Nanofibrous Scaffolds for Transplantation of Human Dopaminergic Neurons

 描述(申请人提供)：这个项目旨在设计创新的支架，将综合解决治疗神经退行性疾病的两个关键障碍：(A)细胞来源：支持体外重新编程的人类干细胞来源的神经元的成熟、规格和功能，以及(B)特定亚型的神经元移植：能够有效地移植来治疗体内的神经退行性疾病。核心假设是基于纳米纤维水凝胶支架的三维工程微生态位(EMN)可以在移植前支持亚型特定神经元的体外诱导和成熟，并促进移植后的存活和增强与宿主组织的功能相互作用。对该项目感兴趣的一个具体应用是治疗神经退行性疾病，如帕金森氏病(PD)。为了实现我们的目标，我们提出了两个具体目标。第一个目标是设计诱导多能干细胞(IPSC)来源的重编程多巴胺能(DA)神经元的成熟引导EMN。三维EMN将以转录因子转导的IPSCs为基础，培养在纳米纤维水凝胶中，这些水凝胶是由自组装的最低限度免疫原肽制成的。为了指导DA神经元的成熟和规范，将使用亚型特定线索对EMN进行功能化。我们将确定移植细胞的新亚群，并通过全基因组测序检查神经支配的宿主组织的变化。第二个目标是将DA和兴奋性神经元的自激EMN移植到小鼠PD模型的纹状体中。我们假设，在微支架环境中，一个自我运作的E-DA微型神经回路将提供足够的兴奋性驱动，以促进体内DA神经元与宿主组织功能增强的相互作用。我们将把E和DA神经元的自激EMN移植到缺乏DA神经支配的小鼠的纹状体内。我们将评估E和DA神经元的自激EMN改善帕金森病症状中的功能缺陷的能力，并再次检查移植细胞和神经支配的宿主组织的成熟结果。这项研究的总体结果将有助于解决神经退行性疾病细胞替代疗法领域中的关键障碍，如移植组织的功能和存活。

 DESCRIPTION (provided by applicant): This project aims to design innovative scaffolds that will integratively address two critical barriers for treating neurodegenerative diseases: (a) Cell Sourcing: support the maturation, specification, and function of reprogrammed human stem cell-derived neurons in vitro and (b) Subtype-specific Neuronal Transplantation: enable efficacious transplantation to treat neurodegenerative diseases in vivo. The central hypothesis is that 3D engineered microscale niches (EMNs) based on nanofibrous hydrogel scaffolds can support the induction and maturation of subtype specific neurons in vitro prior to transplantation and promote the survival and enhanced functional interaction with host tissue following transplantation. A specific application of interest to this project is the treatment of neurodegenerative diseases like Parkinson's disease (PD). To achieve our goal, two specific aims are proposed. The first aim is concerned with designing maturation-guiding EMNs of induced pluripotent stem cell (iPSC)-derived reprogrammed dopaminergic (DA) neurons. The 3-D EMNs will be based on transcription factor-transduced iPSCs cultured within nanofibrous hydrogels fabricated from self-assembling minimally immunogenic peptides. To guide the maturation and specification of the DA neurons, the EMNs will be functionalized with subtype specific cues. We will determine the emergent subpopulations of transplanted cells and examine changes in innervated host tissue through whole genome sequencing. The second aim will be focused on transplanting self-actuating EMNs of DA and excitatory neurons into the striatum of a mouse PD model. We hypothesize that a self-functioning E-DA mini neural circuitry within a microscaffold environment will provide sufficient excitatory drive to promote enhanced functional interaction of DA neurons with host tissue in vivo. We will transplant the self-actuating EMNs of E and DA neurons into the striatum of mice lacking DA innervations. We will evaluate the ability of self-actuating EMNs of E and DA neurons to improve functional deficits in the Parkinson's disease symptoms, and again examine maturation outcomes of transplanted cells and innervated host tissues. The overall outcomes from this study will help to address the critical barriers such as the functioning and survival of transplanted tissue in the field of cell-replacement therapies for neurodegenerative diseases.