Modeling of early-onset retinal dystrophy development in optic vesicle containing-brain organoids

含有脑类器官的视泡中早发性视网膜营养不良发育的建模

• 批准号: 399443882
• 负责人: Professor Dr. Jay Gopalakrishnan, Ph.D.
• 金额: --
• 依托单位: Institut für Molekulare Physiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/399443882?language=en
• 关键词: Modeling; early; onset; retinal; dystrophy

Congenital retinal disorders, including Leber Congenital Amaurosis (LCA) /early-onset severe retinal dystrophy and Oculocutaneous Albinism (OCA), are genetically inherited retinal disorders affecting ~150 live births every year. These congenital retinal disorders occur at the early onset of retinal development, for which the mechanisms of disease development remain unknown. Therefore, underpinning the defective cellular mechanisms that trigger the disease development in a disease-relevant experimental system is a prerequisite to designing therapeutic strategies. A significant limiting factor for this is the lack of an experimental system that physiologically recapitulates the early retinogenesis from the developing forebrain. The first funding period of this priority prograhas supported us to generate 3D brain organoids developing bilaterally symmetric optic vesicles in a topographically restricted manner (OVB-organoids). The OVB-organoid culturing method is robust as we could generate numerous OVB-organoids from five independent iPSCs donors. In the current funding period, we will model the developmental disease mechanisms of LCA and OCA using patient-specific OVB-organoids and test if pharmacological agents and gene augmentation strategies can mitigate the disease phenotypes.To this end, via collaborating with human geneticists, we have identified patients carrying mutations in SPATA7, Cep290 (causing LCA), and HPS1 (causing OCA). Notably, these patient mutations are non-syndromic, meaning that the mutations do not affect the brain developmental process but only the retina.This research program combines our interdisciplinary expertise to dissect the deregulated cellular pathways that elicit disease development in patient-specific OVB-organoids and evaluate potential gene-based therapies. To accomplish this ultimate goal, we will1. Generate vascularized OVB-organoids (v-OVB-organoids) for an extended period of viability to generate mature retinal cell types.2. Generate patient-specific v-OVB-organoids to dissect developmental pathophysiology of LCA and OCA due to SPATA7, Cep290, and HPS1 mutations.3. Determine the impact of translational read-through agents and AAV-gene augmentations in mitigating the disease phenotypes. Overall, with our interdisciplinary expertise that uses novel types of 3D brain-retinal hybrid organoids, we will identify deregulated cellular pathways of retinal developmental disorders causing LCA and OCA and offer novel therapeutic options.

先天性视网膜疾病，包括Leber先天性黑蒙（LCA）/早发性严重视网膜营养不良和眼皮肤白化病（OCA），是遗传性视网膜疾病，每年影响约150例活产婴儿。这些先天性视网膜疾病发生在视网膜发育的早期，其疾病发展的机制仍然未知。因此，在疾病相关的实验系统中支持引发疾病发展的缺陷细胞机制是设计治疗策略的先决条件。一个重要的限制因素是缺乏一个实验系统，生理重演早期视网膜发生从发展中的前脑。该优先级的第一个资助期支持我们生成3D脑类器官，以地形限制的方式开发双侧对称的视囊泡（OVB类器官）。OVB-类器官培养方法是稳健的，因为我们可以从五个独立的iPSC供体产生许多OVB-类器官。在当前的资助期内，我们将使用患者特异性OVB类器官来模拟LCA和OCA的发育疾病机制，并测试药物和基因增强策略是否可以减轻疾病表型。为此，我们通过与人类遗传学家合作，确定了携带SPATA 7，Cep290（导致LCA）和HPS1（导致OCA）突变的患者。值得注意的是，这些患者突变是非综合征性的，这意味着这些突变不会影响大脑发育过程，而只会影响视网膜。这项研究计划结合了我们的跨学科专业知识，剖析了在患者特异性OVB类器官中引起疾病发展的失调细胞通路，并评估了潜在的基因治疗方法。为了实现这一最终目标，我们将1。 产生血管化的OVB-类器官（v-OVB-类器官），持续延长的存活期，以产生成熟的视网膜细胞类型。 生成患者特异性v-OVB类器官以剖析由于SPATA 7、Cep290和HPS1突变导致的LCA和OCA的发育病理生理学。 确定翻译通读剂和AAV基因扩增对减轻疾病表型的影响。总的来说，凭借我们使用新型3D脑-视网膜混合类器官的跨学科专业知识，我们将确定导致LCA和OCA的视网膜发育障碍的失调细胞通路，并提供新的治疗选择。