Validating novel candidate genes for normal-tension glaucoma

验证正常眼压性青光眼的新候选基因

• 批准号: 10749126
• 负责人: William Thomas Presley
• 金额: $ 4.08万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10749126
• 关键词: 12q13; ATP phosphohydrolase; Adult; Affect; Biological Assay; Blindness; Blood; Blood specimen; Candidate Disease Gene; Cells; Chromosomes; Classification; Co-Immunoprecipitations; Code; Coupling; Data; Databases; Defect; Diagnosis; Disease; Disease Progression; Early treatment; Electrostatics; Escherichia coli; Extracellular Matrix; Extracellular Matrix Proteins; Family; Family member; Functional disorder; Future; Gene Expression; Gene Expression Profiling; Gene Mutation; Genes; Genetic; Genetic Diseases; Genomics; Glaucoma; Goals; Heat-Shock Proteins 90; Heritability; Impairment; In Vitro; Individual; Link; Maintenance; Mendelian disorder; Methods; Missense Mutation; Modeling; Molecular; Molecular Chaperones; Mutation; N-terminal; Normal Range; Nucleotides; Optic Atrophy; Optic Nerve; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Physiologic Intraocular Pressure; Population; Positioning Attribute; Prevalence; Production; Proteins; RNA Polymerase II; Recording of previous events; Regulation; Risk; Risk Factors; Role; Running; Scaffolding Protein; Scientist; Structure; Tertiary Protein Structure; Testing; Therapeutic; Tissue-Specific Gene Expression; Variant; autosome; biobank; career; cohort; early screening; exome; experimental study; gene discovery; gene function; genetic pedigree; genetic variant; genome analysis; genome sequencing; genome wide association study; human disease; improved; in silico; member; modifiable risk; molecular diagnostics; mutant; novel; novel therapeutics; optic nerve disorder; protein protein interaction; rare variant; retinal ganglion cell degeneration; risk variant; scaffold; screening; segregation; skills; traditional therapy; transcriptome sequencing; whole genome

ABSTRACT My ultimate career goal is to become an independent scientist researching the genetic mechanisms of Mendelian disorders in hopes of improving patient therapeutics. Specifically, I am focused on disease gene discovery in familial cohorts, which remains one of the most effective means by which to study normal- tension glaucoma (NTG). Glaucoma describes a group of ocular conditions characterized by the progressive degeneration of the retinal ganglion cells (RGCs) that comprise the optic nerve and is present in 1-3% of the adult population. Traditional therapies have focused on lowering intraocular pressure (IOP), the largest modifiable risk factor. However, a subset of individuals with NTG – a diagnosis which accounts for nearly one-third of all glaucoma cases and denotes optic atrophy despite an IOP that is within normal range – continue to show disease progression after IOP-lowering treatment. A critical barrier to developing new treatments is understanding the molecular pathogenesis of NTG. To this end, our group identified a pedigree with autosomal dominant NTG that is poorly responsive to IOP-lowering therapy. Linkage, whole-exome, and whole-genome analyses revealed that there is likely more than one risk allele of high effect segregating in individual family members, though the vast majority of cases seem to be explained by a very rare missense substitution at a highly conserved residue in scaffolding/chaperone protein RPAP3. This variant, with high in silico pathogenicity scores, is predicted to disrupt the N-terminal domain of the protein. The N-terminal domain is critical for stimulating the ATPase activity of HSP90, another scaffolding/chaperone protein that modulates the production of extracellular matrix (ECM) components. Further, RNA-seq from familial blood samples revealed that ECM-associated genes were among the most dysregulated in a differential gene expression analysis. These observations support the hypothesis that disruptions in the N-terminus of RPAP3 impair HSP90 regulation, leading to defects in ECM production/maintenance. This could result in normal-tension glaucoma by compromising key optic nerve-supporting structures like the lamina cribrosa. My proposal seeks to: (i) determine the functional consequences of RPAP3 coding variants on HSP90 interaction/stimulation and ECM gene expression, (ii) determine the prevalence of deleterious RPAP3 variants in a cohort of patients diagnosed with glaucoma, and (iii) elucidate additional glaucoma risk variants segregating in this large family. The results may lead to improved screening and treatment options for NTG patients. By carrying out these aims, I will also broaden my skillset in disease gene discovery and functional characterization of genetic variants, poising me for a career focused on human disease genetics and molecular diagnostics.

抽象的 我的最终职业目标是成为一名研究遗传机制的独立科学家 具体来说，我专注于疾病的研究，希望能够改善患者的治疗。 家族队列中的基因发现，这仍然是研究正常人群的最有效手段之一 张力性青光眼 (NTG) 描述了一组以以下特征为特征的眼部疾病。 构成视神经的视网膜神经节细胞 (RGC) 进行性变性 1-3% 的成年人群中存在这种情况，传统疗法的重点是降低眼内压。 (IOP)，最大的可改变的危险因素，然而，NTG 患者的一个子集——这一诊断 占所有青光眼病例的近三分之一，并且表示视神经萎缩，尽管眼压在 正常范围——降低眼压治疗后疾病继续进展，这是治疗的一个关键障碍。 开发新的治疗方法正在了解 NTG 的分子发病机制。为此，我们的团队。 确定了一个患有常染色体显性 NTG 的家系，该家系对降眼压治疗反应不佳。 连锁、全外显子组和全基因组分析表明，可能存在不止一种风险等位基因 尽管绝大多数情况似乎是对个别家庭成员进行高效隔离 解释为支架/分子伴侣中高度保守的残基上非常罕见的错义取代 蛋白质 RPAP3 具有较高的计算机致病性评分，预计会破坏 N 末端。 蛋白质的 N 末端结构域对于刺激 HSP90 的 ATP 酶活性至关重要， 另一种调节细胞外基质 (ECM) 产生的支架/伴侣蛋白 此外，家族血液样本的 RNA-seq 揭示了 ECM 相关基因。 这些观察结果支持差异基因表达分析中最失调的。 假设 RPAP3 N 末端的破坏会损害 HSP90 的调节，从而导致缺陷 在 ECM 生产/维护中，这可能会因损害关键因素而导致正常眼压性青光眼。 我的建议旨在：（i）确定功能。 RPAP3编码变异对HSP90相互作用/刺激和ECM基因表达的影响， (ii) 确定诊断为青光眼的患者队列中有害 RPAP3 变异的患病率， (iii) 阐明在这个大家族中分离的其他青光眼风险变异。结果可能导致。 通过实现这些目标，我还将改善 NTG 患者的筛查和治疗选择。 拓宽我在疾病基因发现和遗传变异功能表征方面的技能，让我做好准备 专注于人类疾病遗传学和分子诊断的职业。