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：
8473864
负责人：
Maria Bartolomeo Grant
金额：
$ 36.58万
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 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8473864
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 endothelial dysfunction 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 public health relevance 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. PUBLIC HEALTH RELEVANCE: Endothelial precursors cells (EPCs) are bone marrow derived cells that have been used as novel therapies for ischemic conditions currently lacking effective treatment options. The identification, manipulation and transplantation of these cells are key to optimizing their use; however, diabetic patients have defective EPCs that cannot be used for cellular therapy such for repair of the vasodegenerative phase (acellular capillaries) of diabetic retinopathy. The focus of this application is to 1) identify the optimal EPC populations for repair, 2) characterize the defects in diabetic EPCs, 3) repair the defects, 4) optimize the timing and dosage of the EPC injections and identify the changes in the diabetic bone marrow microenvironment. These studies will be performed using human EPCs from diabetics with varying degrees of retinopathy, animal models of diabetes and knockout mice with unique characteristics that impact the bone marrow microenvironment.

描述（由申请人提供）：内皮祖细胞（EPC）功能障碍可能在DR的发病机理中具有关键作用。由培养的单核细胞（MNC）引起的两种EPC人群是晚产生的内皮细胞（OEC），它们显示出克隆表型，属于CD34+CD45-种群；和早期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。为了解决这些细胞在糖尿病性视网膜病（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，在非增殖性糖尿病性视网膜病（NPDR）中，OEC人群瞬时损失，但它再次出现为更具侵略性和增生性的人群，从而触发了“血管生成”转换和增殖的发作（PDR）。相比之下，EEPC人群从未完全消失，而是在其活动水平上转移，PDR的炎症性更大，NPDR较少。在这两个种群中，一氧化氮合酶（NOS）的失调对于这些表型转变都是核心，这些跃迁进一步受到骨髓（BM）微环境的变化影响。我们正在通过以下目的检验这一假设：具体目的1：测试EEPC和OEC的增殖潜力取决于视网膜病的阶段，并确定EEPC和OEC是否从糖尿病个体中分离出来，是否产生更具反应性的氧物种产生更具反应性氧的物种，并包含更具内源性的Nos抑制剂，并含有更大的细胞和分泌的细胞。具体目的2：测试非糖尿病（健康）EEPC和OEC的组合是在视网膜缺血 - 再灌注损伤模型中单独的人群或糖尿病SCID小鼠中的细胞毛细血管中单独的人群的组合。特定目标3：测试糖尿病中的EEPC和OEC功能障碍是否是由于BM微环境中的缺陷（肥胖增加，造血减少和祖细胞功能障碍）引起的。这些研究将使我们能够确定EEPC和OEC是否代表了细胞疗法的理想祖细胞种群，以改善糖尿病眼中血管的健康。公共卫生相关性：内皮前体细胞（EPC）是骨髓衍生的细胞，这些细胞已被用作目前缺乏有效治疗选择的缺血性疾病的新疗法。这些细胞的识别，操纵和移植是优化它们使用的关键。然而，糖尿病患者的EPC有缺陷，无法用于细胞治疗，例如用于修复糖尿病性视网膜病的血管降节期（毛细血管毛细血管）。该应用的重点是1）确定最佳的EPC人群以进行修复，2）表征糖尿病EPC中的缺陷，3）修复缺陷，4）优化EPC注射的时间和剂量，并确定糖尿病骨髓微环境的变化。这些研究将使用来自具有不同程度的视网膜病变，糖尿病动物模型和敲除小鼠的人类EPC进行，具有影响骨髓微环境的独特特征。