Transfer: Molecular Mechanisms of Glaucoma

转：青光眼的分子机制

• 批准号: 10246532
• 负责人: Dorota Skowronska-Krawczyk
• 金额: $ 38.07万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10246532
• 关键词: 9p21; ATAC-seq; Adult; Affect; African American; Age; Animal Model; Bilateral; Biological Models; Blindness; CDKN2A gene; CRISPR/Cas technology; Categories; Cell Aging; Cell Death; ChIP-seq; Code; Complex; Development; Disease; Disease Progression; Down-Regulation; Engineering; Environmental Risk Factor; Epigenetic Process; Etiology; Eye; Eye Development; Future; Gene Expression Regulation; Genes; Genetic; Genetic Risk; Genomics; Glaucoma; Human; Human Chromosomes; Inherited; Laboratories; Laboratory mice; Linkage Disequilibrium; Mediating; Methodology; Modeling; Molecular; Molecular Analysis; Mouse Strains; Mus; Nerve Degeneration; Nucleic Acid Regulatory Sequences; Optic Nerve; Pathogenesis; Pathology; Patients; Physiologic Intraocular Pressure; Population; Positioning Attribute; Primary Open Angle Glaucoma; Quality of life; Research; Research Personnel; Retina; Retinal Ganglion Cells; Risk; Risk Factors; Role; Sampling; Signal Transduction; Technology; Testing; Tumor Suppressor Proteins; United States; Up-Regulation; Variant; Visual impairment; Work; age related; blind; design; epigenomics; genetic association; genomic locus; human tissue; humanized mouse; improved; mouse model; nerve damage; novel; optic nerve disorder; personalized therapeutic; protective effect; retinal ganglion cell degeneration; risk variant; senescence; therapeutic development; transcriptomics

SUMMARY Glaucoma is a group of optic neuropathies characterized by slow, progressive loss of retinal ganglion cells (RGCs), optic nerve degeneration and as a consequence, vision loss. It has been estimated that more than 70 million people are currently affected by glaucoma with approximately 10% being bilaterally blind, making it the leading cause of irreversible blindness in the world. Several glaucoma categories exist, but in United States, most of the cases are primary open-angle glaucoma (POAG), a variant particularly prevalent amongst African Americans. POAG is recognized as a complex disease in which multiple genetic and environmental factors interact. The two leading risk factors, increase intraocular pressure (IOP) and age are related to the extent and rate of RGC loss. Recent advances in genomics have allowed researchers to describe genetic association between the risk of glaucoma and specific genomic loci. Nevertheless, despite years of research, the molecular basis of glaucoma is poorly understood and the factors contributing to its progression have not been fully characterized. In our recent work, we have used a mouse model to study the molecular impact of Six6 risk variant in development of glaucoma and in RGC death. We observed that upon increased IOP, expression of Six6 increases and directly regulates the expression of p16Ink4a, leading to enhanced senescence in RGCs and most likely directly causing RGC death. The gene encoding p16INK4a, CDKN2A, lies within the tumor suppressor locus on human chromosome 9p21. This locus has been independently identified by several groups to have the highest association with POAG in different population samples. Gene regulation within the 9p21 locus has been extensively studied in many laboratories; however, a molecular analysis has never been performed specifically in relation to glaucoma. Mouse Six6 harbors His at position 141 and therefore is ideal to investigate the molecular role of this variant in glaucoma. However, due to the lack of non-risk variant in mouse strains, it is not possible to study the contribution of both variants on RGC development and degeneration in a mouse model. Here, we propose to use CRISPR/Cas9 technology to engineer mice harboring human non-risk variant of SIX6 to study the impact of each variant in pathogenesis of glaucoma. We will use state-of-the art molecular and cellular technologies to study retinal development and RGC degeneration upon elevated intraocular pressure as a function of the particular variant of SIX6. In addition, we will investigate the molecular mechanisms of p16Ink4a upregulation in the etiology of the disease using transcriptomic and epigenomic approaches, and we will propose the methodology to downregulate its expression in the eye. The proposed combination of approaches will move forward the general understanding of the etiology of glaucoma and provide the molecular basis for development of novel, personalized, therapeutic strategies to improve the quality of life for glaucoma patients.

总结 青光眼是一组以视网膜神经节细胞缓慢、进行性丧失为特征的视神经病变 视网膜神经节细胞（RGC），视神经变性，并因此导致视力丧失。据估计，超过70 目前有100万人患有青光眼，其中约10%为双眼失明， 导致世界上不可逆失明的主要原因。存在几种青光眼类别，但在美国， 大多数病例是原发性开角型青光眼（POAG）， 美国人原发性开角型青光眼是一种复杂的疾病， 互动.两个主要的危险因素，眼内压（IOP）升高和年龄与程度和 研资局的损失率。基因组学的最新进展使研究人员能够描述遗传关联 青光眼风险和特定基因位点之间的联系。然而，尽管经过多年的研究， 青光眼的基础知之甚少，导致其进展的因素尚未完全了解。 表征了在我们最近的工作中，我们使用小鼠模型来研究Six 6风险的分子影响。 在青光眼的发展和RGC死亡中的变异。我们观察到，在IOP升高时， Six 6增加并直接调节p16 Ink 4a的表达，导致RGC衰老增强 很可能直接导致皇家海军陆战队的死亡编码p16 INK 4a的基因CDKN 2A位于肿瘤内。 人类染色体9 p21上的抑制基因座。这个基因座已经被几个研究小组独立鉴定 在不同人群样本中与POAG的相关性最高。9 p21内的基因调控 基因座已经在许多实验室中被广泛研究;然而，分子分析从未被 专门用于治疗青光眼。小鼠Six 6在位置141处具有His，因此是理想的 研究这种变异在青光眼中的分子作用。然而，由于小鼠中缺乏非风险变体， 菌株，这是不可能的，研究的贡献，这两个变种对RGC的发展和退化， 小鼠模型在这里，我们建议使用CRISPR/Cas9技术来工程小鼠， SIX 6的变体，以研究每个变体在青光眼发病机制中的影响。我们将使用最先进的 分子和细胞技术，研究视网膜发育和RGC变性后， 眼内压作为SIX 6的特定变体的函数。此外，我们将研究分子 使用转录组学和表观基因组学研究p16 Ink 4a上调在疾病病因学中的机制 方法，我们将提出下调其在眼睛中表达的方法。拟议 这些方法的结合将推进对青光眼病因学的一般理解，并提供 开发新的、个性化的治疗策略以改善生活质量的分子基础 对于青光眼患者。