Glucocorticoids, ocular hypertension and glaucoma

糖皮质激素、高眼压症和青光眼

• 批准号: 10468972
• 负责人: Abbot Frederick Clark
• 金额: $ 54.04万
• 依托单位: UNIVERSITY OF NORTH TEXAS HLTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10468972
• 关键词: Alternative Splicing; Anterior; Anti-Inflammatory Agents; Antiinflammatory Effect; Aqueous Humor; Biochemical; Bioinformatics; Biological; Biological Markers; Blindness; Cells; Chronic; Clinical; Cornea; Data; Development; Disease; Disease Resistance; Dominant-Negative Mutation; Eye; Generations; Genes; Glaucoma; Glucocorticoid Receptor; Glucocorticoids; Human; Iatrogenesis; Immunosuppressive Agents; Inbred Mouse; Individual; Inherited; Lead; Link; Maps; Mediating; Mediator of activation protein; Molecular; Mouse Strains; Mus; Ocular Hypertension; Open-Angle Glaucoma; Pathway interactions; Patients; Perfusion; Pharmacology; Phenotype; Physiologic Intraocular Pressure; Play; Population; Predisposition; Primary Open Angle Glaucoma; Protein Isoforms; Quantitative Trait Loci; RNA Interference; Recombinants; Reproducibility; Research; Resistance; Resistant Hypertension; Retinal Diseases; Risk Factors; Role; Spliceosomes; Steroids; Testing; Therapeutic Agents; Tissues; Trabecular meshwork structure; Transactivation; Work; aqueous; clinically relevant; clinically significant; differential expression; ex vivo perfusion; gene therapy; in vivo; innovation; knock-down; man; molecular phenotype; mouse model; pressure; receptor expression; responders and non-responders; response; side effect; therapy development; transcriptome; transcriptome sequencing

Glucocorticoids (GCs) are commonly used anti-inflammatory and immunosuppressive therapeutic agents for a plethora of diseases and conditions. Over 1% of our population receives GC prescriptions annually. Despite the very broad and potent anti-inflammatory effects, prolonged GC therapy can cause serious side effects, including damage to the eye. Between 30-75% of individuals receiving prolonged GC therapy develop GC- induced ocular hypertension (OHT), which if unrecognized can lead to iatrogenic open-angle glaucoma and permanent vision loss. Despite recognition of this significant GC side effect for more than six decades, we still do not understand the reason for differences in susceptibility to GC-induced OHT or the mechanism(s) of action responsible for GC-OHT. We have previously shown that the alternative spliced dominant negative isoform of the glucocorticoid receptor (GRb) inhibits GC activity in cultured human TM cells. TM cells isolated from glaucoma donor eyes (GTM) have low GRb levels and are therefore more sensitive to GCs. Although a number of studies have examined the DEX-induced transcriptome in TM cells and tissues, there is no indication which of the differentially expressed genes or molecular pathways are involved in GC-OHT. Several studies have shown that susceptibility to develop GC-OHT is genetically inherited, but no genes have been definitively linked to GC-OHT. Our overall hypothesis is that GC-OHT is: (a) determined by the ratio of endogenous GRa to GRb expression in the TM; (b) mediated by specific molecular pathways that can be differentiated from GC-responder and non-responder eyes; and (c) genetically determined so that GC-OHT genes can be mapped and identified. This overall hypothesis will be tested in 3 specific aims. Specific Aim #1: Determine the role of endogenous GRb in regulating GC-OHT in human anterior segment ex vivo perfusion culture and in vivo in mice. Specific Aim #2: Determine the TM transcriptome in GC-OHT resistant and sensitive strains of mice and in anterior segment perfusion cultured human eyes in order to identify the molecular pathways that are responsible for GC-OHT. Specific Aim #3: Map and identify the genes responsible for GC-OHT using QTL of the BXD recombinant inbred mouse lines. This research is innovative in that we will evaluate the role of endogenous GRb in mouse strains (with our new mouse model of GC-OHT) and in ex vivo perfusion cultured human anterior segments that differ in sensitivity to GC-OHT, use mouse strains and human perfusion cultured anterior segments that are differentially responsive to GC-OHT to molecularly dissect the pathway responsible for GC-OHT, and map GC-OHT genes using BXD recombinant inbred mice. This work is essential and significant because our experimental results will help determine the role of endogenous GRb in regulating responsiveness to GC-OHT, the molecular mechanisms responsible for GC-OHT, and the best and most effective way to predict steroid responders, which still is an important unmet clinical need as GC-OHT is becoming increasingly prevalent.

糖皮质激素（GC）是常用的抗炎和免疫抑制剂治疗剂 大量疾病和条件。我们超过1％的人每年收到GC处方。尽管 长时间的GC治疗非常广泛，有效的抗炎作用，可能会引起严重的副作用， 包括对眼睛的损害。在接受长期GC治疗的人中，有30-75％的人发展了GC- 诱导眼高血压（OHT），如果未被认可，可能会导致医源性开式角膜素质青光眼和 永久视力丧失。尽管认识到六十多年来这种重要的GC副作用，但我们仍然 不了解GC引起的OHT敏感性差异或机制的敏感性差异的原因 负责GC-OHT的行动。我们先前已经表明，替代剪接的主要负面 糖皮质激素受体（GRB）的同工型抑制培养的人TM细胞中的GC活性。 TM细胞分离 来自青光眼供体眼（GTM）的GRB水平较低，因此对GC更敏感。虽然 研究数量已经检查了DEX诱导的TM细胞和组织中的转录组，没有 指示差异表达的基因或分子途径参与GC-OHT。一些 研究表明，开发GC-OHT的易感性是遗传遗传的，但没有基因是 最终与GC-OHT链接。我们的总体假设是GC-OHT是：（a）由 内源性GRA到TM中的GRB表达； （b）通过特定的分子途径介导的 与GC响应者和无反应的眼睛区分开； （c）遗传确定的GC-OHT 基因可以映射和鉴定。该总体假设将以3个特定目的进行检验。特定目标＃1： 确定内源性GRB在调节GC-OHT人体前部灌注中的作用 培养和体内小鼠。特定目的＃2：确定GC-OHT抗性和 小鼠敏感菌株和前部灌注培养的人眼，以识别 负责GC-OHT的分子途径。特定目的＃3：地图并识别基因 使用BXD重组近交小鼠系的QTL负责GC-OHT。这项研究是创新的 因为我们将评估内源性GRB在小鼠菌株中的作用（使用我们的新小鼠GC-OHT模型） 在体内灌注中培养的人体前部，对GC-OHT的敏感性有所不同，请使用小鼠 菌株和人类灌注对GC-OHT有差异响应的前部培养的前部培养 分子剖析负责GC-OHT的途径，并使用BXD重组绘制GC-OHT基因 近交小鼠。这项工作至关重要，因为我们的实验结果将有助于确定 内源性GRB在调节对GC-OHT的反应性中的作用，该分子机制负责 GC-OHT，也是预测类固醇响应者的最佳，最有效的方法，这仍然是重要的未满足 随着GC-OHT的临床需求越来越普遍。