Human Cell and Gene Therapy in Parkinsonian monkeys

描述（由申请人提供）：帕金森氏症是一种毁灭性的、普遍致命的、无法治愈的神经退行性疾病，它使人衰弱，并对患者的身心造成可怕的痛苦。人类多能细胞，包括胚胎干细胞，已被确定为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损伤的猴子）中测试长期治疗效果。我们的长期目标是评估HESC-DA细胞治疗PD患者的安全性和短期有效性。目前的目标是确定移植HESC-DA细胞是否是一种有效的持久治疗方法，可以在多巴胺缺失的灵长类大脑中进行结构整合，并逆转PD相关的功能缺陷。我们的中心假设是，脑膜内输送HESC-DA细胞将逆转MPTP治疗的猴子的运动障碍，这种效果将通过与AAV-neurturin共同治疗而增强。这项研究的基本原理是，如果HESC-DA移植可以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合乎逻辑的步骤。在初步数据的指导下，该假设将验证以下特定目标，即在MPTP猴子的纹状体内移植HESC-DA细胞，或HESC-DA细胞+ AAV2-Neurturin将提供结构和功能恢复。利用一种新颖的基于底板的多巴胺能神经化模式，使HESC能够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）联系起来。在拟议的目标中，免疫组织化学分析和行为测试将用于补充体内成像，以弥合PD治疗知识的关键空白。这项研究具有创新性，因为它1)专注于特征明确、功能活跃的中脑特异性HESC- DA神经元，作为细胞移植的可再生替代来源；2)利用临床相关的非人灵长类动物PD模型，提高了我们对该方法功能功效的认识；3)通过增加AAV-2神经营养支持，垂直推进和连接了我们目前的研究。这一建议具有重要意义，因为在帕金森猴子中严格测试HESC-DA疗法更准确地描述了人类PD，因此，提议的实验更有可能为临床提供高质量的可翻译结果。这一结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。在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患者的整体生活质量。

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.