Novel Gene Targets for CNS Axonal Regeneration

中枢神经系统轴突再生的新基因靶点

基本信息

批准号：
8232574
负责人：
Vance P Lemmon
金额：
$ 2.05万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8232574
关键词：
Adult Affect Animals Axon Bioinformatics Biological Assay Brain Candidate Disease Gene Cell Culture Techniques Cell Transplantation Cells Cicatrix Complementary DNA Complex Cultured Cells Cytoplasmic Granules Data Databases Development Disease Ensure Environment Failure Gene Library Gene Targeting Genes Glycoproteins Grant Hippocampus (Brain)Human In Vitro Individual Injury Interruption Lead Length Libraries Measures Messenger RNA Methods Modeling Molecular Myelin Natural regeneration Neuraxis Neurites Neurons Pathway interactions Phenotype Proteins Proteoglycan RNA Interference Recovery Research Research Personnel Role Screening procedure Signal Pathway Silver Small Interfering RNA Spinal Ganglia Spinal cord injury Stroke System Techniques Testing Transfection Transplantation Trauma Traumatic Brain Injury axon growth axon regeneration base cDNA Library central nervous system injury design functional restoration gene function in vitro Assay in vivo in vivo regeneration inhibitor/antagonist knock-down loss of function neurite growth novel postnatal programs protein protein interaction regenerative research study small hairpin RNA transcription factor white matter

项目摘要

DESCRIPTION (provided by applicant): A major impediment to recovery after spinal cord injury (SCI), traumatic brain injury (TBI), or stroke is the failure of central nervous system (CMS) axons to regenerate effectively through white mater over long distances. A variety of factors are believed to contribute to this problem. These include inhibitors in glial scars, inhibitory material associated with myelin or damaged myelin and molecular changes in neurons during development that reduce their potential for axon growth. Over the past several years it has been shown that Dorsal Root Ganglion (DRG) neurons can send axons very long distances in white mater if they are transplanted into the CNS using techniques that minimize damage. In contrast, transplanting CNS neurons the same way does not produce the same result; i.e. they fail to send out long axons through white matter tracks. This implies that DRG neurons and CNS neurons have inherent molecular differences that limit CNS regenerative efficiency. We propose to test a specific hypothesis that DRG neurons express different genes than CNS neurons, which permit DRG neurons to regeneration in the CNS. In specific aim 1 we will identify these molecular differences using serial subtraction of cDNA libraries. We will look for unique genes in DRG neurons that might enhance regeneration and unique genes in hippocampal neurons and corticospinal neurons that might inhibit regeneration This method is extremely effective at identifying rare and perhaps novel cDNAs, ensuring identification of important targets, such as transcription factors. We will also search public microarray databases to search for additional candidates. In specific aim 2 candidate genes will be tested using a well-established in vitro assay where neurons are grown on myelin or proteoglycans and the lengths of their neurites measured. CNS neurons will be transfected with DRG specific genes or use RNAi of CNS specific genes in order to evaluate the target genes roles in axon growth on inhibitory substrates. Specific aim 3 will use neuronal transfection and microtransplantation in vivo. This will permit us to directly test the role of each candidate gene in the most relevant assay, regeneration in the mammalian central nervous system. These experiments will provide entirely new information about the proteins expressed in DRG neurons that allows them to extend long axons in the CNS. The identification of these targets and testing multiple candidates using in vitro methods and subsequently a refined subset using in vivo approaches should lead directly to potential treatments for SCI, TBI and stroke. Lay Summary: These experiments are designed to identify genes involved in controlling regeneration in white matter in the adult brain. The genes will be tested in neurons that cannot normally grow axons using cell culture and transplantation into animals to determine if the genes promote regeneration.

描述（由申请人提供）：脊髓损伤（SCI），创伤性脑损伤（TBI）或中风后恢复的主要障碍是中枢神经系统（CMS）轴突在长距离长距离上通过白色母校有效再生的失败。据信多种因素会导致这个问题。这些包括神经胶质疤痕的抑制剂，与髓磷脂或髓磷脂受损的抑制材料以及发育过程中神经元的分子变化，从而降低了其轴突生长的潜力。在过去的几年中，已经表明，如果使用最小化损伤的技术将背部根神经节（DRG）神经元移植到中枢神经系统中，则可以在白色孕妇中发送轴突很长。相反，以相同方式移植CNS神经元不会产生相同的结果。即，他们无法通过白色物质轨道发送长轴。这意味着DRG神经元和CNS神经元具有固有的分子差异，限制了CNS再生效率。我们建议检验一个特定的假设，即DRG神经元与CNS神经元表达不同的基因，后者允许DRG神经元在CNS中再生。在特定目标1中，我们将使用cDNA文库的串行减法来确定这些分子差异。我们将寻找DRG神经元中的独特基因，这些基因可能会增强可能抑制再生的海马神经元和皮质脊髓神经元中的再生和独特的基因，在确定稀有且可能是新颖的cDNA中，确保鉴定重要靶标，例如转录因子，例如。我们还将搜索公共微阵列数据库以搜索其他候选人。在特定的目标中，将使用良好的体外测定法测试候选基因，其中神经元在髓磷脂或蛋白聚糖上生长，并测量其神经突的长度。 CNS神经元将用DRG特异性基因转染或使用CNS特定基因的RNAi，以评估轴突生长在抑制性底物上的靶基因作用。特定的目标3将在体内使用神经元转染和微移植。这将使我们能够直接测试每个候选基因在哺乳动物中枢神经系统中最相关的测定中的作用。这些实验将提供有关DRG神经元中表达的蛋白质的全新信息，这些信息使它们可以扩展中枢神经系统中的长轴突。使用体外方法对这些靶标的鉴定并测试多个候选物，随后使用体内方法进行精制子集，应直接导致SCI，TBI和中风的潜在治疗方法。摘要摘要：这些实验旨在识别与成人大脑中白质再生有关的基因。这些基因将在无法使用细胞培养并移植到动物的神经元中测试，以确定基因是否促进再生。