Muscle stem cells: New ALS growth factor therapy and disease model

肌肉干细胞：新的 ALS 生长因子疗法和疾病模型

• 批准号: 9002105
• 负责人: Masatoshi Suzuki
• 金额: $ 33.47万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9002105
• 关键词: Address; Adult; Amyotrophic Lateral Sclerosis; Biology; Cause of Death; Cell Line; Cell Survival; Cell Therapy; Cell model; Cells; Cessation of life; Characteristics; Coculture Techniques; Cultured Cells; Data; Disease; Disease Progression; Disease model; Environment; Equus caballus; Failure; Familial Amyotrophic Lateral Sclerosis; Future; Gene Mutation; Goals; Growth; Health; Heterogeneity; Human; Implant; In Vitro; Knowledge; Laboratories; Limb structure; Longevity; Methods; Modeling; Motor; Motor Endplate; Motor Neurons; Motor output; Muscle; Muscle Cells; Muscle Fibers; Muscle satellite cell; Muscular Atrophy; Neurodegenerative Disorders; Neuromuscular Diseases; Neuromuscular Junction; Neurosciences; Outcome; Paralysed; Patients; Play; Process; Progressive Disease; Publishing; Rattus; Research; Respiratory Diaphragm; Respiratory Failure; Respiratory Paralysis; Rodent Model; Role; Skeletal Muscle; Societies; Stem cell transplant; Stem cells; Testing; Therapeutic; Tissues; Translations; Transplantation; United States National Institutes of Health; Vascular Endothelial Growth Factors; Work; base; cell growth; effective therapy; glial cell-line derived neurotrophic factor; human disease; improved; in vitro Model; induced pluripotent stem cell; insight; motor neuron degeneration; muscle regeneration; neuromuscular; novel; preclinical study; progenitor; repaired; respiratory; stem cell biology; stem cell therapy; treatment strategy

 DESCRIPTION (provided by applicant): Amyotrophic lateral sclerosis (ALS) is a progressive disease causing motor neuron degeneration, muscular atrophy and, ultimately, death by respiratory failure. Our major goal in this project is to determine if newly established human skeletal muscle progenitor/stem cells (hSMPCs) derived from induced pluripotent stem cells (iPSCs) can be used for ex vivo cell therapy (stem cell-based growth factor delivery), and as an in vitro model to study ALS. The fundamental hypothesis guiding this proposal is that iPSC-derived hSMPCs efficiently differentiate into new skeletal muscle cells and contribute to muscle regeneration. This capacity confers the capacity for iPSC-derived hSMPCs to deliver ex vivo growth factors, and to model aspects of ALS in vitro. Our hypothesis is based on our published works and new preliminary data demonstrating the feasibility of producing hSMPCs from iPSCs. We will prepare genetically modified hSMPCs to deliver key growth factors known to be neuroprotective in ALS rodent models, including glial cell line-derived neurotrophic factor (GDNF) and vascular endothelial growth factor (VEGF). After establishing the cells, we will transplant them into the limb muscles to deliver growth factors in ALS rats (Aim 1). We expect integrated progenitors to effectively deliver growth factors to target muscles (including their neuromuscular junctions), thereby preserving motor neuron/muscle attachments, motor neuron survival and limb function. Since the most common cause of death in ALS is respiratory failure, we will further test the hypothesis that diaphragm hSMPC-based growth factor delivery prolongs motor neuron survival, thereby preserving respiratory motor function in ALS rats (Aim 2). Finally, we will create new hSMPC lines from iPSCs derived from familial ALS patient donors. By analyzing their cellular characteristics and co-culturing these cells with motor neurons, we will extend the utility of hSMPCs by simulating ALS in vitro, furthering our understanding of the roles played by muscle derived trophic factors (Aim 3). These aims will provide highly novel insights concerning the potential of ex vivo cell and growth factor-based treatments, and will establish a new disease model to advance our understanding of the relative contributions from muscles and neuromuscular connections in this fatal neurodegenerative disease. iPSC- derived hSMPCs can be used to develop patient-specific, cell-based ALS treatments, and provide novel in vitro models of human disease. The results of this project are expected to accelerate progress towards pre-clinical studies in ALS patients. Given the devastating outcome in ALS, the lack of effective treatments, and the burden on society, it is imperative that the questions posed here be answered in a timely manner.

 描述（由申请人提供）：肌萎缩性侧索硬化症（ALS）是一种进行性疾病，可引起运动神经元变性、肌肉萎缩，最终因呼吸衰竭而死亡。我们在这个项目中的主要目标是确定新建立的人骨骼肌祖细胞/干细胞（hSMPCs）来源于诱导多能干细胞（iPSCs）是否可以用于离体细胞治疗（基于干细胞的生长因子递送），并作为体外模型来研究ALS。指导该提议的基本假设是iPSC衍生的hSMPC有效地分化成新的骨骼肌细胞并有助于肌肉再生。这种能力赋予iPSC衍生的hSMPC递送离体生长因子和体外模拟ALS方面的能力。我们的假设是基于我们发表的工作和新的初步数据，证明了从iPSC生产hSMPC的可行性。我们将制备基因修饰的hSMPC，以提供已知在ALS啮齿动物模型中具有神经保护作用的关键生长因子，包括胶质细胞源性神经营养因子（GDNF）和血管内皮生长因子（VEGF）。在建立细胞后，我们将它们移植到肢体肌肉中，以在ALS大鼠中提供生长因子（目的1）。我们期望整合的祖细胞能有效地将生长因子递送到靶肌肉（包括它们的神经肌肉接头），从而保护运动神经元/肌肉附着、运动神经元存活和肢体功能。由于ALS中最常见的死亡原因是呼吸衰竭，我们将进一步检验基于膈肌hSMPC的生长因子递送促进运动神经元存活，从而保护ALS大鼠的呼吸运动功能的假设（目的2）。最后，我们将从家族性ALS患者供体衍生的iPSC中创建新的hSMPC系。通过分析它们的细胞特性并将这些细胞与运动神经元共培养，我们将通过体外模拟ALS来扩展hSMPCs的效用，进一步了解肌源性营养因子所起的作用（目的3）。这些目标将提供关于离体细胞和生长因子为基础的治疗潜力的高度新颖的见解，并将建立一个新的疾病模型，以促进我们对肌肉和神经肌肉连接在这种致命的神经退行性疾病中的相对贡献的理解。iPSC衍生的hSMPC可用于开发患者特异性的、基于细胞的ALS治疗，并提供新的人类疾病体外模型。该项目的结果有望加速ALS患者临床前研究的进展。鉴于ALS的破坏性后果，缺乏有效的治疗方法以及对社会的负担，必须及时回答这里提出的问题。