Human Cell and Gene Therapy in Parkinsonian monkeys

描述（由申请人提供）：帕金森病是一种毁灭性的、普遍致命的、无法治愈的神经退行性疾病，它会使人衰弱，并给患者的身心带来可怕的痛苦。人类多能细胞，包括胚胎干细胞，已被鉴定为用于治疗的潜在替代细胞基质，但迄今为止，尚未在疾病相关动物模型（MPTP损伤的猴）中测试长期治疗功效。我们的长期目标是评估HESC-DA细胞疗法在PD患者中的安全性和暂时有效性。目前的目标是确定移植的HESC-DA细胞是否是一种有效的持久疗法，其在结构上整合在多巴胺耗尽的灵长类动物大脑中，以及逆转与PD相关的功能缺陷。我们的中心假设是HESC-DA细胞的纹状体内递送将逆转MPTP治疗的猴中的运动失能，并且这种作用将通过与AAV-neurturin共治疗而增强。拟议研究的基本原理是，如果HESC-DA移植物改善帕金森病灵长类动物的功能缺陷，我们可以进行大规模，长期的安全性和耐受性研究，作为临床转化的下一个逻辑步骤。在初步数据的指导下，该假设将测试以下特定目的，即HESC-DA细胞或HESC-DA细胞+AAV 2-Neurturin的纹状体内移植将在MPTP猴中提供结构和功能恢复。利用一种新的基于底板的多巴胺能神经化范例，使HESC有效地分化为中脑特异性多巴胺能神经元，我们将确定移植的HESC-DA细胞对宿主解剖结构和功能的影响，并将这些结果与整体生活质量（临床评分量表）相关联。在提出的目标中，免疫组织化学分析和行为测试将用于补充体内成像，以弥补PD治疗知识的关键空白。该研究是创新性的，因为它1）集中于充分表征的、功能活性的、中脑特异性HESC-DA神经元作为细胞移植的替代可再生来源2）通过利用临床相关的非人灵长类PD模型来推进我们对该方法的功能功效的认识，以及3）通过添加AAV-2 neurturin神经营养支持来纵向推进和桥接我们目前的研究。这一建议是重要的，因为在帕金森病猴中严格测试HESC-DA疗法更准确地描述了人类PD，因此，所提出的实验更有可能为临床提供高质量的可翻译结果。这些成果有望通过神经外科和干细胞生物学的融合垂直推进PD治疗领域。这里获得的知识有可能提供一种治疗选择，将减少可怕的症状，提高PD的整体生活质量。

DESCRIPTION (provided by applicant): Parkinson's disease is a devastating, universally fatal, incurable neurodegenerative disorder, which is debilitating and inflicts terrible suffering on the patient's mind and body. Human pluripotent cells, including embryonic stem cells, have been identified as a potential alternative cellular substrate for treatment, but to date, have not been tested for long-term therapeutic efficacy in disease relevant animal models (MPTP lesioned monkeys). Our long-term goal is to assess the safety and temporal efficacy of HESC-DA cell therapy in PD patients. The current objective is to determine if grafted HESC-DA cells are a potent long-lasting therapy that structurally integrate within the dopamine depleted primate brain, as well as reverse functional deficits associated with PD. Our central hypothesis is that intrastriatal delivery of HESC-DA cells will reverse motor disability in MPTP treated monkeys and this effect will be potentiated by co-treating with AAV-neurturin. The rationale for the proposed research is if HESC-DA grafts ameliorate functional deficits in parkinsonian primates, we can pursue large-scale, long-term safety and tolerability studies as the next logical step for clinical translation. Guided by preliminary data, this hypothesis will test the following Specific Aim, that intrastriatal grafting of HESC-DA cells, or HESC-DA cells + AAV2-Neurturin will provide structural and functional recovery in MPTP monkeys. Utilizing a novel floor-plate based, dopaminergic neuralization paradigm that allows HESC to be efficiently differentiated into midbrain specific dopaminergic neurons, and we will determine the effects of grafted HESC-DA cells on host anatomical structure and function and correlate these results to overall quality of life (clinical rating scale). In the proposed Aim, immunohistochemical analysis and behavioral testing will be used to complement in vivo imaging in an effort to bridge a critical gap in knowledge for PD therapeutics. This research is innovative, as it 1) focuses on well characterized, functionally active, midbrain specific HESC- DA neurons as an alternative renewable source for cellular transplantation 2) advances our knowledge of the functional efficacy of this approach by utilizing a clinically relevant non-human primate model of PD, and 3) vertically advances and bridges our present studies with the addition of AAV-2 neurturin neurotrophic support. This proposal is significant as rigorously testing HESC-DA therapies in parkinsonian monkeys more accurately depicts human PD, and therefore, the proposed experiments are more likely to provide quality translatable results to the clinic. The outcomes are expected to vertically advance the field of PD therapy through the blending of neurosurgery and stem cell biology. The knowledge obtained here has the potential to provide a therapeutic option that will reduce the terrible symptoms and enhance overall quality of life in PD.