Oligodendrocyte Lineage Gene Function in the CNS

少突胶质细胞谱系基因在中枢神经系统中的功能

基本信息

批准号：
8204933
负责人：
DAVID H ROWITCH
金额：
$ 43.24万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-08-01 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8204933
关键词：
Address Affinity Antibodies Avian Sarcoma Avian Sarcoma Viruses Biochemical Biological Cell Cycle Cell Line Cell Nucleus Cells Chromatin Complex DNA Sequence Demyelinating Diseases Developmental Biology Embryo Gene Targeting Genetic Genetic Transcription Growth Infection Laboratories Libraries Light Malignant Glioma Malignant neoplasm of brain Maps Methods Modeling Molecular Motor Neurons Mus Mutant Strains Mice Myelin Neural tube Olig2 protein Oligodendroglia Outcome Patients Pattern Pattern Formation Phospho-Specific Antibodies Phosphorylation Precipitation Preclinical Drug Evaluation Process Protein Kinase Proteins Protocols documentation Reagent Research Role Serine Shivering Spinal Cord Spinal cord injury Stem cells System Testing Transgenic Mice Work base daughter cell dimer expression vector gene function in vivo kinase inhibitor mimetics motor neuron development mouse model mutant myelination nerve stem cell novel oligodendrocyte lineage progenitor protein protein interaction public health relevance scaffold self-renewal single molecule small hairpin RNA small molecule tool transcription factor vector

项目摘要

DESCRIPTION (provided by applicant): The broad objective of the research proposed here is to define the molecular mechanisms that regulate oppositional functions of the bHLH transcription factor Olig2 on proliferation and differentiation of neural progenitor cells. In preliminary studies, we have identified a cluster of three serine residues ("Triple-S motif") in the amino terminus of Olig2 that are phosphorylated in cycling neural progenitors, but not in differentiated progeny. Mutational analysis indicates that phosphorylation of the Olig2 Triple-S motif is required for self-renewal of neural stem cells but is not required for Olig2-dependent specification of immature oligodendrocytes. The work proposed here builds upon these preliminary observations. We have five Specific Aims: Aim One is to identify the protein kinase(s) that phosphorylate Olig2 using small molecule kinase inhibitors and a kinome-wide ShRNAi library. Aim Two is to define Olig2 protein-protein interactions that are regulated by phosphorylation using "TAP- Tagged" wild type, phospho-null and phosphomimetic Olig2 proteins. Aim Three is to define the transcriptional functions of Olig2 phosphorylation to dictate the decision of neural progenitors to self renew or exit the cell cycle and differentiate. We will use ChIP/Seq protocols to identify direct genetic targets of phosphorylated Olig2. Aim Four is to determine a possible requirement for Olig2 Triple-S phosphorylation in embryonic patterning of spinal cord and in motor neuron development using a novel, bifunctional Olig2-tva-cre transgenic mouse neural tube explant system. Aim Five is to determine a possible requirement for Olig2 phosphorylation during oligodendrocyte maturation and CNS tumorgenesis in vivo. Orthotopic grafting studies will determine if Olig2 phosphorylation is essential for oligodendrocyte myelination, or alternatively, tumorgenesis in a mouse model of malignant glioma. PUBLIC HEALTH RELEVANCE: The proposed work may shed light on molecular mechanisms that regulate neural progenitors in malignant gliomas, spinal cord injury and demyelinating diseases.

描述（由申请人提供）：此处提出的研究的广泛目标是定义调节BHLH转录因子Olig2对神经祖细胞增殖和分化的分子机制。在初步研究中，我们已经确定了Olig2氨基末端的三个丝氨酸残基（“ Triple-S基序”）的簇，它们在循环神经祖细胞中被磷酸化，但在分化的后代中没有磷酸化。突变分析表明，olig2 triple-S基序的磷酸化是对神经干细胞的自我更新所必需的，但对于未成熟的少突胶质细胞的olig2依赖性规范并不是必需的。这里提出的作品建立在这些初步观察的基础上。我们有五个特定的目的：目标是鉴定使用小分子激酶抑制剂和整个Kinome Wide shrnai库磷酸化寡磷酸酶的蛋白激酶。目标二是定义使用“ Taps-taked”野生型，磷酸无效和磷酸化的寡聚2蛋白来调节磷酸化的寡蛋白 - 蛋白质相互作用。目的三是定义寡磷酸化的转录功能，以决定神经祖细胞自我更新或退出细胞周期并分化的决定。我们将使用CHIP/SEQ方案来识别磷酸化的Olig2的直接遗传靶标。目的四是使用一种新型的双功能olig2-TVA-TVA-TVA-CRE转基因小鼠神经管epplant系统来确定脊髓和运动神经元发育中寡-2三重S磷酸化的可能要求。目的五是确定在体内少突胶质细胞成熟和中枢神经系统肿瘤发生过程中寡磷酸化的可能要求。原位嫁接研究将确定寡聚2磷酸化是少突胶质细胞髓鞘的必不可少的，还是在恶性神经胶质瘤小鼠模型中肿瘤发生。公共卫生相关性：拟议的工作可能阐明了调节恶性神经胶质瘤，脊髓损伤和脱髓鞘疾病的神经祖细胞的分子机制。