Elucidating novel mechanisms for glucocorticoid-induced ocular hypertension

项目摘要/摘要 背景信息和相关性：糖皮质激素(GC)广泛应用于眼科和 一般的医学。GC诱导的高眼压(OHTN)是一种常见的并发症，可 可能会让人失明。玻璃体内注射GC的这种并发症的发生率在28-34%之间。 取决于注射的配方。这种并发症的风险因素尚不清楚，但有 长期存在的证据表明遗传因素起到了作用。几乎没有进行过基因研究来确定 被牵连的变种和没有一致的发现还没有确定。假设：遗传易感性 GC诱导的OHTN既有常见的变异，也有罕见的变异，相关的变异更有可能是 在转录本因接触GC而改变的基因以及具有糖皮质激素反应的基因中发现 元素(GRE)或与原发性开角型青光眼(POAG)或眼压有关 (眼压)。具体目标：1.人眼小梁网(TM)细胞的GC转录组 Schlemm氏管内皮细胞。2.进行全基因组基因分型和整个外显子组 1086例玻璃体内注射糖皮质激素暴露患者的序列分析3.辨别常见和罕见 编码区和非编码区的遗传变异与GC诱导的眼压升高有关。 方法：我们将人TM和SCE细胞暴露于地塞米松和培养液对照中，并进行 RNA测序以确定受GC转录调控的基因。我们将在全基因组范围内进行 1086例连续眼压患者玻璃体内注射GC后的基因分型和WES 测量。基因分型和WES将作为一种具有成本效益的分析方法 建议。我们将研究GC诱导的OHTN作为调整后的眼压变化的数量性状 基线眼压和一个二分特征(GC响应者与非响应者)。我们将在全基因组范围内执行 关联分析和WES分析以及目标分析。我们将以基因的变异为目标 在带有GREs的基因座和先前相关的基因中，受目标1中确定的GCs转录调控 患有眼压或开角型青光眼。我们将检查同一基因座上的罕见变异与疾病结局的关系 集料和负担测试。我们将招募530名参与者的复制队列，并有针对性地执行 对发现分析中的提示性或有意义的发现进行后续排序。我们还将在 用现有的基因组数据在三个外部队列中重复这些发现。暗示：如果基因 与GC诱导的眼压升高相关，他们不仅将提供对这种并发症的洞察 病理生理学，也更广泛地转化为POAG的机制。我们的发现将对临床产生影响。 GC患者的管理。如果我们能够识别出有GC诱导性OHTN风险的患者，我们可能会避免 在患者接受GC治疗期间，通过使用替代疗法或增加监测来增加发病率。

Project Summary/Abstract Background Information and Relevance: Glucocorticoids (GC) are widely used in ophthalmology and in medicine in general. GC-induced ocular hypertension (OHTN) is a common complication that can be potentially blinding. The rate of this complication for GC injected intravitreally varies between 28-34% depending on the formulation injected. This complication's risk factors are not well understood but there is long-standing evidence that genetic factors play a role. Few genetic studies have been undertaken to identify the implicated variants and no consistent findings have been identified. Hypotheses: Genetic susceptibility to GC-induced OHTN is due to both common and rare variation, and associated variants are more likely to be found in genes with transcripts altered by exposure to GC as well as genes that have glucocorticoid response elements (GREs) or have been associated with primary open angle glaucoma (POAG) or intraocular pressure (IOP). Specific Objectives: 1.To characterize the GC transcriptome in human trabecular meshwork (TM) cells and Schlemm's canal endothelial (SCE) cells. 2. To perform genome-wide genotyping and whole exome sequencing (WES) on 1086 patients with intravitreal GC injection exposure. 3. To identify common and rare genetic variants in both coding and non-coding regions that are associated with GC-induced IOP elevation. Methods: We will expose human TM and SCE cells to dexamethasone as well as media controls and perform RNA sequencing to identify genes that are transcriptionally regulated by GCs. We will perform genome-wide genotyping and WES on 1086 patients who have received an intravitreal GC injection and have serial IOP measurements. Genotyping and WES will be performed as a cost efficient approach for the analyses proposed. We will examine the GC-induced OHTN both as a quantitative trait of change in IOP adjusted for baseline IOP and a dichotomous trait (GC responders vs. non-responders). We will execute genome-wide association and WES analyses as well as targeted analyses. We will target variants in genes that are transcriptionally regulated by GCs as identified in Aim 1, in loci with GREs, and in genes previously associated with IOP or POAG. We will examine rare variants in the same loci for association with disease outcome using aggregate and burden tests. We will enroll a replication cohort of 530 participants and perform targeted sequencing to follow up suggestive or significant findings from the discovery analyses. We will also perform in silico replication of these findings in three external cohorts with existing genomic data. Implications: If genes associated with GC-induced IOP rise are identified, they will not only provide insights into this complication's pathophysiology, but also into POAG mechanisms more broadly. Our findings will have implications for clinical management of patients on GC. If we are able to identify patients at risk of GC-induced OHTN, we may avoid morbidity by using alternative therapies or increasing surveillance while patients receive GC.