Human organoid modelling of neurogenetic epilepsy syndromes

神经源性癫痫综合征的人体类器官模型

• 批准号: 2645811
• 金额: --
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2645811
• 关键词: Human; organoid; modelling; neurogenetic; epilepsy

Rare single gene disorders manifest in the neonatal period. The Next Generation Children's Project has used whole genome sequencing of patients and their parents to diagnose 50 patients with neurogenetic epilepsy syndromes. For example, Kv7.2 voltage-gated potassium channel subunit dysfunction is associated with KCNQ2 mutations, with resulting increased network excitability. It is increasingly clear that epilepsy-causing gene mutations affect the major glial cell populations, principally astrocytes, rather than neurons. To date, human induced pluripotent stem cell-derived (iPSC) neurons have been generated with epilepsy-causing potassium/sodium channel mutations and gene deletions, however, lack the ability to establish human cortical connections. The development of human three-dimensional cerebral organoid models (CO) that incorporate circuit forming neuron and glial phenotypes present a unique opportunity to develop new approaches in precision medicine for paediatric epilepsy patients. This project aims to generate CO from CRISPR/Cas9 KCNQ2 and KCNJ10-edited H9 ECS lines and from patient-derived iPSC-line. To assess if stem cell-derived CO harbouring KCNQ2 or KCNJ10 mutations show altered cortical plate development, assessing CO cell composition by immunolabeling for cell type-specific markers (GAD2/ CTIP2+). To show whether CO slices with perturbed KCNQ2 and KCNJ10 function or expression show bursts in activity and whether this can be suppressed by Na+ or K+ channel drugs.

罕见的单基因疾病表现在新生儿期。下一代儿童项目使用患者及其父母的全基因组测序来诊断50名神经源性癫痫综合征患者。例如，Kv7.2电压门控钾通道亚基功能障碍与KCNQ 2突变相关，导致网络兴奋性增加。越来越清楚的是，致癫痫基因突变影响主要的神经胶质细胞群，主要是星形胶质细胞，而不是神经元。迄今为止，人类诱导多能干细胞衍生（iPSC）神经元已经产生了癫痫引起的钾/钠通道突变和基因缺失，然而，缺乏建立人类皮层连接的能力。结合电路形成神经元和神经胶质表型的人类三维脑类器官模型（CO）的开发为儿科癫痫患者的精准医学开发新方法提供了独特的机会。该项目旨在从CRISPR/Cas9 KCNQ 2和KCNJ 10编辑的H9 ECS系以及从患者来源的iPSC系产生CO。通过免疫标记细胞类型特异性标志物（GAD 2/CTIP 2+）评估CO细胞组成，评估携带KCNQ 2或KCNJ 10突变的干细胞源性CO是否显示皮质板发育改变。显示KCNQ 2和KCNJ 10功能或表达受干扰的CO切片是否显示活性爆发，以及这是否可以被Na+或K+通道药物抑制。