A human neuron/oligodendrocyte co-culture model to study myelination

研究髓鞘形成的人类神经元/少突胶质细胞共培养模型

基本信息

批准号：
8423696
负责人：
BRIDGET SHAFIT-ZAGARDO
金额：
$ 20.14万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8423696
关键词：
Adhesions Agonist Animal Model Area Axon Biological Models Caliber Cannabinoids Cell Maturation Cells Chronic Disease Coculture Techniques Confocal Microscopy Data Demyelinations Disease Electron Microscopy Electrons Goals Growth Growth Factor Receptors Human Human Activities Image In Vitro Incubated Length Maintenance Membrane Methods Microscopic Microscopy Modeling Multiple Sclerosis Myelin Myelin Proteins Myelin Sheath Nature Neurons Oligodendroglia Pathology Patients Process Proteins RXR Recombinants Rodent Role Rotation Sagittaria Series Signal Transduction Signaling Molecule Site Spinal Ganglia System Testing Therapeutic Time alitretinoin cannabinoid receptor in vitro Model insight myelination neurofilament novel oligodendrocyte precursor parallel processing precursor cell protein expression receptor receptor expression remyelination

项目摘要

DESCRIPTION (provided by applicant): A pivotal issue for patients with multiple sclerosis (MS) is the long-term survival and integrity of neurons and oligodendrocytes, and maintenance of the myelin sheath. Over time, the progressive nature of MS results in a chronic disease course with permanent axonal damage, neuronal and oligodendrocyte loss, demyelination, and diminished re-myelination. Studies in animal models have proved useful in exploring disease mechanisms and in testing therapeutic paradigms, however model systems using cells of human origin are clearly essential for the identification of proteins required to maintain and enhance myelination in humans. In this proposal, our goal is to use our human co-culture system of dorsal root ganglia and oligodendrocyte precursor cells (OPC) to determine the extent to which factors proposed to enhance myelination in rodents function similarly in the human system. We will expand upon our preliminary data that show that growth-arrest specific protein 6 (gas6) enhances OPC maturation and axon ensheathment, and test whether in combination with gas6, the activation of cannabinoid receptors, or the retinoid X receptor (RXR) can promote further the maturation and myelinating activity of human OPC in vitro. In Specific Aim 1, we will continue to examine the ability of gas6 to enhance axonal ensheathment, and examine by immunofluorescent microscopy and electron microscopy the extent to which myelin proteins are expressed in the developing myelin sheath, whether compact myelin is achieved over time, and the correlation between axonal diameter and myelin sheath formation. In Specific Aim 2A, we will examine the contribution of agonists and antagonists of cannabinoid (CB) receptors plus and minus gas6 to determine whether CB2 agonists enhance myelination and aid in myelin compaction. In Specific Aim 2B, we will determine whether 9-cis-retinoic acid signaling through the RXR? receptor enhances myelination. These studies will help to define factors required for human myelination. Analysis of growth factors, receptor agonists and signaling molecules will determine the potential role of these factors in enhancing human myelination with a focus on the factors required for compact myelin formation in a human model system.

描述（由申请人提供）：对于多发性硬化症（MS）患者来说，一个关键问题是神经元和少突胶质细胞的长期存活和完整性，以及髓鞘的维护。随着时间的推移，多发性硬化症的进展性会导致慢性病程，伴有永久性轴突损伤、神经元和少突胶质细胞损失、脱髓鞘和髓鞘再生减少。动物模型研究已被证明有助于探索疾病机制和测试治疗范例，然而，使用人类来源细胞的模型系统对于鉴定维持和增强人类髓鞘形成所需的蛋白质显然至关重要。在这项提案中，我们的目标是利用我们的背根神经节和少突胶质细胞前体细胞（OPC）的人类共培养系统来确定增强啮齿类动物髓鞘形成的因子在人类系统中的作用程度。我们将扩展我们的初步数据，这些数据表明生长停滞特异性蛋白 6 (gas6) 增强 OPC 成熟和轴突鞘，并测试与 Gas6 组合，大麻素受体或视黄醇 X 受体 (RXR) 的激活是否可以促进进一步促进人 OPC 的体外成熟和髓鞘形成活性。在具体目标1中，我们将继续研究gas6增强轴突鞘的能力，并通过免疫荧光显微镜和电子显微镜检查髓鞘蛋白在发育中的髓鞘中表达的程度，是否随着时间的推移获得致密的髓鞘，以及轴突直径与髓鞘形成之间的相关性。在特定目标 2A 中，我们将检查大麻素 (CB) 受体激动剂和拮抗剂加和减 Gas6 的作用，以确定 CB2 激动剂是否增强髓鞘形成并帮助髓磷脂压缩。在特定目标 2B 中，我们将确定 9-顺式视黄酸信号传导是否通过 RXR？受体增强髓鞘形成。这些研究将有助于确定人类髓鞘形成所需的因素。对生长因子、受体激动剂和信号分子的分析将确定这些因子在增强人类髓鞘形成中的潜在作用，重点关注人类模型系统中致密髓鞘形成所需的因子。