol Equilibrium Ethers Exhibits Eye Financial compensation Fluorescence Functional disorder Funding Generations Glucose Health Hematopoiesis Histology Human Hydrolase Hypoxia IL8 gene In Vitro Individual Inflammatory Injection of therapeutic agent Insulin-Like Growth Factor I Interleukin-1 Investigation Knockout Mice Lead Magnetism Measures Metalloproteases Modeling Molecular Mononuclear Motor Mus N,N-dimethylarginine NADPH Oxidase Nature Neurons Nitric Oxide Nitric Oxide Donors Nitric Oxide Synthase Obesity Outcome Oxidation-Reduction PTPRC gene Pathogenesis Pathologic Neovascularization Patients Peroxisome Proliferator-Activated Receptors Phase Phenotype Phosphorylation Physiological Population Process Proteins Reactive Oxygen Species Reperfusion Injury Retina Retinal Retinal Diseases Rodent Role SCID Mice Signal Transduction Site Sorting - Cell Movement Staging Stem cells Streptozocin Surface TNF gene Testing Therapeutic Time Transplantation Tube Vascular Endothelial Growth Factor Receptor-2 Vascular Endothelial Growth Factors Wild Type Mouse angiogenesis base bisphenol A capillary cell motility cytokine diabetic diabetic patient dosage effective therapy enzyme activity human NOS3 protein improved in vitro Assay in vivo inhibitor/antagonist lipid biosynthesis migration monocyte neovascularization non-diabetic novel peripheral blood precursor cell progenitor reconstitution repaired research study response retinal ischemia translational study vascular bed vasodilator-stimulated phosphoprotein

项目摘要

DESCRIPTION (provided by applicant): Endothelial progenitor cell (EPC) dysfunction may have a key role in the pathogenesis of DR. Two populations of EPCs that arise from cultured mononuclear cells (MNC) are the late outgrowth endothelial cells (OECs), which display a clonal phenotype and belong to a CD34+CD45- population; and early EPCs (eEPCs) which exhibit a monocyte phenotype (CD14+/CD45+) but demonstrate endothelial-like markers and behavior in vitro and in vivo. To address the role of these cells in diabetic retinopathy (DR) we hypothesized that in DR, eEPCs remain as the central modulator of the OECs; however, the OEC population is transiently lost in nonproliferative diabetic retinopathy (NPDR) but it reappears as a more aggressive and proliferative population which triggers the "angiogenic" switch and the onset of proliferative (PDR). The eEPC population, in contrast, never disappears entirely but rather shifts in its level of activity, being more inflammatory in PDR and less in NPDR. In both populations, dysregulation of nitric oxide synthase (NOS) is central to these phenotypic transitions, which are further influenced by the changing bone marrow (BM) microenvironment. We are testing this hypothesis by the following aims: Specific Aim 1: To test whether the proliferative potential of eEPCs and OECs depends on the stage of retinopathy and determine whether eEPCs and OECs isolated from diabetic individuals produce more reactive oxygen species, contain more endogenous NOS inhibitors, and secrete a distinct cytokine profile than cells from controls. Specific Aim 2: To test whether the combination of nondiabetic (healthy) eEPCs and OECs will have a greater reparative function then either population alone in models of retinal ischemia-reperfusion injury or in acellular capillaries in diabetic SCID mice. Specific Aim 3: To test whether eEPC and OEC dysfunction in diabetes is due to defects (increased adiposity, reduced hematopoiesis and progenitor dysfunction) in the BM microenvironment. These studies will allow us to determine whether eEPCs and OECs represent ideal progenitor populations for cellular therapy to improve the health of the vasculature in the diabetic eye.

描述（由申请人提供）：内皮祖细胞（EPC）功能障碍可能在 DR 的发病机制中发挥关键作用。由培养的单核细胞 (MNC) 产生的两个 EPC 群体是晚期生长内皮细胞 (OEC)，它们表现出克隆表型并属于 CD34+CD45- 群体；早期 EPC (eEPC) 表现出单核细胞表型 (CD14+/CD45+)，但在体外和体内表现出内皮样标记和行为。为了阐明这些细胞在糖尿病视网膜病变 (DR) 中的作用，我们假设在 DR 中，eEPC 仍然是 OEC 的中央调节剂；然而，在非增殖性糖尿病视网膜病变 (NPDR) 中，OEC 群体会暂时消失，但它会以更具攻击性和增殖性的群体形式重新出现，从而触发“血管生成”开关和增殖 (PDR) 的开始。相比之下，eEPC 群体从未完全消失，而是其活动水平发生了变化，在 PDR 中炎症性更强，在 NPDR 中炎症性更少。在这两个人群中，一氧化氮合酶（NOS）的失调是这些表型转变的核心，并进一步受到骨髓（BM）微环境变化的影响。我们正在通过以下目标检验这一假设：具体目标 1：测试 eEPC 和 OEC 的增殖潜力是否取决于视网膜病变的阶段，并确定从糖尿病个体中分离的 eEPC 和 OEC 是否会产生更多的活性氧、含有更多的内源性 NOS抑制剂，并分泌与对照细胞不同的细胞因子谱。具体目标 2：测试非糖尿病（健康）eEPC 和 OEC 的组合在视网膜缺血再灌注损伤模型中或糖尿病 SCID 小鼠的无细胞毛细血管中是否比单独群体具有更大的修复功能。具体目标 3：测试糖尿病中的 eEPC 和 OEC 功能障碍是否是由于 BM 微环境缺陷（肥胖增加、造血功能减少和祖细胞功能障碍）所致。这些研究将使我们能够确定 eEPC 和 OEC 是否代表细胞治疗的理想祖细胞群，以改善糖尿病眼部脉管系统的健康。