Human embryonic stem cell derived midbrain dopamine neurons

描述（由申请人提供）：该提案的目标是确定限制帕金森病（PD）临床前模型中人ES（hES）衍生的多巴胺（DA）神经元体内存活和功能的因素。我们实验室的初步工作为从 hES 细胞中衍生出中脑 DA 神经元提供了高产方案。然而，对啮齿动物和非人灵长类动物的 hES 细胞来源的 DA 神经元进行的初步移植研究表明，DA 命运的体内存活和维持较差。体内存活率较低，与小鼠 ES 衍生的 DA 神经元的研究形成鲜明对比，后者在同种异体和异种移植 PD 模型中均表现出强劲的存活率。这表明 hES 细胞衍生的 DA 神经元表现出一种特殊的脆弱性，在设想这些细胞的临床应用之前需要解决这一脆弱性。在这里，我们建议系统地解决基于细胞或宿主的参数或两者的组合是否对 PD 啮齿动物和猴子模型中有限的体内 DA 神经元存活和功能负责。这些研究应该为我们长期努力将基于 hES 细胞的 DA 神经元分化策略转化为临床应用奠定基础。该研究有两个主要目标： 1. 探讨体内存活的 DA 神经元数量低是否是由于体外中脑 DA 神经元分化不完全所致 1.1.改善 hES 衍生的神经玫瑰花结的模式，以实现中脑 DA 神经元身份 1.2.神经玫瑰花结模式形成的时间：早期与晚期玫瑰花结作为 DA 神经元的来源。 2. 探讨体内存活 DA 神经元数量低是否是由于移植的 hES 衍生 DA 神经元群的选择性细胞死亡所致。 2.1. hES 衍生的 DA 神经元的内在脆弱性 2.2.体内缺乏足够的营养支持 2.3.宿主免疫/炎症反应干扰 DA 神经元存活

DESCRIPTION (provided by applicant): The goal of the proposal is to identify the factors that limit in vivo survival and function of human ES (hES) derived dopamine (DA) neurons in preclinical models of Parkinson's disease (PD). Preliminary work in our lab has provided high-yield protocols for the derivation of midbrain DA neurons from hES cells. However, pilot transplantation studies of hES cell derived DA neurons in rodent and non-human primates suggest poor in vivo survival and maintenance of DA fate. Poor in vivo survival is in contrast to work with mouse ES derived DA neurons that showed robust survival in both allo- and xenograft PD models. This suggests that hES cell derived DA neurons exhibit a particular vulnerability that needs to be resolved before the clinical use of these cells can be envisaged. Here we propose to systematically address whether cell or host-based parameters or a combination of the two are responsible for the limited in vivo DA neuron survival and function in rodent and monkey models of PD. These studies should provide the basic foundation for our long-term efforts to translate hES cell based DA neuron differentiation strategies for clinical application. The Study has two major aims: 1. To address whether low numbers of surviving DA neurons in vivo are due to incomplete midbrain DA neuron differentiation in vitro 1.1. Improving patterning of hES derived neural rosettes towards midbrain DA neuron identity 1.2. Timing of patterning in neural rosettes: Early vs. late rosettes as a source of DA neurons. 2. To address whether low numbers of surviving DA neurons in vivo are due to selective cell death of the grafted hES-derived DA neuron population. 2.1. Intrinsic vulnerability of hES derived DA neurons 2.2. Lack of sufficient trophic support in vivo 2.3. Host immunological/inflammatory response interferes with DA neuron survival