Neutrophil elastase and Gasdermin D in diabetic retinopathy

中性粒细胞弹性蛋白酶和 Gasdermin D 在糖尿病视网膜病变中的作用

基本信息

批准号：
10279365
负责人：
Timothy S Kern
金额：
$ 39.25万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-30 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10279365
关键词：
Acute Lung Injury Adherence Animals Area Binding Blood Vessels Blood capillaries CASP1 gene Cell Death Cell Survival Cell membrane Cells Cessation of life Characteristics Cleaved cell Cytoplasmic Granules Data Development Diabetes Mellitus Diabetic Retinopathy Diabetic mouse Elastases Endothelial Cells Endothelium Eyedrops Failure Growth Individual Inflammation Inflammatory Intercellular adhesion molecule 1 Laboratories Lead Leukocyte Elastase Leukocytes Mediating Membrane Mus Pathogenesis Pathway interactions Patients Peptide Hydrolases Pharmacology Play Process Proteins Research Research Proposals Research Subjects Retina Retinal Degeneration Retinal Diseases Retinal Neovascularization Role Secondary to Serine Protease Signal Transduction Structure Systemic Therapy Toxic effect Vascular Endothelial Growth Factors cell injury cytotoxic cytotoxicity diabetic effective therapy extracellular vesicles in vivo insight neutrophil neutrophil elastase inhibitor non-diabetic novel novel therapeutics pancreatic elastase II pathogenic microbe response to injury retinal ischemia therapeutic target tissue injury

项目摘要

PROJECT SUMMARY Considerable data now suggests that inflammatory processes play a critical role in the pathogenesis of diabetic retinopathy. Leukocytes in particular appear to play a major role in the diabetes-induced degeneration of retinal capillaries (which sets up the conditions for eventual development of retinal ischemia, release of vaso- proliferative factors like VEGF, and ultimately, retinal neovascularization). How leukocytes mediate this capillary cell damage in diabetes is not known. Neutrophils contain large quantities of proteases, which they use to kill foreign invaders in the body. Neutrophils are known to release neutrophil elastase (NE) as a part of their response to injury, but failure to regulate their levels can result in tissue injury. We present evidence that neutrophil elastase plays and important role in the endothelial damage and cytotoxicity in diabetes, and postulate that the diabetes-induced induction of retinal inflammation and the vascular damage that is characteristic of early diabetic retinopathy are secondary to neutrophils via transport of NE in extracellular vesicles to endothelial cells, where the NE cleaves Gasdermin D (GSDMD) to cause cytotoxic pores in the endothelial cell membranes. We propose 3 specific aims: Aim 1. To investigate the effect of pharmacologic inhibition of NE on early stages of diabetic retinopathy. We will use structurally NE inhibitors, and will administer the therapies systemically as well as via eyedrops. Aim 2. To investigate mechanism(s) by which NE increases death of retinal endothelial cells in diabetes. Aim 3. Investigate the role of GSDMD in the pathogenesis of diabetic retinopathy. These studies will be conducted initially using GSDMD-/- mice. These studies will be conducted in vivo using pharmacological means to inhibit NE in diabetes and using mice genetically deficient in GSDMD. This proposal is novel because it focuses on (i) the role of a neutrophil protease in the pathogenesis of the retinopathy, and (ii) toxicity to retinal endothelial cells as a result of transfer of the protease from neutrophils to the endothelial cells via extracellular vesicles. This area is new and has not been previously been studied with respect to diabetic retinopathy. The insights learned from these studies can lead to development of novel and effective therapies that inhibit the development of diabetic retinopathy by targeting a protease secreted by neutrophils or the subsequent cleavage of GSDMD.

项目摘要现在大量数据表明，炎症过程在糖尿病发病机理中起关键作用视网膜病。尤其是白细胞在糖尿病引起的视网膜变性中起主要作用毛细血管（为最终发展视网膜缺血的情况设定了条件，释放了血管诸如VEGF和视网膜新血管形成之类的增殖因素）。白细胞如何介导糖尿病中的毛细血管细胞损伤尚不清楚。中性粒细胞含有大量蛋白酶，它们用来杀死体内外国入侵者。已知嗜中性粒细胞释放中性粒细胞弹性蛋白酶（NE）是其对损伤的反应的一部分，但未能调节其水平会导致组织损伤。我们提供了中性粒细胞弹性酶发挥作用的证据，并且在糖尿病的内皮损伤和细胞毒性中的重要作用，并假设糖尿病诱导诱导视网膜炎症和早期糖尿病性视网膜病特征的血管损伤是继发于中性粒细胞，通过在细胞外囊泡中转运到内皮细胞，NE裂解 Gasdermin D（GSDMD）在内皮细胞膜中引起细胞毒性孔。我们提出了3个具体目标：目的1。研究NE对NE对糖尿病性视网膜病早期阶段的药理抑制作用。我们将使用结构上的NE抑制剂，并将通过眼睛以及通过眼齿施用疗法。目的2。研究机制，通过该机制增加了糖尿病中视网膜内皮细胞死亡的死亡。目标3。研究GSDMD在糖尿病性视网膜病的发病机理中的作用。这些研究将是最初使用GSDMD - / - 小鼠进行。这些研究将使用药理手段在体内进行，以抑制糖尿病中的NE并使用小鼠 GSDMD遗传缺陷。该提议是新颖的，因为它重点放在（i）中性粒细胞的作用上视网膜病的发病机理中的蛋白酶，以及（ii）由于转移而对视网膜内皮细胞的毒性通过细胞外囊泡从中性粒细胞到内皮细胞的蛋白酶。这个区域是新的，没有先前已经研究过有关糖尿病性视网膜病。从这些研究中学到的见解可以导致开发新的有效疗法，从而抑制糖尿病性视网膜病的发展靶向由中性粒细胞分泌的蛋白酶或随后的GSDMD裂解。