Identification of Retinoid-Binding Protein 3 (RBP3): A Protective Factor against Diabetic Retinopathy Using Retina from People with Extreme Duration of Diabetes

类维生素A结合蛋白3 (RBP3)的鉴定：利用糖尿病病程极长的人的视网膜来鉴定糖尿病视网膜病变的保护因子

• 批准号: 9006846
• 负责人: GEORGE L KING
• 金额: $ 41.38万
• 依托单位: JOSLIN DIABETES CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9006846
• 关键词: Adverse effects; Affect; Age; Antibodies; Apoptosis; Background Diabetic Retinopathy; Binding Proteins; Biological Assay; Biological Markers; Blindness; Blood Vessels; Blood capillaries; Capillary Permeability; Cattle; Cells; Chronic; Clinical Treatment; Clinical Trials; Cohort Studies; Data; Developed Countries; Diabetes Mellitus; Diabetic Retinopathy; Disease; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Epidemiology; Exhibits; Eye; Functional disorder; Glucose; Glycosylated hemoglobin A; Hyperglycemia; Individual; Injection of therapeutic agent; Insulin-Dependent Diabetes Mellitus; Intervention; Intervention Studies; Liquid substance; Mass Spectrum Analysis; Measures; Microvascular Dysfunction; Neural Retina; Non-Insulin-Dependent Diabetes Mellitus; Patients; Pattern; Pericytes; Permeability; Photoreceptors; Prevalence; Proteins; Proteomics; Proto-Oncogene Proteins c-akt; Rattus; Retina; Retinal; Retinal Photoreceptors; Retinoids; Retinol Binding Proteins; Rhodopsin; Rodent; Severities; Signal Transduction; Site; Staging; Structure; Structure of retinal pigment epithelium; Subfamily lentivirinae; Testing; Therapeutic; Therapeutic Intervention; Thick; Tissues; Toxic effect; Transgenic Mice; Vascular Endothelial Growth Factors; aqueous; base; capillary; cohort; diabetic; diabetic patient; glycemic control; illness length; in vivo; interstitial retinol-binding protein; laser photocoagulation; macular edema; migration; nanomolar; non-diabetic; novel; overexpression; prevent; proliferative diabetic retinopathy; promoter; public health relevance; receptor; relating to nervous system; retina blood vessel structure; retinal apoptosis; subretinal injection; tool; type I and type II diabetes; type I diabetic; vascular abnormality

 DESCRIPTION (provided by applicant): Diabetic retinopathy (DR) is the leading cause of vision loss in developed countries and affects 90% of type 1 diabetic patients (T1DM) with at least 15 years disease duration. Treatments such as laser photocoagulation and anti-VEGF agents are often able to treat proliferative diabetic retinopathy (PDR) and macular edema; however, clinical treatment for early stages of DR based on the mechanisms attributed to hyperglycemia have not been successful. Due to the epidemiological observation that the Joslin 50-Year Medalist Study cohort (n>900), a group with 50 or more years of T1DM, did not develop severe DR (prevalence 35%) despite limited early tools for glycemic control, we focused on finding an endogenous retinal protective factor. Using proteomic analysis of the retina and vitreous, retinoid binding protein-3 (RBP3) was found to be elevated in those Medalists with no-mild DR compared to those with PDR even in the presence of hyperglycemia. Preliminary data showed that this pattern of RBP3 elevation in the vitreous with no-mild DR compared to active or quiescent PDR is consistent in both T1D and T2D patients. Purified RBP3 inhibited the actions of hyperglycemia and VEGF on retinal endothelial cells and pericytes. Interestingly, purified RBP3 stimulated signaling cascades of p-AKT and p-ErK in pericytes and endothelial cells at nanomolar concentrations. Overexpression of RBP3 by subretinal injections of lentivirus prevented increases in capillary permeability and VEGF expression, neural retinal dysfunction measured by ERG and OCT, the formation of acellular capillaries and pericyte apoptosis in diabetic Lewis rats, all suggest the protective ability of the RBP3 protein. These studies suggest that RBP3 has functions other than transporting retinoids between photoreceptors and retinal pigmented epithelial cells. To confirm that RBP3 could have protective and novel actions in the retina against the adverse effects of hyperglycemia, we propose three specific aims: Specific Aim 1: To evaluate RBP3 levels in the vitreous and aqueous fluids from non-diabetic controls, people with type 1 and type 2 diabetes of various duration, age, severity of DR and glycemic control. Specific Aim 2: To determine whether elevating the level of RBP3 in the retina or vitreous can decrease retinal or vascular abnormalities induced by diabetes in rodents. Specific Aim 3: To characterize RBP3's signaling and actions in retinal pericyte and endothelial cells.

 描述（由申请人提供）：糖尿病视网膜病变 (DR) 是发达国家视力丧失的主要原因，影响 90% 的 1 型糖尿病患者 (T1DM)，病程至少为 15 年。 VEGF 药物通常能够治疗增殖性糖尿病视网膜病变 (PDR) 和黄斑水肿；然而，早期 DR 的临床治疗基于以下机制：由于流行病学观察，Joslin 50 年奖章获得者研究队列 (n>900)（一组患有 50 年或以上 T1DM）并未发展为严重 DR（患病率 35%），尽管早期工具有限。为了控制血糖，我们重点寻找一种内源性视网膜保护因子，通过对视网膜和玻璃体的类维生素A结合蛋白3进行蛋白质组学分析。研究发现，即使在存在高血糖的情况下，非轻度 DR 的获奖者的 RBP3 水平也比 PDR 的获奖者升高。 PDR 在 T1D 和 T2D 患者中是一致的，纯化的 RBP3 抑制高血糖和 VEGF 对视网膜内皮细胞的作用。在周细胞中，纯化的 RBP3 在纳摩尔浓度下刺激周细胞和内皮细胞中的 p-AKT 和 p-ErK 信号级联反应，通过视网膜下注射慢病毒来防止毛细血管通透性和 VEGF 表达的增加，以及通过 ERG 和OCT、糖尿病Lewis大鼠脱细胞毛细血管的形成和周细胞凋亡都表明了这些研究表明，RBP3 除了在光感受器和视网膜色素上皮细胞之间运输类视黄醇外，还具有其他功能。为了证实 RBP3 可以在视网膜中对抗高血糖的不利影响具有保护性和新颖的作用，我们提出了三个具体目标：目标 1：评估非糖尿病对照者、各种 1 型和 2 型糖尿病患者的玻璃体和房水中的 RBP3 水平具体目标 2：确定提高视网膜或玻璃体中的 RBP3 水平是否可以减少啮齿类动物糖尿病引起的视网膜或血管异常。存在于视网膜周细胞和内皮细胞中。