The role of distal aqueous humor outflow tissue in glucocorticoid-induced glaucoma

远端房水流出组织在糖皮质激素诱发青光眼中的作用

• 批准号: 10667863
• 负责人: Weiming Mao
• 金额: $ 23.78万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667863
• 关键词: 3D Print; Anterior; Aqueous Humor; Area; Autoimmune Diseases; Bar Codes; Binding; Blindness; Cattle; Cell Nucleus; Cells; Clinical; Closure by clamp; Collagen; Complex; Cornea; Data; Deposition; Dexamethasone; Disease; Distal; Ethanol; Excision; Extracellular Matrix; Extravasation; Eye; Eye diseases; Fibronectins; Gene Expression; Glaucoma; Glucocorticoid Receptor; Glucocorticoids; Glues; Goals; Hematoxylin and Eosin Staining Method; Human; In Situ; In Situ Hybridization; Inflammatory; Keratoplasty; Knowledge; Left; Medicine; Methods; Mission; Modeling; Molecular; Mus; Ocular Hypertension; Oligonucleotides; Ophthalmologic Surgical Procedures; Pathway interactions; Patients; Perfusion; Phagocytes; Procedures; Protocols documentation; Public Health; Publishing; RNA; Reporting; Research; Resistance; Resolution; Retinal Ganglion Cells; Role; Sclera; Site; Slide; Spottings; Stains; Structure; Structure of sinus venosus of sclera; Technology; Thinness; Tissues; Trabecular meshwork structure; Veins; aqueous; cost; design; glaucoma surgery; interest; minimally invasive; novel; novel strategies; ocular hypotensive; responders and non-responders; response; tissue preparation; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Glucocorticoids are frequently used to treat inflammatory and autoimmune diseases. Although very effective, glucocorticoids may induce ocular hypertension (OHT)/glaucoma in some patients (they are called responders; the others are called non-responders) after prolonged topical or systemic use. The trabecular meshwork (TM) outflow pathway contains several tissues. From upstream to downstream of aqueous humor outflow, the tissues can be divided into proximal outflow tissues including the TM and Schlemm’s canal, as well as distal outflow tissues including the collector channel and aqueous vein. The importance of the distal flow tissue is not clear in glucocorticoid-induced OHT/glaucoma. Therefore, we will study our hypothesis that the distal outflow tissue, like the TM, also responds differentially to glucocorticoids and this response contributes to OHT. Our goal is to elucidate the molecular mechanism of glucocorticoid-induced OHT. We propose two specific aims. SA1. Determine if distal outflow tissues differentially regulate outflow resistance in GIG. Using our new perfusion culture model, we will perfuse paired human donor corneal rims in two study groups. Group a: Both rims will have the TM removed using the Kahook dual blade. One rim will be perfused with 0.1% ethanol (EtOH) as a vehicle control and the fellow rim with 100nM dexamethasone (DEX) for 7-10 days. IOP will be recorded and compared between eyes. At least 50 pairs of corneal rims will be studied. This group is used to determine if the distal outflow tissue is able to cause glucocorticoid-induced OHT. Group b: One rim will be perfused directly, while the fellow rim will have the TM tissue removed and then perfused. Both rims will be treated with DEX for 7-10 days and IOP will be recorded. At least 50 pairs of corneal rims will be studied. This group is used to determine if the distal outflow tissue is as important as the proximal tissue in glucocorticoid- induced OHT. SA2. Compare spatial gene expression between proximal and distal outflow tissues in GIG. Paired corneal rims with or without TM will be perfused with EtOH in one rim and DEX in the fellow rim. Both rims will be used for spatial gene expression studies. Tissues will be fixed and stained with hematoxylin and eosin to identify the outflow structure/region. Identified regions will be mounted on a chip on a slide. The chip contains 5000 bar-coded spots containing oligonucleotides for the capture, amplification, and identification of tissue RNA for RNA sequencing. The resolution of the chip is sufficient to study both proximal and distal outflow tissues. Both responder and non-responder tissues will be studied and compared. The results will be validated using in situ hybridization and immunostaining. Our project is novel because we will use the new corneal rim perfusion culture model and spatial gene expression technology. This study is significant because it will provide a novel treatment target, the distal outflow tissue, for treating glucocorticoid induced glaucoma.

项目摘要/摘要 糖皮质激素经常用于治疗炎症和自身免疫性疾病。尽管非常有效，但是 糖皮质激素可能会在某些患者中诱导眼高血压（OHT）/青光眼（称为反应者； 延长局部或全身使用后，其他人称为无反应者）。小梁网（TM） 出口途径包含多个组织。从上游到下游水性幽默出口， 组织可以分为包括TM和Schlemm的运河在内的近端出口组织，以及不同的 出口时机，包括收集器通道和水静脉。盘式流组织的重要性不是 在糖皮质激素诱导的OHT/青光眼中清除。因此，我们将研究远端出口的假设 像TM一样，组织对糖皮质激素的反应也有所不同，这种反应有助于OHT。我们的 目标是阐明糖皮质激素诱导的OHT的分子机制。我们提出了两个具体目标。 SA1。确定不同的出口组织是否对GIG中的出口阻力有所不同。使用我们的新 灌注培养模型，我们将在两个研究组中灌注配对的人角膜边缘。 A组：两者 轮辋将使用Kahook双刀片卸下TM。一个边缘将用0.1％乙醇灌注 （ETOH）作为媒介物控制，并用100nm地塞米松（DEX）进行7-10天的边缘。 IOP会 记录并比较眼睛之间。至少将研究50对角膜边缘。这个小组习惯了 确定远端出口组织是否能够引起糖皮质激素诱导的OHT。 B组：一个边缘将是 直接完善，同时轮辋将取出TM组织，然后灌注。两个轮辋将是 用DEX处理7-10天，将记录IOP。至少将研究50对角膜边缘。这 组用于确定远端出口组织是否与糖皮质激素中的代理组织一样重要 诱导的OHT。 SA2。比较近端和盘式出口正时之间的空间基因表达 演出。带有或不带有TM的配对角膜边缘将在一个边缘的一个边缘中灌注ETOH，并在同伴边缘中灌注。 两个轮辋将用于空间基因表达研究。组织将被固定并用苏木精染色 和曙红以识别出口结构/区域。确定的区域将安装在滑梯上的芯片上。这 芯片包含5000个含有寡核苷酸的条形码斑点，用于捕获，放大和识别 用于RNA测序的组织RNA。芯片的分辨率足以研究近端和不同 出口组织。响应者和非反应组织都将进行研究并进行比较。结果将是 使用原位杂交和免疫染色进行了验证。我们的项目是新颖的，因为我们将使用新的 角膜边缘灌注培养模型和空间基因表达技术。这项研究很重要，因为 它将提供一个新型的治疗靶标，即远端出口组织，用于治疗糖皮质激素诱导的青光眼。