Glucocorticoids, ocular hypertension and glaucoma

糖皮质激素、高眼压症和青光眼

基本信息

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
项目类别：
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

项目摘要

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治疗药物过多的疾病和状况。每年有超过 1% 的人口接受 GC 处方。尽管非常广泛和有效的抗炎作用，长期GC治疗会导致严重的副作用，包括对眼睛的伤害。接受长期 GC 治疗的个体中有 30-75% 出现 GC- 诱发性高眼压症 (OHT)，如果不加以识别，可能会导致医源性开角型青光眼永久性视力丧失。尽管认识到这种显着的 GC 副作用已超过 60 年，但我们仍然不了解 GC 诱导的 OHT 易感性差异的原因或机制负责 GC-OHT 的操作。我们之前已经证明，选择性剪接显性负性糖皮质激素受体 (GRb) 的亚型可抑制培养的人 TM 细胞中的 GC 活性。分离的TM细胞青光眼供体眼 (GTM) 的 GRb 水平较低，因此对 GC 更敏感。虽然一个许多研究已经检查了 TM 细胞和组织中 DEX 诱导的转录组，但没有发现表明哪些差异表达基因或分子途径参与 GC-OHT。一些研究表明，GC-OHT 的易感性是遗传性的，但目前还没有基因被改变。与 GC-OHT 明确相关。我们的总体假设是 GC-OHT 是： (a) 由以下比率决定： TM 中内源性 GRa 至 GRb 表达； (b) 由特定的分子途径介导区分 GC 响应者和非响应者眼睛； (c) 基因决定，以便 GC-OHT 基因可以被绘制和识别。这一总体假设将在 3 个具体目标中得到检验。具体目标#1：确定内源性 GRb 在人眼前节离体灌注中调节 GC-OHT 的作用培养和小鼠体内。具体目标 #2：确定 GC-OHT 耐药和 GC-OHT 中的 TM 转录组敏感品系小鼠和眼前段灌注培养人眼，以鉴定 GC-OHT 的分子途径。具体目标#3：绘制并识别基因负责使用 BXD 重组近交小鼠系的 QTL 进行 GC-OHT。这项研究具有创新性我们将评估内源性 GRb 在小鼠品系中的作用（使用我们的新小鼠 GC-OHT 模型）并在体外灌注培养对 GC-OHT 敏感性不同的人眼前节，使用小鼠菌株和人类灌注培养的前段对 GC-OHT 的反应不同从分子角度剖析负责 GC-OHT 的途径，并使用 BXD 重组体绘制 GC-OHT 基因图谱近交系小鼠。这项工作至关重要且意义重大，因为我们的实验结果将有助于确定内源性GRb在调节GC-OHT反应中的作用，其分子机制 GC-OHT，以及预测类固醇反应者的最佳和最有效的方法，这仍然是一个重要的未满足的问题随着 GC-OHT 的日益普遍，临床需求也随之增加。