利用三维多层水凝胶系统研究Rett综合症中星型胶质细胞的作用及其机制

三维培养系统更加接近在体细胞的空间环境，能够提供更加符合真实生理情况的细胞生长外环境：组织特异性结构、机械和多细胞相互作用等条件，因此更加适合于对复杂的神经系统内各种细胞的分化以及迁移的研究。本课题试图建立适合神经前体细胞向星型胶质细胞分化的三维多层水凝胶系统，以期解决二维培养系统无法快速、稳定、高效的进行星型胶质细胞分化的问题，并将其应用与RTT疾病中星型胶质细胞可能存在的功能缺陷的研究中。采用三维水凝胶方法，我们将对RTT病人iPSCs诱导分化的神经前体细胞向星型胶质细胞的分化能力、分化的星型胶质细胞诱导神经前体细胞迁移以及对神经元分化和成熟的调控能力予以评估，试图阐明RTT病人星型胶质细胞与正常细胞的差异及其分子机制，为RTT疾病的发病机制提供新的思路。进而我们将以三维系统为模型，筛选能够恢复RTT病人星型胶质细胞功能的小分子化合物，以期为RTT疾病的治疗提供具有临床应用价值的药物。

Probing a wide range of cellular phenotypes in neurodevelopmental disorders using patient-derived neural progenitor cells (NPCs) can be facilitated by three-dimensional (3D) assays, as 3D systems enable the formation of tissue-mimetic architectures and promote more realistic physiological responses of brain tissue than conventional 2D. Take consideration of the important function of astrocytes in neural maturation and homeostasis, we plan to optimize a 3D layered hydrogel for better astrocyte differentiation from iPSCs-derived NPCs. Through comparing of the migration and maturation of NPCs induced by astrocyte from either Rett syndrome (RTT) patients or normal persons, we will try to identify a MeCP2 genotype-specific defect of astrocyte in RTT patients and reveal the molecular mechanism for this defect. Further，using our novel layered 3D culture system, we will set up a high-throughput drug-screening platform targeting the defect of astrocytes for RTT disease. Consequently, our reliable 3D based differentiation methods, which can potentially enrich and accelerate astrocyte differentiation and neuron maturation and migration, would be beneficial in order to improve our understanding of RTT pathology and to discover new pharmaceutical drug for the treatment of RTT.