The Role of the Nicotinic Cholinergic Pathway in Retinopathy of Prematurity

烟碱胆碱能通路在早产儿视网膜病变中的作用

基本信息

批准号：
8334482
负责人：
JOHN P COOKE
金额：
$ 40.52万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8334482
关键词：
Acetylcholine Affect Angiogenic Factor Arterial Fatty Streak Biochemical Biological Birth Blindness Blood Vessels Child Choline O-Acetyltransferase Cholinergic Receptors Choroid Choroidal Neovascularization Development Disease Drug Formulations Elements Endothelial Cells Exudative age-related macular degeneration Eye Genetic Growth Factor Human Volunteers Hypoxia Immunohistochemistry Impairment In Situ Hybridization Infant Inflammatory Ischemia Knockout Mice Lead Learning Ligands Malignant neoplasm of lung Mecamylamine Mediating Medicine Methods Modeling Mus Nature Neurons Nicotinic Receptors Pathogenesis Pathologic Neovascularization Pathway interactions Patients Permeability Phase Phase II Clinical Trials Phenotype Plasma Pregnancy Premature Infant Receptor Activation Research Retina Retinal Retinal Detachment Retinal Diseases Retinal Neovascularization Retinopathy of Prematurity Reverse Transcriptase Polymerase Chain Reaction Risk Factors Role Safety Sclera Signal Transduction Signaling Molecule Stimulation of Cell Proliferation Time Traction Tube Tumor Angiogenesis United States Vascular Endothelial Growth Factors Vascularization Vision angiogenesis base cancer cell choline transporter cholinergic diabetic human subject insight laser capture microdissection macular edema methyllycaconitine migration neovascularization neuron development novel novel strategies novel therapeutic intervention novel therapeutics phase 1 study prevent receptor retina blood vessel structure retinal neuron tumor growth

项目摘要

Retinopathy of prematurity (ROP), is a leading cause of vision impairment and blindness in children in the United States, due to pathological retinal neovascularization. We have discovered a novel angiogenic pathway that is involved in pathological neovascularization. This pathway is mediated by an endothelial nicotinic acetylcholine receptor (nAChR). The endothelial nAChR is a ligand-gated cationic channel that is activated by the endogenous signaling molecule, acetylcholine (ACh). Activation of this receptor induces endothelial cell mitogenesis, migration and tube formation and promotes angiogenesis. The pathway is upregulated by hypoxia, and by other angiogenic factors such as the vascular endothelial growth factor (VEGF). Accordingly, we propose the following Specific Aims to develop a novel therapy for ROP: 1. Characterize the endothelial nicotinic cholinergic pathway in the developing retina, and determine its role in normal vascularization. We hypothesize that normal development of the retina will not be adversely affected by pharmacological antagonism of the EC nAChRs. This hypothesis is based in part on the normal phenotype of the ¿7-nAChR deficient mouse. We will begin by determining, during normal development, the level of expression and localization of key elements of the nAChR pathway including the high affinity choline transporter, choline acetyltransferase, and the nAChR subunits. We will study EC from normal retinal vessels using laser capture microdissection and real time RT-PCR, in situ hybridization and immunohistochemistry. We will carefully assess vascular and neuronal development in the retina of the ¿7-nAChR deficient mouse. Finally, we will determine if pharmacological antagonism of nAChRs in the eye (comparing non-selective versus ¿7-nAChR selective antagonists) will adversely effect normal neuronal or vascular development. 2. Determine if excessive activation of this pathway contributes to retinal neovascularization in a murine model of ROP, using pharmacological and genetic knockdown of EC nAChRs. Using the methods described above, we will determine if retinal neovascularization is associated with increased retinal expression of any components of the nAChR pathway. We will determine if administration of the non-selective (mecamylamine) or ¿7-preferential (methyllycaconitine) nAChR antagonists suppress retinal neovascularization. The effect of nAChR antagonists on retinal neuronal function will be assessed by ERGs. ROP will be induced in ¿7- nAChR knockout mice and littermate controls to determine if signaling through ¿7- nAChRs contributes to retinal neovascularization. We will perform biochemical studies to assess the relationship between plasma and retinal levels of ACh, and correlate these levels to changes in retinal levels of vascular endothelial growth factor (VEGF), retinal vascularity and permeability.

早产视网膜病（ROP）是视力障碍和失明的主要原因由于病理视网膜新血管形成，美国。我们发现了一种新型的血管生成途径这与病理新生血管形成有关。该途径是由内皮烟碱介导的乙酰胆碱受体（NACHR）。内皮NACHR是一个配体门控阳离子通道，被激活内源信号分子乙酰胆碱（ACH）。该受体的激活诱导内皮细胞有丝分裂，迁移和管形成并促进血管生成。该路径已更新缺氧，以及其他血管生成因子，例如血管内皮生长因子（VEGF）。我们提出了以下特定目的，以开发一种新的ROP疗法： 1。表征发育中的视网膜中的内皮烟碱胆碱能途径，并确定其在正常血管中的作用。我们假设视网膜的正常发展不会对受EC NACHR的药物拮抗作用的影响。该假设部分基于正常 7-NACHR缺乏小鼠的表型。我们将首先确定在正常发展期间 NACHR途径的关键要素的表达水平和定位水平包括高亲和力胆碱转运蛋白，胆碱乙酰转移酶和NACHR亚基。我们将从正常残留血管中研究EC 使用激光捕获显微解剖和实时RT-PCR，原位杂交和免疫组织化学。我们将仔细评估7-NACHR缺乏小鼠视网膜中的血管和神经元发育。最后，我们将确定眼睛中NACHR的药物拮抗作用（比较非选择性）与7-NACHR选择性拮抗剂）将对正常的神经元或血管发育产生不利影响。 2。确定该途径过度激活是否有助于视网膜新血管化 ROP的鼠模型，使用EC NACHR的药物和遗传敲低。使用上述方法，我们将确定残留的新血管形成是否与视网膜增加有关 NACHR途径的任何组件的表达。我们将确定非选择性的给药（美甲基胺）或7首发（甲基粘液氨酸）NACHR拮抗剂抑制残留新血管形成。 NACHR拮抗剂对永久神经元功能的影响将由ERG评估。 ROP将在7-NACHR敲除小鼠和同窝立物中诱导，以确定是否通过7-信号传导。 NACHR有助于视网膜新血管形成。我们将进行生化研究以评估血浆与ACH的残留水平之间的关系，并将这些水平与永久水平的变化相关联血管内皮生长因子（VEGF），视网膜血管和渗透性。