Vasculogenesis and hyperoxia in the developing retina.

视网膜发育中的血管发生和高氧。

• 批准号: 8250344
• 负责人: Gerard Anthony Lutty
• 金额: $ 47.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-12-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8250344
• 关键词: 3-Dimensional; Address; Angioblast; Astrocytes; Attenuated; Autologous; Axon; Blood Cells; Blood Vessels; Blood capillaries; CXCR4 gene; Canis familiaris; Cell Nucleus; Cell Proliferation; Cells; Coculture Techniques; Cues; Cytokine Signaling; Development; Diabetic Retinopathy; Embryo; Endothelial Cells; Engraftment; Extracellular Matrix; Eye; Fibroblast Growth Factor 2; Future; Gel; Goals; Homing; Human; Human Development; Hyperoxia; Immunohistochemistry; In Vitro; Individual; Insulin-Like Growth Factor I; Intravenous; Knowledge; LIF gene; Literature; Macrophage Colony-Stimulating Factor; Magnetism; Manuscripts; Microglia; Modeling; Muller' s cell; Mus; Natural regeneration; Neonatal; Nerve Fibers; Neurons; Neuropilin-1; Outcome; Oxygen; PGF gene; Pattern; Pericytes; Persons; Process; Regenerative Medicine; Retina; Retinal; Retinal Diseases; Retinal Neovascularization; Retinopathy of Prematurity; Severities; Signal Transduction; Source; Staging; Stimulus; Stromal Cell-Derived Factor 1; Therapeutic; Therapeutic Intervention; Therapeutic Uses; Time; Tube; Umbilical Cord Blood; VEGF165; Vascularization; Vision; angiogenesis; capillary; cell type; clinically relevant; cytokine; fetal; ganglion cell; in vivo; induced pluripotent stem cell; injured; magnetic field; migration; nanofiber; nanoparticle; neovascularization; platelet-derived growth factor BB; prevent; progenitor; public health relevance; regenerative therapy; retina blood vessel structure; vasculogenesis

DESCRIPTION (provided by applicant): Development of the human retinal vasculature is poorly understood. In the past five years, we have contributed to our understanding of this process by demonstrating that the human superficial retinal vasculature forms by vasculogenesis. Vasculogenesis entails the differentiation and assembly of vascular progenitors, angioblasts, into primordial blood vessels. We have cultured human retinal angioblasts (CXCR4+, cKit+, CD39+) from avascular neonatal dog and fetal human retina. Our overall hypothesis is that cytokines cause differentiation of these retinal angioblasts into endothelial cells (EC) and pericytes and regulate their assembly via vasculogenesis to form the superficial retinal vasculature. Furthermore, we propose that Muller and/or ganglion cells (GC) provide the guidance for this assembly. One goal of these studies is to characterize human retinal angioblasts in vitro and investigate the potential of these human cells to differentiate into various cell types found in inner retina. We will also determine the stimulus for their formation into blood vessels in vitro. We will determine the effects of VEGF165 and VEGF165b on retinal angioblast differentiation and tube formation in vitro and determine why VEGF165b is associated with nucleus of progenitors while VEGF165 is cytoplasmic and diffusely in the perivascular milieu. We will determine the cell types and extracellular matrix components that potentially provide guidance for assembly of retinal angioblasts into the fetal human retinal vasculature in vivo by immunohistochemistry and in vitro using GC and Muller cells co-cultured with angioblasts in 3-D gels. Coated nanofibers will be used to examine the influence of individual matrix components and guidance molecules (neuropilin-1, astrocytes 3A) on angioblast alignment, patterning, and assembly of tubes. These studies will broaden our knowledge of human vascular development and characterize the pertinent progenitors. It appears from the literature that the mouse superficial vasculature develops by angiogenesis and guidance for vascular assembly is provided by an astrocytic template. There is no strocytes template in advance of the superficial human retinal vasculature while developing by vasculogenesis. Mice deficient in superficial retinal blood vessels, GC or Muller cells will be used to demonstrate functionally the importance of these cells to retinal vascular development with the ultimate outcome being delineation of which features in mouse retinal vascular development can actually be transcribed to human retinal vascular development. Finally, the potential for therapeutic use of angioblasts in the initial stage of OIR will also be determined using human angioblasts from different sources (including clinically relevant iPS from cord blood cells) and magnetic nanoparticles to increase their homing and engraftment to obliterated retinal vascular segments. If acellular vascular segments can be quickly repopulated and the blood vessels regenerated, then vaso- obliteration will be limited and subsequent neovascularization could be suppressed. PUBLIC HEALTH RELEVANCE: This study will broaden our knowledge of human vascular development by characterizing the pertinent progenitors (angioblasts) and determining the cues needed to assemble the human superficial retinal vasculature. The potential of human angioblasts derived from several sources (including induced-pluripotent cells from cord blood) to be used for regenerative medicine (repopulating acellular vascular segments) will be determined using the mouse oxygen-induced retinopathy model for retinopathy of prematurity (ROP). If we want to effectively treat ROP, which is an oxidative insult of a developing vasculature, we must understand how the human retinal vasculature forms and investigate the potential for using autologous cells (angioblasts from the persons own cord blood) as a treatment.

