Mechanisms of lens epithelium fibrosis and its relevance to posterior capsule opacification

晶状体上皮纤维化的机制及其与后囊膜混浊的相关性

• 批准号: 10685531
• 负责人: Xingjun Fan
• 金额: $ 38.5万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10685531
• 关键词: Address; Affect; Animal Model; Aqueous Humor; Autocrine Communication; Biogenesis; Biology; Cataract; Cataract Extraction; Cell Communication; Cell Culture Techniques; Chronic; Chronic Disease; Colorado; Communication; Complication; Data; Disease; Endocrine; Epithelial Cell Proliferation; Epithelial Cells; Epithelium; Fibrosis; Human; In Vitro; Inflammation; Inflammatory; Investigation; Kinetics; Light; Mediating; Mus; Operative Surgical Procedures; Oryctolagus cuniculus; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Physiological; Pilot Projects; Play; Primary Cell Cultures; Process; Proliferating; Recording of previous events; Regulatory Pathway; Research; Residual state; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Stimulus; TNF gene; Testing; Transforming Growth Factor beta; Universities; Visual Acuity; WNT Family Gene; WNT Signaling Pathway; beta catenin; capsule; cell growth; cohort; cytokine; epithelial to mesenchymal transition; exosome; extracellular vesicles; high risk; human tissue; in vivo; in vivo Model; lens; migration; mouse model; novel; oxidation; prevent; therapeutic target; trafficking; transcriptome sequencing; transdifferentiation

Abstract Cataract surgery is associated with a high risk of posterior capsule opacification (PCO), a form of secondary cataract, the formation of which is still poorly understood. However, PCO is generally considered a chronic disease and a consequence of the LEC fibrotic process, attributed in part to the epithelial-mesenchymal transition (EMT) in which TGFβ mediated signaling is one of the key players. Recently, we and others have suggested that canonical Wnt/β-catenin signaling is also a pivotal player in LEC fibrosis and the pathogenesis of PCO. To test the Wnt signaling in chronic conditions, we conducted a 6-month-kinetic rabbit cataract surgery study and found a profound activation of the canonical Wnt signaling during the course of PCO formation, associated with elevated oxidation and chronic inflammation. We postulate that lens epithelial cells (LECs) undergo self- regulatory signaling transduction to adapt to the new microenvironment after cataract surgery, and the canonical Wnt/β-catenin plays a pivotal role in this process. Our initial study identifies Wnt3 as a highly expressed and upregulated Wnt-family gene in human and mouse LECs after cataract surgery. However, the role of Wnt3 signaling mediated LEC fibrosis and PCO formation is still unclear; the underlying mechanisms of how lens epithelial cells regulate the Wnt signaling remain to be addressed. We plan to unravel these puzzles with a basis of support from several key findings of our pilot studies, e.g., our discovery that LECs package Wnt3 into exosomes, a type of extracellular vesicle (EVs), and conduct signaling transduction in an endocrine fashion. Importantly, inflammatory cytokines, such as TNFα, can drastically promote exosome biogenesis and Wnt3 secretion. Aim 1 will test how lens epithelial cells regulate Wnt3 packaging and exosome biogenesis. We need to understand how LECs manage to promote exosome biogenesis and Wnt3 secretion under the certain stimuli, e.g., inflammatory cytokines and oxidative stress. Aim 2 will delineate the role of Wnt3 in LEC proliferation, migration, and differentiation using in vitro cell culture, primary cell culture, ex vivo explant culture, and in vivo mouse models. Aim3 studies the LECs’ growth microenvironment and the Wnt3 mediated autocrine signaling by using human tissue with and without a history of cataract surgery. Human tissue studies will offer a final checkpoint of research after studies from cell culture and animal models. In summary, substantial pilot data form the basis of our proposal, one which will unravel a novel LECs self-regulatory pathway that is closely reminiscent of the pathological and physiological conditions of cataract surgery.

抽象的 白内障手术与后胶囊不透明（PCO）的高风险有关，这是一种次级形式 白内障，其形成仍然很少了解。但是，PCO通常被认为是慢性 疾病和LEC纤维化过程的结果，部分归因于上皮 - 间质转变 （EMT）TGFβ介导的信号传导是关键参与者之一。最近，我们和其他人建议 规范的Wnt/β-catenin信号传导也是LEC纤维化和PCO发病机理的关键参与者。到 在慢性条件下测试Wnt信号传导，我们进行了6个月的运动兔白内障手术研究和 在PCO形成过程中发现了典型的Wnt信号传导的深刻激活，与 氧化和慢性炎症升高。我们假设晶状体上皮细胞（LEC）发生自我 监管信号转换，以适应白内障手术后的新微环境和规范 Wnt/β-catenin在此过程中起关键作用。我们的初步研究将WNT3确定为高度表达的，并且 白内障手术后，在人和小鼠LEC中上调Wnt家庭基因。但是，WNT3的作用 信号传导介导的LEC纤维化和PCO形成尚不清楚。镜片的基本机制 调节Wnt信号的上皮细胞仍有待解决。我们计划以基础来解开这些难题 我们的飞行员研究的几个关键发现的支持，例如，我们发现LECS软件包Wnt3进入 外泌体，一种细胞外囊泡（EV），并以内分泌方式进行信号传导翻译。 重要的是，炎性细胞因子（例如TNFα）可以大大促进外泌体生物发生和Wnt3 分泌。 AIM 1将测试透镜上皮细胞如何调节Wnt3包装和外泌体生物发生。我们需要 要了解LEC如何在某些刺激下促进外泌体生物发生和Wnt3分泌， 例如，炎性细胞因子和氧化应激。 AIM 2将描述Wnt3在LEC增殖中的作用， 使用体外细胞培养，原代细胞培养，离体外植体和体内的迁移和分化 鼠标模型。 AIM3研究LEC的生长微环境和Wnt3介导的自分泌信号传导 使用有或没有白内障手术史的人体组织。人体组织研究将提供最终 从细胞培养和动物模型进行研究后的研究检查点。总之，大量的试点数据表 我们的提议的基础，该提案将揭示一个新颖的LEC自我调节途径，这是密切提醒的 白内障手术的病理和身体状况。