Human embryonic stem cell derived midbrain dopamine neurons

描述(申请人提供)：该提案的目标是在帕金森病(PD)的临床前模型中确定限制人类ES(HES)来源的多巴胺(DA)神经元在体内存活和功能的因素。我们实验室的初步工作为从HES细胞衍生出中脑DA神经元提供了高产量的方案。然而，在啮齿动物和非人类灵长类动物体内进行的HES细胞来源的DA神经元的试验性移植研究表明，DA在体内的存活率和维系能力很差。体内存活率较低与使用小鼠胚胎来源的DA神经元形成对比，后者在同种和异种移植帕金森病模型中都显示出良好的存活率。这表明HES细胞来源的DA神经元表现出一种特殊的脆弱性，在这些细胞的临床应用得以设想之前，需要解决这种脆弱性。在此，我们建议系统地探讨在帕金森病的啮齿动物和猴子模型中，细胞或宿主的参数或两者的组合是否对体内有限的DA神经元存活和功能负责。这些研究为我们长期致力于将基于HES细胞的DA神经元分化策略转化为临床应用奠定了基础。这项研究有两个主要目的： 1.探讨体内存活的DA神经元数量减少是否与体外中脑DA神经元分化不完全有关 1.1.改善HES来源的神经花环对中脑DA神经元的识别 1.2.神经花环形成的时机：作为DA神经元来源的早期花环与晚期花环。 2.探讨体内存活的DA神经元数量减少是否与移植的HES来源的DA神经元群体选择性死亡有关。 2.1.HES来源的DA神经元的固有脆弱性 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