描述（由申请人提供）：人类视网膜脉管系统的发展知之甚少。在过去的五年中，我们通过证明人类浅表性视网膜血管形成血管发生来为我们的理解做出了贡献。血管生成需要使血管祖细胞（血管细胞）分化为原始血管。我们已经从血管新生儿狗和胎儿人视网膜中培养了人类视网膜血管细胞（CXCR4+，CKIT+，CD39+）。我们的总体假设是，细胞因子会导致这些视网膜血管细胞分化为内皮细胞（EC）和周细胞，并通过血管生成调节其组装，形成浅表性视网膜脉管系统。此外，我们建议Muller和/或神经节细胞（GC）为该组件提供指导。 这些研究的目标之一是在体外表征人类视网膜血管细胞，并研究这些人类细胞分化为内部视网膜中各种细胞类型的潜力。我们还将确定它们在体外形成血管的刺激。我们将在体外确定VEGF165和VEGF165B对视网膜血管细胞分化和管形成的影响，并确定为什么VEGF165B与祖细胞核有关，而VEGF165在血管性乳房中是细胞质且弥漫性的。我们将确定细胞类型和细胞外基质成分有可能通过免疫组织化学和使用GC和Muller细胞在3-D凝胶中与血管细胞共培养的GC和Muller细胞在体内和体外在体内的体内和体外进行体内的视网膜血管细胞的指导。涂层的纳米纤维将用于检查单个基质组件和引导分子（Neuropilin-1，星形胶质细胞3A）对管胞细胞比对，模式和管子组装的影响。这些研究将扩大我们对人类血管发展的了解，并表征相关的祖细胞。 从文献看来，小鼠浅表血管通过血管生成和血管组件的指导而形成，由星形胶质细胞模板提供。在通过血管生成发展的同时，在浅表人类视网膜脉管系统之前，没有strocytes模板。缺乏浅表性视网膜血管，GC或Muller细胞缺乏的小鼠在功能上将这些细胞在视网膜血管发育中的重要性，最终结果是描述哪些在小鼠视网膜血管发育中的特征实际上可以转录为人视网膜血管发育。 最后，还将使用来自不同来源的人类血管细胞（包括来自脐带血细胞的临床相关的IP）和磁性纳米粒子来确定在OIR初始阶段使用血管细胞的治疗潜力，以增加其归巢和雕刻到闭塞视网膜血管段。如果可以快速重新填充细胞细胞血管段并再生血管，则血管抑制将受到限制，随后的新血管形成可以抑制。 公共卫生相关性：这项研究将通过表征相关的祖细胞（血管生成）并确定组装人类浅表性视网膜血管所需的线索来扩大我们对人类血管发育的了解。使用小鼠氧气诱导的视网膜病变模型来确定，从多个来源（包括脐带血的诱导性细胞）得出的人类血管细胞的潜力将用于再生医学（重新填充的细胞血管段）。如果我们想有效治疗ROP，这是对发育中的脉管系统的氧化侮辱，我们必须了解人类视网膜脉管系统如何形成并研究使用自体细胞（来自人自己的脐带血的血管细胞）作为治疗的潜力。