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（由适用提供）：糖尿病性视网膜病（DR）是发达国家视力丧失的主要原因，影响了1型糖尿病患者（T1DM）的90％，至少15年疾病持续时间。诸如激光光凝和抗VEGF药物之类的治疗通常能够治疗增殖的糖尿病性视网膜病（PDR）和黄斑水肿；但是，基于归因于高血糖的机制的早期阶段的临床治疗尚未成功。由于流行病学观察结果是，乔斯林50年的奖章研究队列（N> 900）是一组具有50年或更长时间的T1DM的组，并未发展出严重的DR（35％）所需的有限早期的血糖控制工具，以实现血糖控制的早期工具，我们专注于寻找一个内源性视网膜保护因子。使用视网膜和玻璃体的蛋白质组学分析，与患有PDR的奖章者相比，即使在高血糖症的情况下，也发现了那些具有MILD DR的奖章者中的视黄蛋白结合蛋白3（RBP3）升高。初步数据表明，与主动或静止的PDR相比，T1D和T2D患者的这种玻璃体中的RBP3升高模式是一致的。纯化的RBP3抑制了高血糖和VEGF对残留内皮细胞和周细胞的作用。有趣的是，纯化的RBP3刺激了纳摩尔浓度的周细胞和内皮细胞中P-AKT和P-ERK的信号级联。通过视网膜下注射慢病毒的过度表达可阻止毛细血管渗透性和VEGF表达的增加，通过ERG和OCT测量的神经视网膜功能障碍，在糖尿病性Lewis大鼠中形成了无质的毛细血管和周细胞凋亡。这些研究表明，RBP3除了在感光细胞和视网膜猪上皮细胞之间转运性类动物以外的功能。为了确认RBP3可以保护视网膜中的新作用，以防止高血糖的不利影响，我们提出了三个具体目的：特定目的1：评估非糖尿病控制中的玻璃体和水性液体中的RBP3水平，从非糖尿病控制中，患有1型和类型1型和2型糖尿病患者的各种持续时间，年龄，年龄，严重的DR和GLECLECECTOCMECTOCMECTIC COTTROT。具体目标2：确定在视网膜中升高RBP3水平还是玻璃体是否可以降低啮齿动物中糖尿病引起的视网膜或血管异常。特定目的3：表征RBP3的信号传导和视网膜周围和内皮细胞中的作用。