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

• 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
• 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

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.

项目概要 现在大量数据表明炎症过程在糖尿病的发病机制中发挥着关键作用 视网膜病变。尤其是白细胞似乎在糖尿病引起的视网膜变性中发挥着重要作用 毛细血管（为视网膜缺血的最终发展、释放血管- VEGF 等增殖因子，最终导致视网膜新生血管形成）。白细胞如何介导这一过程 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的蛋白酶，它们用来杀死体内的外来入侵者。 众所周知，中性粒细胞会释放中性粒细胞弹性蛋白酶（NE），作为其对损伤反应的一部分，但未能 调节它们的水平会导致组织损伤。我们提供的证据表明中性粒细胞弹性蛋白酶发挥作用 在糖尿病的内皮损伤和细胞毒性中发挥重要作用，并假设糖尿病引起的 诱发视网膜炎症和血管损伤是早期糖尿病视网膜病变的特征 通过将细胞外囊泡中的 NE 转运至内皮细胞，继发于中性粒细胞，并在此处裂解 Gasdermin D (GSDMD) 在内皮细胞膜上产生细胞毒性孔。 我们提出3个具体目标： 目的 1. 探讨药物抑制 NE 对早期糖尿病视网膜病变的影响。我们 将使用结构性 NE 抑制剂，并通过滴眼剂和全身给药进行治疗。 目标 2. 研究 NE 增加糖尿病视网膜内皮细胞死亡的机制。 目标 3. 研究 GSDMD 在糖尿病视网膜病变发病机制中的作用。这些研究将 最初使用 GSDMD-/- 小鼠进行。 这些研究将在体内使用药理学手段抑制糖尿病中的 NE 并使用小鼠进行 GSDMD 遗传缺陷。该提案很新颖，因为它重点关注（i）中性粒细胞的作用 蛋白酶在视网膜病发病机制中的作用，以及 (ii) 转移对视网膜内皮细胞的毒性 蛋白酶从中性粒细胞通过细胞外囊泡到达内皮细胞。这个区域是新的，没有 以前曾对糖尿病视网膜病变进行过研究。从这些研究中获得的见解可以 导致开发出新颖有效的疗法，通过以下方式抑制糖尿病视网膜病变的发展 靶向中性粒细胞分泌的蛋白酶或随后的 GSDMD 裂解。