Retina-derived extracellular vesicles in diabetic retinopathy: their potential role in pathogenesis and therapy

糖尿病视网膜病变中视网膜来源的细胞外囊泡：它们在发病机制和治疗中的潜在作用

• 批准号: 10644819
• 负责人: Emma M Lessieur
• 金额: $ 10.76万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644819
• 关键词: Adherence; Adhesions; Affect; Anatomy; Animals; Area; Binding; Blindness; Blood Vessels; Blood capillaries; Cell Adhesion; Cell Adhesion Molecules; Cell Communication; Cell Line; Cell Surface Receptors; Cells; Characteristics; Chronic; Clinical; Complication; Complications of Diabetes Mellitus; Data; Development; Diabetes Mellitus; Diabetic Retinopathy; Diabetic mouse; Disease; Disease Progression; Endothelial Cells; Etiology; Eukaryotic Cell; Exposure to; Eyedrops; Foundations; In Vitro; Inflammation; Inflammatory; Injections; Integral Membrane Protein; Intercellular adhesion molecule 1; Investigation; Learning; Leukocytes; Light; Mediating; Methodology; Methods; Modeling; Molecular; Mus; Neural Retina; Parents; Pathogenesis; Pathologic; Pathology; Permeability; Phenotype; Photoreceptors; Physiological; Plasma; Preparation; Process; Proteins; Protocols documentation; Publishing; Receptor Cell; Reporter; Reporting; Research; Retina; Retinal Diseases; Rhodopsin; Rod; Role; Route; Secondary to; Sorting; Strategic Planning; Structure; Surface; Systemic Therapy; TLR4 gene; Tail; Testing; Tissues; Transgenic Mice; Transgenic Organisms; Up-Regulation; Vascular Diseases; Vascular Endothelium; Veins; Visual impairment; Work; aging population; antagonist; cytotoxicity; diabetic; experimental study; extracellular vesicles; in vivo; inhibitor; insight; intravitreal injection; leukocyte activation; monolayer; mouse model; multiple omics; mutant mouse model; nanoscale; non-diabetic; novel; novel therapeutic intervention; pharmacologic; post-doctoral training; receptor; targeted treatment; vesicular release; virtual; visual cycle

Project summary. Extracellular vesicles are cell-derived membranous structures harboring a variety of biomolecules. Their secretion and cargo are determined in part by the physiological or pathological conditions that the parent cell is exposed to. Recently, increased levels of extracellular vesicles in plasma have been associated with the pathogenesis of diabetic retinopathy, but from where those plasma extracellular vesicles originated from, and whether they affect the development of the disease is still under investigation. Early in my postdoctoral training, I established a modified protocol to isolate extracellular vesicles from small tissues, including retina. Using this protocol, I isolated extracellular vesicles of retina explants from nondiabetic and diabetic mice and found that diabetic retina-derived extracellular vesicles induce ICAM-1, an important adhesion molecule, in retinal endothelial cells in vitro. This induction is inhibited by pharmacological blockage of TLR4, a transmembrane protein that has been proposed as an extracellular vesicles cell receptor. I also found that diabetic retina-derived extracellular vesicles activate circulating leukocytes isolated from nondiabetic animals. This work laid the foundation of my ongoing research. I want to determine the mechanism(s) of how retina-derived extracellular vesicles induce ICAM-1 in retinal endothelial cells and activate circulating leukocytes using a mouse model of diabetes. I propose 3 specific aims: Aim 1, To determine if a diabetes-modified visual cycle activity in photoreceptor cells modulates the retina- derived extracellular vesicles phenotype, which lead to the induction of ICAM-1 in retinal endothelial cells and activation of circulating leukocytes. Aim 2, To determine if TLR4 is the common receptor present in retinal endothelial cells and leukocytes and is triggered upon binding of retina-derived extracellular vesicles, leading to the induction of ICAM-1 in retinal endothelial cells and activation of circulating leukocytes in diabetes. Aim 3, To determine if rod-derived extracellular vesicles are key contributors to the retinal vascular pathology characteristic of diabetic retinopathy. These studies will be conducted in vivo using mice genetically deficient in key proteins of the visual cycle and pharmacological means to inhibit extracellular vesicles secretion and TLR4 receptor in diabetes. This proposal is novel because it focuses on (i) the role of retina (and rod)-derived extracellular vesicles in the pathogenesis of the retinopathy, (ii) the role of the visual cycle in the cargo sorting of retina (and rod)-derived extracellular vesicles, and (iii) the role of TLR4 as a receptor of retina (and rod)-derived extracellular vesicles. The insights learned from these studies can lead to the development of novel and effective targeted therapies to inhibit the etiopathogenesis of diabetic retinopathy.

项目摘要。 细胞外囊泡是细胞衍生的膜结构，含有多种生物分子。他们的 分泌物和货物部分地由亲本细胞所处的生理或病理条件决定。 暴露于。最近，血浆中细胞外囊泡水平的增加与糖尿病的发生有关。 糖尿病视网膜病变的发病机制，但从哪里这些血浆细胞外囊泡起源， 是否影响病情发展，仍在调查中。 在我博士后培训的早期，我建立了一个改进的方案，从小细胞中分离细胞外囊泡， 组织，包括视网膜。使用这个协议，我分离的视网膜外植体细胞外囊泡从非糖尿病 和糖尿病小鼠，发现糖尿病视网膜衍生的细胞外囊泡诱导ICAM-1，一个重要的 粘附分子，在体外视网膜内皮细胞。这种诱导作用可通过药理学阻断 TLR 4是一种跨膜蛋白，被认为是细胞外囊泡细胞受体。我还发现 糖尿病视网膜衍生的细胞外囊泡激活从非糖尿病视网膜分离的循环白细胞， 动物这项工作奠定了我正在进行的研究的基础。我想确定的机制（S）如何 视网膜衍生的细胞外囊泡诱导视网膜内皮细胞中的ICAM-1并激活循环白细胞 使用的是糖尿病小鼠模型。 我提出三个具体目标： 目的1，确定糖尿病改变的感光细胞视觉周期活动是否调节视网膜- 衍生的细胞外囊泡表型，其导致视网膜内皮细胞中ICAM-1的诱导， 激活循环白细胞。 目的2，确定TLR 4是否是视网膜内皮细胞和白细胞的共同受体， 在视网膜衍生的细胞外囊泡结合后触发，导致视网膜中ICAM-1的诱导。 糖尿病中内皮细胞和循环白细胞的激活。 目的3，确定视杆细胞源性细胞外囊泡是否是视网膜血管病变的关键因素 糖尿病视网膜病变的特征。 这些研究将在体内使用视觉周期关键蛋白质遗传缺陷的小鼠进行， 药理学手段抑制糖尿病细胞外囊泡分泌和TLR 4受体。这项建议 是新颖的，因为它集中于（i）视网膜（和视杆）衍生的细胞外囊泡在发病机制中的作用 的视网膜病变，（ii）的作用，视觉周期中的货物分类视网膜（和杆）衍生的细胞外 囊泡，和（iii）TLR 4作为视网膜（和杆）衍生的细胞外囊泡的受体的作用。的见解 从这些研究中了解到的信息可以导致新的和有效的靶向治疗的发展，以抑制 糖尿病视网膜病变的发病机制。