MOLECULAR CHARACTERIZATION OF NON-MYELINATING SCHWANN CELLS

非髓鞘化雪旺细胞的分子表征

基本信息

批准号：
8679902
负责人：
JEFFREY D MILBRANDT
金额：
$ 29.62万
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/8679902
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 public health relevance recombinase research study tool

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）：获得性周围神经病通常由糖尿病、化疗和艾滋病毒/艾滋病引起，因此给社会带来了相当大且不断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胶质细胞和轴突之间的相互作用。有髓鞘雪旺细胞已被广泛研究，但对非髓鞘雪旺细胞知之甚少。因为它们与传递疼痛的无髓鞘感觉纤维有关，所以它们很可能参与与糖尿病和其他小纤维神经病相关的病理学。此外，对细胞代谢很重要的雪旺细胞特异性基因突变强烈影响无髓鞘轴突的稳定性，揭示了这些细胞与周围神经疾病之间的重要但很少被探索的联系。因此，了解非髓鞘雪旺细胞的生物学是开发周围神经疾病新疗法的重要方法。我们开发了几种独立的方法来获取非髓鞘雪旺细胞的基因表达谱，以便识别这些细胞的新标记，并确定它们在疾病（特别是代谢疾病）中的功能损伤影响周围神经功能的机制。此外，我们还培育出了在非髓鞘雪旺细胞中表达两种蛋白质的转基因小鼠，一种是 eGFP 标记的核糖体蛋白，另一种是他莫昔芬诱导的 Cre 重组酶。标记的核糖体蛋白将允许通过翻译核糖体亲和纯化 (TRAP) 来分离和分析非髓鞘化雪旺细胞中的多核糖体相关 mRNA，TRAP 是一种确定特定细胞类型基因表达谱的强大方法。 Cre 重组酶将使我们能够特异性地操纵非髓鞘雪旺细胞中的基因表达，或完全消除它们。这些实验对于剖析非髓鞘雪旺细胞与其相关轴突之间复杂的相互作用是必要的。这些新的转基因小鼠品系将使我们能够在遗传或环境扰动后以及遗传性或获得性三七总皂苷疾病模型中建立非髓鞘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鞘轴突丢失和神经病变的分子方面。通过鉴定和操作健康和疾病中非髓鞘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，并为周围神经病的治疗开辟新途径。