Generation of Cortical Motoneurons from Embryonic Stem Cells

从胚胎干细胞产生皮质运动神经元

• 批准号: 7582618
• 负责人: XUE-JUN LI
• 金额: $ 12万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7582618
• 关键词: Amyotrophic Lateral Sclerosis; Brain Stem; Cell Therapy; Cells; Cerebral cortex; Chick Embryo; Disease; Dorsal; Embryo; Environment; Fibroblast Growth Factor; Future; Generations; Human; In Vitro; Inherited; Motor Neurons; Movement; Muscle; Neuroepithelial; Neuroepithelial Cells; Neurons; Pharmacologic Substance; Primary Lateral Sclerosis; Process; Prosencephalon; Registries; Role; Screening procedure; Signal Transduction; Source; Spastic; Spastic Hemiplegias; Specific qualifier value; Spinal; Spinal Cord; Staging; System; Telencephalon; Testing; Transplantation; United States National Institutes of Health; WA01 cell line; WA09 Cell Line; base; embryonic stem cell; human embryonic stem cell; in vivo; morphogens; nervous system disorder; neurotrophic factor; progenitor; relating to nervous system; success; tool

DESCRIPTION (provided by applicant): motoneurons in the cerebral cortex, also known as upper motoneurons, control our muscle movement through lower motoneurons in the brain stem and spinal cord. Degeneration of upper motoneurons results in progressive spastic and weakness in muscles, which underlies some debilitating neurological disorders, such as amyotrophic lateral sclerosis (ALS), hereditary spastic hemiplegia and primary lateral sclerosis. There is no effective treatment for these motoneuron disorders, for the most part due to the lack of understanding how cortical motoneurons are born, functionally mature and degenerate. This application, based on our success in producing spinal motoneurons from human embryonic stem cells (ESCs), will explore the feasibility of generating upper motoneurons from self-renewing human ESCs (H1 and H9 lines, NIH Registry WA01 and WA09). We have established a chemically defined culture system to direct human ESCs to neuroepithelial cells and discovered a critical primitive neuroepithelial stage for specification of region-specific neuronal subtypes. We will first induce differentiation of dorsal telencephalic neural progenitors by examining the effect of such morphogens as Wnts, Wnt antagonists, and fibroblast growth factors (FGFs), individually, in combination, or sequentially. These progenitors will then be further differentiated to motoneurons in the presence of neurotrophic factors and/or target cells (lower motoneuron) and/or local environment. The function of in vitro generated upper motoneurons will be assessed by their interaction with lower motoneurons in culture and following transplantation to dorsal telencephalon of a chick embryo. This application will thus produce the currently unavailable upper motoneurons from limitless ESCs and offer an otherwise inaccessible tool for studying how human upper motoneurons are born, functionally mature, and become sick. These cells will also provide a target for screening pharmaceuticals that stop the process of motoneuron degeneration and a source for potential future cell therapy for upper motoneuron related diseases.

描述（由申请人提供）：大脑皮层中的运动神经元，也称为上运动神经元，通过脑干和脊髓中的下运动神经元控制我们的肌肉运动。上运动神经元的退化导致肌肉进行性痉挛和无力，这是一些使人衰弱的神经系统疾病的基础，例如肌萎缩侧索硬化症（ALS）、遗传性痉挛性偏瘫和原发性侧索硬化症。这些运动神经元疾病没有有效的治疗方法，很大程度上是由于缺乏对皮质运动神经元如何产生、功能成熟和退化的了解。该应用基于我们从人类胚胎干细胞（ESC）产生脊髓运动神经元的成功，将探索从自我更新的人类ESC（H1和H9系，NIH登记号WA01和WA09）产生上运动神经元的可行性。我们建立了化学成分确定的培养系统，将人类 ESC 引导至神经上皮细胞，并发现了用于指定区域特异性神经元亚型的关键原始神经上皮阶段。我们将首先通过单独、组合或顺序检查诸如 Wnt、Wnt 拮抗剂和成纤维细胞生长因子 (FGF) 等形态发生素的作用来诱导背侧端脑神经祖细胞的分化。然后，在神经营养因子和/或靶细胞（下运动神经元）和/或局部环境存在的情况下，这些祖细胞将进一步分化为运动神经元。体外产生的上运动神经元的功能将通过它们与培养物中的下运动神经元的相互作用以及移植到鸡胚胎的背侧端脑后进行评估。因此，该应用程序将从无限的 ESC 中产生目前无法获得的上运动神经元，并为研究人类上运动神经元如何出生、功能成熟和生病提供一种其他方式无法获得的工具。这些细胞还将为筛选阻止运动神经元变性过程的药物提供靶标，并为未来上运动神经元相关疾病的潜在细胞疗法提供来源。