Human Disease Modeling of Glaucomatous Neuropathy

青光眼神经病的人类疾病模型

• 批准号: 10357852
• 负责人: Iqbal Ahmad
• 金额: $ 36.98万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10357852
• 关键词: Abnormal Cell; Adult; Affect; Age; Animal Model; Axotomy; Blindness; Blood specimen; Cell Differentiation process; Cells; Chemicals; Clinical; Collaborations; Complex; DNA Binding; Data; Data Analyses; Development; Diagnosis; Discrimination; Disease; Disease model; Early Diagnosis; Embryo; FRAP1 gene; Generations; Genes; Genetic Transcription; Genetic Variation; Glaucoma; HMGA2 gene; Human; Knowledge; Letters; Light; Microfluidic Microchips; Microfluidics; Missense Mutation; Modeling; Molecular; Natural regeneration; Neuropathy; Nucleotides; Ocular Hypertension; Onset of illness; Optic Nerve; Pathway Analysis; Pathway interactions; Patients; Phenotype; Physiologic Intraocular Pressure; Predisposition; Primary Open Angle Glaucoma; Process; Progressive Disease; Protocols documentation; Regulator Genes; Research; Retina; Retinal Ganglion Cells; Risk Factors; Signal Transduction; Technology; Testing; Therapeutic; Thick; Transplantation; Variant; Visual Fields; Zebrafish; axon growth; axon regeneration; base; blind; cellular pathology; clinically significant; early detection biomarkers; experimental study; genetic variant; genome-wide; high intraocular pressure; homeodomain; human disease; in vivo; induced pluripotent stem cell; regeneration potential; regenerative; retinal ganglion cell regeneration; retinal nerve fiber layer; retinal progenitor cell; risk variant; sex; stem cell model

PROJECT SUMMARY Glaucoma represents a group of diseases, which are associated with multiple risk factors and genetic variants. The unifying theme among these diseases is the progressive degeneration of optic nerve and retinal ganglion cells (RGCs) degenerate, leading to irreversible blindness. Based on this observation we propose a hypothesis that RGCs are intrinsically vulnerable to glaucoma risk factors and genetic variants. Since RGCs are born embryonically and glaucoma is an adult onset disease our knowledge about the emergence of and mechanism underlying RGC susceptibility, important for early diagnosis and formulating therapeutic approaches, remain rudimentary. Our objective is to examine the impact of genetic variations associated with glaucoma on the development, phenotype, and regeneration of RGCs, using the induced pluripotent stem cell (iPSC) model of primary open angle glaucoma (POAG), the most prevalent type of the disease. Here, we will test the hypothesis that RGCs in the SIX6 risk allele (Asn141His, rs33912345) in POAG are developmentally compromised in normal phenotype and function, and that this affects their survival and regeneration. The SIX6 variant is a suitable target for the analysis because SIX6 is one of the eye-field genes involved in retinal development and the missense mutation (Asn141His) in the DNA binding region of SIX6 is accompanied by degenerative changes that include reduction in the retinal nerve fiber layer (RNFL) thickness in POAG. Thus, the generation of SIX6 risk allele-POAG RGCs in a dish model of the disease would allow the characterization of developmental and phenotype abnormalities, shedding light on glaucomatous RGC susceptibility. We have proposed three aims to test the central hypothesis. First, we will determine the impact of the SIX6 risk allele-POAG on the development and phenotype of patient-specific RGCs by systematic characterization of sequential steps of retinal inductions and RGC differentiation of POAG patient-specific and age- and sex-matched control iPSCs, under the influence of a stage-specific and chemically defined protocol. Second, we will examine the impact of the SIX6 risk allele-POAG on patient-specific RGC axon growth and regeneration in the context of mTOR signaling, a regulator of retinal development and regeneration, which is inhibited in POAG patient- specific RGCs. Lastly, we will identify the molecular pathway(s) underlying abnormal RGC development and phenotype. We will carry out hypothesis-driven (e.g., HMGA2 and KLF-4 as candidate hub genes) genome wide transcriptional analysis of POAG patient-specific RGCs during development and regeneration to understand the molecular mechanism underlying SIX6 risk allele associated RGC abnormalities. Information emerging from our study will bridge a gap in our knowledge of the intrinsic vulnerability of RGCs in glaucoma. The information on dysregulated processes and pathways will reveal biomarkers for early diagnosis, and will allow strategies for therapeutic regeneration of glaucomatous RGCs.

项目摘要 青光眼是一组与多种危险因素和遗传变异相关的疾病。的 这些疾病的共同主题是视神经和视网膜神经节细胞（RGC）的进行性变性 退化，导致不可逆转的失明。基于这一观察，我们提出了一个假设，即RGC是 本质上易受青光眼危险因素和遗传变异的影响。由于视网膜神经节细胞在胚胎中出生， 是一种成人发病的疾病，我们对RGC易感性的出现和机制的了解， 对于早期诊断和制定治疗方法，仍然是基本的。我们的目标是研究 与青光眼相关的遗传变异对RGCs发育、表型和再生的影响， 本发明涉及一种用于治疗原发性开角型青光眼（POAG）的多能干细胞（iPSC）模型，所述原发性开角型青光眼是最普遍的疾病类型。在这里， 我们将检验POAG中SIX 6风险等位基因（Asn 141 His，rs33912345）中的RGC在发育上与POAG中的RGC相似的假设。 在正常表型和功能中受损，并且这影响它们的存活和再生。SIX 6变体是一种 因为SIX 6是参与视网膜发育的眼区基因之一， SIX 6 DNA结合区的错义突变（Asn 141 His）伴随着退行性变化，包括 POAG中视网膜神经纤维层（RNFL）厚度的减少。因此，SIX 6风险等位基因-POAG的产生 该疾病的培养皿模型中的RGC将允许表征发育和表型异常， 揭示了视网膜节细胞癌的易感性。我们提出了三个目标来检验中心假设。一是 将确定SIX 6风险等位基因-POAG对患者特异性RGC的发育和表型的影响， 视网膜诱导的连续步骤的系统表征和POAG患者特异性的RGC分化 年龄和性别匹配的对照iPSC，在阶段特异性和化学定义的方案的影响下。二是 将研究SIX 6风险等位基因POAG对患者特异性RGC轴突生长和再生的影响， mTOR信号传导的背景，视网膜发育和再生的调节剂，在POAG患者中被抑制- 具体的RGC。最后，我们将确定异常RGC发育和表型的分子途径。 我们将进行假设驱动（例如，HMGA 2和KLF-4作为候选枢纽基因）全基因组转录 分析POAG患者特异性RGCs在发育和再生过程中的分子机制 潜在的SIX 6风险等位基因相关的RGC异常。从我们的研究中产生的信息将弥合我们之间的差距， 了解青光眼中RGCs的内在脆弱性。关于失调过程和途径的信息 将揭示用于早期诊断的生物标志物，并将为青光眼RGC的治疗性再生提供策略。