Stress resistance in neurons from primate iPS cells

灵长类 iPS 细胞神经元的应激抵抗力

• 批准号: 8720662
• 负责人: PETER J HORNSBY
• 金额: $ 7.48万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8720662
• 关键词: Adverse effects; Aging; Applications Grants; Biomedical Research; Callithrix; Cardiac Myocytes; Cell Death; Cell Line; Cells; Cholinergic Receptors; Coculture Techniques; Comparative Biology; Comparative Study; Complex; Dendrites; Dendritic Spines; Dependence; Derivation procedure; Drug usage; Ethics; Exhibits; Fibroblasts; Generations; Glucose; Health; Human; Hyperglycemia; In Vitro; Intervention; Legal; Longevity; Maintenance; Measures; Mediation; Metabolism; Mitochondria; Molecular; Monitor; Morphology; Motor Neurons; Muscle Cells; Muscle Fibers; Nerve Degeneration; Neuroectoderm; Neuromuscular Junction; Neurons; Neurophysiology - biologic function; Oxidative Stress; Oxygen; Pan Genus; Pathway interactions; Phase; Physiological; Play; Positioning Attribute; Primates; Property; Protocols documentation; Quality Control; Resistance; Role; Skin; Somatic Cell; Stress; Sulfhydryl Compounds; Synapses; System; Testing; Time; Trees; Vertebral column; cell type; density; embryonic stem cell; experience; in vivo; induced pluripotent stem cell; insight; interest; mitogen-activated protein kinase p38; molecular marker; motor neuron development; neural patterning; neuron apoptosis; nonhuman primate; research study; species difference

DESCRIPTION (provided by applicant): Induced pluripotent stem cells (iPS cells) have properties similar to those of embryonic stem cells, but they can be derived from any type of somatic cell, such as a skin fibroblast. iPS cells can provide access to specialized cells, such as neurons, in species that otherwise would be unavailable for biomedical research. Among nonhuman primates, the chimpanzee occupies a unique position for comparative biology. Here we propose to use iPS cells from three primates: the chimpanzee, marmoset, and human. Few studies have yet taken advantage of the unique properties of iPS cells for comparative studies of mammalian aging. Our hypothesis is that differentiated cells (motor neurons) derived from three primate species of very different longevities will exhibit differential resistance to physiological stresses, and that pharmacological interventions in key pathways involved in stress resistance will reveal the extent to which mechanisms of stress resistance differ among neurons of these three primate species. Specific Aim 1: To validate conditions for efficient and robust derivation of motor neurons from chimpanzee, marmoset, and human iPS cells. We will derive motor neurons from these three primate species using a three-phase protocol (induction of neuroectoderm, neural patterning, and motor neuron development/maturation), by adapting protocols that have been used for efficient differentiation in human pluripotent cells. We hypothesize that motor neurons from all species will be capable of forming typical neuromuscular junctions (NMJs) when co-cultured with a skeletal muscle cell line. Specific Aim 2: To assess whether stress resistance in motor neurons of three primate species varies proportionally to the different longevities of these three species. We will use elevated glucose and elevated oxygen as in vitro conditions that mimic the long-term stresses that neurons may experience in vivo. We hypothesize that robustness of neurons under conditions of stress will vary in proportion to the differing life spans of the primate species. We further hypothesize that quality control mechanisms in mitochondria, at the molecular and organellar levels, will be a major determinant of stress resistance. Specific Aim 3. To assess the potential for pharmacological interventions in key pathways to ameliorate the effects of stresses in motor neurons of the three primate species. While stress resistance is complex, there are three key pathways: NF-?B, p38 MAP kinase, and cell thiol metabolism, that have been implicated in the adverse effects of oxidative stress and hyperglycemia on neural function, or in cellular protective mechanisms. Using drugs that have been well established to act on these three pathways, we will assess whether these interventions influence the adverse effects of stresses in cultured motor neurons. These experiments will allow, for the first time, direct comparisons of stress resistance in isolated neurons from different species and will provide new insights into the mechanisms by which species differ in longevity.

描述(申请人提供)：诱导多能干细胞(iPS细胞)具有与胚胎干细胞相似的特性，但它们可以来自任何类型的体细胞，如皮肤成纤维细胞。IPS细胞可以提供对特定细胞的访问，例如 神经元，否则将无法用于生物医学研究的物种。在非人类灵长类动物中，黑猩猩在比较生物学中占有独特的地位。在这里，我们建议使用来自三种灵长类动物的iPS细胞：黑猩猩、绒猴和人类。很少有研究利用iPS细胞的独特特性来比较研究哺乳动物的衰老。我们的假设是，来自三种寿命迥异的灵长类动物的分化细胞(运动神经元)将对生理应激表现出不同的抗性，而对参与应激反应的关键途径的药物干预将揭示这三种灵长类动物神经元的应激抗性机制的不同程度。具体目标1：验证从黑猩猩、绒猴和人类iPS细胞高效而稳健地获得运动神经元的条件。我们将使用三个阶段的方案(神经外胚层的诱导、神经模式的形成和运动神经元的发育/成熟)从这三个灵长类物种中提取运动神经元，方法是采用已被用于在人类多能细胞中有效分化的方案。我们假设，当所有物种的运动神经元与骨骼肌细胞系共培养时，都能够形成典型的神经肌肉连接(NMJ)。具体目标2：评估三种灵长类动物运动神经元的应激抗性是否与这三种动物不同的寿命成正比。我们将使用升高的葡萄糖和升高的氧气作为体外条件，模拟神经元在体内可能经历的长期压力。我们假设，神经元在应激条件下的健壮性将随着灵长类物种不同的寿命而成比例地变化。我们进一步假设，线粒体在分子和细胞器水平上的质量控制机制将是抗逆性的主要决定因素。具体目的3.评估在关键通路上进行药物干预以改善三种灵长类动物运动神经元应激反应的可能性。虽然抗逆性是复杂的，但有三个关键途径：核因子？？b、p38蛋白激酶和细胞硫醇代谢，与氧化应激和高血糖对神经功能或细胞保护的不利影响有关。 机制。我们将使用已经确定的作用于这三条通路的药物，评估这些干预措施是否会影响培养的运动神经元中应激的不利影响。这些实验将首次对不同物种分离的神经元的抗应激能力进行直接比较，并将为不同物种寿命差异的机制提供新的见解。