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

• 批准号: 8002011
• 负责人: Sayon Roy
• 金额: $ 36.14万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8002011
• 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; Health; 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

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: Currently, there is no cure for diabetic retinopathy, the leading cause of blindness in the working age Americans. It is expected that the findings from this project would provide valuable insight towards testing a novel mechanism underlying retinal vascular cell loss and capillary leakage, the two critical steps in the development of diabetic retinopathy.

描述（由申请人提供）：本项目的总体目标是检验以下假设：高糖/糖尿病引发的Cx43表达降低和随后细胞间通讯改变导致糖尿病视网膜病变中视网膜毛细血管中细胞死亡和血管稳态破坏。这一假说是基于高葡萄糖降低微血管内皮细胞和视网膜周细胞中Cx43表达并损害间隙连接细胞间通讯的发现。我们目前的研究表明，Cx43表达减少触发细胞凋亡（修订稿，IOVS）。总体工作假设是，Cx43表达减少触发血管细胞死亡，这是与糖尿病视网膜病变发展相关的突出和早期病变，这反过来又减少了视网膜血管细胞中的细胞-细胞通讯，并最终破坏血管稳态。视网膜血管细胞死亡是由细胞凋亡引起的，但在糖尿病视网膜病变的发展过程中，细胞凋亡是如何被触发的还不清楚。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-周细胞，周细胞-周细胞是必不可少的，它们的生存，细胞-细胞通信的中断可能会触发细胞凋亡，并干扰它们的作用，形成一个功能单位，通过连接蛋白（Cx）通道在细胞连接。该建议的具体重点是确定（i）视网膜内皮细胞和周细胞中Cx43表达减少的影响，（ii）高糖诱导的过量ECM合成是否调节视网膜内皮细胞和周细胞中Cx43表达，以及（iii）Cx43表达减少是否诱导视网膜内皮细胞中紧密连接功能障碍。本研究将采用多种细胞生物学、分子生物学和生物化学技术，包括反义寡核苷酸介导的特异性基因表达下调、刮载染料转移技术、荧光显微镜和Western blot方法，研究高糖诱导的Cx43表达抑制和细胞间通讯减少对视网膜细胞活力和功能的影响。这些研究的结果将提供一个更好的了解视网膜血管细胞中Cx43表达和间隙连接通道改变的细胞间通讯及其在糖尿病视网膜病变相关血管稳态破坏中的作用。该机构的一项重要使命是找到治疗方法并预防糖尿病视网膜病变引起的并发症。该项目有望提供重要的发现，有助于更好地了解糖尿病视网膜病变的发病机制。公共卫生相关性：目前，糖尿病视网膜病变无法治愈，而糖尿病视网膜病变是美国工作年龄段失明的主要原因。预计该项目的发现将为测试视网膜血管细胞损失和毛细血管渗漏的新机制提供有价值的见解，这是糖尿病视网膜病变发展的两个关键步骤。