Role of intercellular communication in pathogenesis of diabetic retinopathy

细胞间通讯在糖尿病视网膜病变发病机制中的作用

基本信息

批准号：
8415906
负责人：
Sayon Roy
金额：
$ 34.33万
依托单位：
BOSTON MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-01-01 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8415906
关键词：
Affect Age American Apoptosis Biochemical Biological Blindness Blood Vessels Blood capillaries Cell Communication Cell Death Cell Survival Cell physiology Cells Characteristics Communication Connexin 43 Connexins Data Development Diabetes Mellitus Diabetic Retinopathy Down-Regulation Dyes Endothelial Cells Equilibrium Extracellular Matrix Extravasation Fluorescence Microscopy Functional disorder Gap Junctions Gene Expression Glucose Goals Homeostasis Hyperglycemia In Vitro Intercellular Junctions Knockout Mice Lesion Manuscripts Mediating Methods Mission Molecular Oligonucleotides Pathogenesis Pericytes Permeability Phenotype Play Production Rattus Reporting Research Retina Retinal Role Stress Techniques Testing Tight Junctions Up-Regulation Vascular Permeabilities Western Blotting Wild Type Mouse Work base capillary claudin-1 protein diabetic patient diabetic rat gap junction channel in vivo insight intercellular communication monolayer novel occludin prevent

项目摘要

The overall goal of this project is to test the hypothesis that reduced Cx43 expression and subsequent altered cell-cell communication triggered by high glucose/diabetes leads to cell death and breakdown of vascular homeostasis in the retinal capillaries in diabetic retinopathy. The hypothesis is based on findings that high glucose reduces Cx43 expression in microvascular endothelial cells and retinal pericytes and compromises gap junction intercellular communication. Our current study showed reduced Cx43 expression triggers apoptosis (manuscript under revision, IOVS). The overall working hypothesis is that reduced Cx43 expression triggers vascular cell death, a prominent and early lesion associated with the development of diabetic retinopathy, which in turn, reduces cell-cell communication in the retinal vascular cells and ultimately disrupts vascular homeostasis. Retinal vascular cell death is known to occur by apoptosis but it is unknown how apoptosis is triggered during the development of diabetic retinopathy. Preliminary data suggests that communication between vascular cells, that is endothelial cell-endothelial cell, endothelial cell-pericyte, and pericyte-pericyte is essential for their survival, and that disruption in cell-cell communication may trigger apoptosis and interfere with their role to form a functional unit via the connexin (Cx) channels in cell junctions. The specific focus of this proposal is to determine (i) the effect of reduced Cx43 expression in retinal endothelial cells and pericytes, (ii) whether high glucose- induced excess ECM synthesis modulates Cx43 expression in retinal endothelial cells and pericytes, and (iii) whether reduced Cx43 expression induces tight junction dysfunction in the retinal endothelial cells. A variety of cell biological, molecular biological and biochemical techniques including antisense oligo mediated specific downregulation of gene expression, scrape load dye transfer technique, fluorescence microscopy and Western blot method will be used for studying the consequence of high glucose-induced inhibition of Cx43 expression and reduced cell-cell communication on retinal cell viability and function. Findings from these studies will provide a better understanding of cell-cell communication underlying altered Cx43 expression and gap junction channels in retinal vascular cells and their role in breakdown of vascular homeostasis associated with diabetic retinopathy. An important mission of the agency is to find a cure and prevent complications arising from diabetic retinopathy. The proposed project is expected to provide important findings that can help in better understanding the pathogenesis of diabetic retinopathy.

该项目的总体目标是测试降低CX43表达的假设随后由高葡萄糖/糖尿病引发的细胞细胞通信发生了变化糖尿病性视网膜毛细血管中的细胞死亡和血管稳态的崩溃视网膜病。该假设是基于高葡萄糖减少CX43的发现微血管内皮细胞和视网膜周细胞中的表达并损害差距连接界面通信。我们目前的研究显示CX43降低表达会触发凋亡（根据IOV的修订，手稿）。整体工作假设是CX43表达降低会触发血管细胞死亡，这是一个突出的和与糖尿病性视网膜病的发展有关的早期病变，而糖尿病性视网膜病变又减少视网膜血管细胞中的细胞细胞通信，并最终破坏血管稳态。已知视网膜血管细胞死亡是通过凋亡发生的，但它是未知在糖尿病性视网膜病变的发展过程中如何触发凋亡。初步数据表明，血管细胞之间的通信，即内皮细胞 - 内皮细胞，内皮细胞 - 周期性细胞和周细胞周围的细胞对其生存以及细胞 - 细胞通信中的破坏可能会触发凋亡和干扰他们通过细胞中的连接素（CX）通道形成功能单元的作用连接。该提案的具体重点是确定（i）减少的影响 CX43在视网膜内皮细胞和周细胞中的表达，（ii）是否高葡萄糖诱导过多的ECM合成调节视网膜内皮细胞中的CX43表达和周细胞，以及（iii）降低的CX43表达是否诱导紧密连接视网膜内皮细胞功能障碍。各种细胞生物学，分子生物学和生化技术，包括反义寡核介导的特异性基因表达，刮擦负载染料转移技术的下调，荧光显微镜和蛋白质印迹法将用于研究高的结果葡萄糖诱导的CX43表达抑制和细胞细胞通信降低视网膜细胞活力和功能。这些研究的发现将提供更好的了解CX43表达和差距改变的细胞电池通信视网膜血管细胞中的连接通道及其在血管分解中的作用与糖尿病性视网膜病有关的稳态。该机构的重要任务是找到治愈并防止糖尿病性视网膜病变引起的并发症。提议预计项目将提供重要的发现，可以帮助更好地理解糖尿病性视网膜病的发病机理。