MOLECULAR CHARACTERIZATION OF NON-MYELINATING SCHWANN CELLS

非髓鞘化雪旺细胞的分子特征

基本信息

批准号：
8804969
负责人：
JEFFREY D MILBRANDT
金额：
$ 25.83万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8804969
关键词：
AIDS/HIV problem Affect Affinity Chromatography Autonomic Dysfunction Axon Biology Burn injury C Fiber Cells Communities Complex Databases Defect Development Diabetes Mellitus Digestion Disease Engineering Exposure to Fiber Functional disorder Gene Deletion Gene Expression Gene Expression Profile Genes Genetic Glean HIV Health Healthcare Inherited Lead Link Malignant Neoplasms Messenger RNA Metabolic Metabolic Diseases Metabolic Pathway Metabolism Methods Mitochondria Modeling Molecular Molecular Profiling Mus Mutation Neuroglia Neuropathy Numbness Orthostatic Hypotension Pain Pathology Patients Peripheral Nerves Peripheral Nervous System Peripheral Nervous System Diseases Play Polyribosomes Proteins Quality of life Research Ribosomal Proteins Ribosomes Role Schwann Cells Sensory Sexual Dysfunction Societies Structure Surveys Symptoms Tamoxifen Therapeutic Agents Transgenic Mice Translating Urination Work care burden cell type chemotherapy diabetic effective therapy functional disability insight mitochondrial dysfunction novel marker recombinase research study tool transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Acquired peripheral neuropathy commonly results from diabetes, chemotherapy and HIV/AIDS, and therefore imposes a considerable, and increasing, health care burden on society. Effective treatment of neuropathy requires an understanding of the interactions between glia and axons in the peripheral nerve. Myelinating Schwann cells have been studied extensively, but less is known about non-myelinating Schwann cells. Because they are associated with unmyelinated sensory fibers that transmit pain, they are likely to participate in the pathology associated with diabetic and other small-fibe neuropathies. Furthermore, Schwann cell-specific mutations in genes that are important for cellular metabolism strongly affect unmyelinated axon stability, revealing an important but little explored link between these cells and peripheral nerve disease. Therefore an understanding of the biology of non-myelinating Schwann cells is an important approach toward developing new treatments for peripheral nerve diseases. We have developed several independent approaches for obtaining gene expression profiles of non-myelinating Schwann cells in order to identify new markers for these cells, and to identify mechanisms whereby their functional impairment in disease, particularly metabolic disease, affects peripheral nerve function. In addition, we have generated transgenic mice that express two proteins, an eGFP-tagged ribosomal protein, and tamoxifen-inducible Cre recombinase, in non-myelinating Schwann cells. The tagged ribosomal protein will permit the isolation and profiling of polyribosome-associated mRNAs in non-myelinating Schwann cells by translating ribosome affinity purification, or TRAP, a powerful method for determining gene expression profiles of specific cell types. The Cre recombinase will permit us to specifically manipulate gene expression in non-myelinating Schwann cells, or ablate them entirely. Such experiments are necessary to dissect the complex interactions between non-myelinating Schwann cells and their associated axons. These new transgenic mouse lines will permit us to establish gene expression signatures of non-myelinating Schwann cells after genetic or environmental perturbation and in models of inherited or acquired PNS disease. In particular we will use these methods to investigate the molecular aspects of how metabolic deficits lead to unmyelinated axon loss and neuropathy. Through the identification and manipulation of genes altered in non-myelinating Schwann cells in health and disease, we hope to gain new insights into the underlying pathology and open up new avenues for treatment of peripheral neuropathies.

描述（由申请人提供）：获得的周围神经病通常是由糖尿病，化学疗法和艾滋病毒/艾滋病引起的，因此给社会带来了相当大的，增加的医疗保健负担。神经病的有效治疗需要了解周围神经中神经胶质和轴突之间的相互作用。已经对髓鞘灌注的细胞进行了广泛的研究，但对非层次的雪旺氏细胞知之甚少。因为它们与传递疼痛的不髓鞘的感觉纤维相关，所以它们很可能参与与糖尿病和其他小型纤维神经病相关的病理。此外，对于细胞代谢很重要的基因中的Schwann细胞特异性突变强烈影响不髓鞘的轴突稳定性，揭示了这些细胞与周围神经疾病之间的重要但很少探索的联系。因此，了解非层状雪旺细胞的生物学是开发新治疗外周神经疾病的重要方法。我们开发了几种独立的方法，用于获得非层状雪旺细胞的基因表达谱，以鉴定这些细胞的新标记，并确定其在疾病中的功能障碍，尤其是代谢性疾病的机制，会影响周围神经功能。此外，我们还产生了表达两种蛋白质的转基因小鼠，一种蛋白质，一种EGFP标记的核糖体蛋白和他莫昔芬可诱导的CRE重物组织酶，在非层状schwann细胞中。标记的核糖体蛋白将允许通过翻译核糖体亲和力纯化或TRAP（一种确定特定细胞类型的基因表达谱的有力方法），在非层化schwann细胞中分离和分析多氧化肌体相关的mRNA。 CRE重组酶将使我们能够在非层状雪旺细胞中专门操纵基因表达，或完全消融它们。这样的实验对于剖析非层状雪旺细胞及其相关轴突之间的复杂相互作用是必要的。这些新的转基因小鼠系将使我们能够在遗传或环境扰动以及遗传或获得的PNS疾病模型中建立非层状雪旺细胞的基因表达特征。特别是，我们将使用这些方法来研究代谢缺陷如何导致无髓轴突丧失和神经病的分子方面。通过鉴定和操纵健康和疾病中非层状雪旺细胞中改变的基因，我们希望获得对潜在病理学的新见解，并为治疗周围神经病的新途径开辟了新的途径。