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: 8332428
• 负责人: Sayon Roy
• 金额: $ 5.55万
• 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/8332428
• 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）：该项目的总体目标是测试以下假设：高葡萄糖/糖尿病触发的CX43表达和随后改变的细胞 - 细胞通信导致细胞死亡和糖尿病性视网膜病变视网膜毛细管中血管稳态的细胞死亡。该假设是基于发现高葡萄糖会降低微血管内皮细胞和视网膜周细胞中CX43表达的发现，并损害间隙连接间的细胞间通信。我们目前的研究表明，CX43表达降低会触发凋亡（根据IOV的手稿）。总体工作假设是，降低的CX43表达会触发与糖尿病性视网膜病的发展有关的血管细胞死亡，这是一种突出的和早期的病变，进而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细胞凋亡发生的，但尚不清楚糖尿病性视网膜病变过程中凋亡是如何触发的。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 - 细胞 - 周期性细胞和周细胞周期对生存至关重要，细胞细胞通信的破坏可能会触发凋亡和干扰其通过CONNEXIN（CX）CYLNEXIN（CX）Channels in narneels in narneels in narneels in narneels in narcintels in narnections in narcintions in a connexin。 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.各种细胞生物学，分子生物学和生物化学技术，包括反义寡聚介导的基因表达的特异性下调，Scrape载荷染料转移技术，荧光显微镜和蛋白质印迹方法将用于研究高葡萄糖诱导的CX43表达抑制和降低细胞电池对维持细胞的抑制作用，从而研究导致维持细胞的抑制作用。这些研究的发现将更好地了解视网膜血管细胞中CX43表达和间隙连接通道的基础细胞通信，以及它们在与糖尿病性视网膜病变相关的血管稳态分解中的作用。该机构的一个重要任务是找到治疗方法并防止因糖尿病性视网膜病变引起的并发症。预计该项目将提供重要的发现，可以更好地理解糖尿病性视网膜病的发病机理。公共卫生相关性：目前，无法治愈糖尿病性视网膜病，这是美国人劳动时代失明的主要原因。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泄漏的新机制提供宝贵的见解，这是糖尿病性视网膜病变发展的两个关键步骤。