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（由申请人提供）：诱导的多能干细胞（IPS细胞）具有与胚胎干细胞相似的特性，但它们可以源自任何类型的体细胞，例如皮肤成纤维细胞。 IPS细胞可以提供对专业单元的访问，例如 神经元，在生物医学研究中不可用的物种中。在非人类灵长类动物中，黑猩猩占据了比较生物学的独特位置。在这里，我们建议使用来自三个灵长类动物的IPS细胞：黑猩猩，泥泞和人类。很少有研究尚未利用IPS细胞的独特特性来进行哺乳动物衰老的比较研究。我们的假设是，源自三种具有不同寿命的灵长类动物物种的分化细胞（运动神经元）将表现出对生理压力的不同耐药性，并且在压力抗性的关键途径中的药理干预措施将揭示这三种灵长类动物物种之间应力抗性差异的程度。具体目标1：验证从黑猩猩，果泥和人IPS细胞中有效且可靠的运动神经元的有效衍生的条件。我们将使用三相方案（神经外胚层的诱导，神经模式和运动神经元的发育/成熟）从这三个灵长类动物物种中得出运动神经元，该方案通过适应已用于有效分化人类多能细胞的方案。我们假设所有物种的运动神经元将能够与骨骼肌细胞系共培养时形成典型的神经肌肉连接（NMJ）。具体目的2：评估三种灵长类动物物种的运动神经元中的压力抗性是否与这三种物种的不同寿命成正比。我们将使用升高的葡萄糖和升高的氧气作为体外条件，这些条件模仿了神经元可能在体内经历的长期应力。我们假设在压力条件下，神经元的鲁棒性与灵长类动物物种的不同寿命成正比。我们进一步假设线粒体的质量控制机制在分子和细胞器水平上将是应激抗应力的主要决定因素。具体目的3。评估关键途径中药理干预的潜力，以改善三种灵长类动物物种运动神经元的应激作用。虽然应力抗性是复杂的，但有三个关键途径：NF-？B，P38 MAP激酶和细胞硫醇代谢，与氧化应激和高血糖对神经功能的不良影响有关，或者与细胞保护性有关 机制。使用已建立的药物对这三种途径作用，我们将评估这些干预措施是否影响培养的运动神经元中应力的不利影响。这些实验将首次允许在不同物种的孤立神经元中直接比较应力抗性，并将为物种在寿命上不同的机制提供新的见解。