mTOR complexes in oligodendrocyte differentiation

mTOR 复合物在少突胶质细胞分化中的作用

基本信息

批准号：
8704546
负责人：
Stacey Elizabeth Wahl
金额：
$ 4.22万
依托单位：
RBHS-NEW JERSEY MEDICAL SCHOOL
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：
2012-07-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8704546
关键词：
Actins Action Potentials Adaptor Signaling Protein Adult Affect Axon Cell Differentiation process Cell Proliferation Cell physiology Cellular Morphology Complex Corpus Callosum Cytoskeletal Proteins Cytoskeleton DNA Binding Data Demyelinations Development Electron Microscopy Family member G-Protein-Coupled Receptors GPR17 gene Gene Expression Genetic Transcription Goals Immunohistochemistry In Vitro Injury Laboratories Lesion Lysophosphatidylcholines Mediating Messenger RNA Multiple Sclerosis Mus Myelin Myelin Basic Proteins Myelin Proteins Myelin Sheath Neuraxis Oligodendroglia Pathology Pathway interactions Platelet Factor 4 Population Post-Translational Protein Processing Process Production Proteins Proteomics Proto-Oncogene Proteins c-fyn Raptors Rattus Regulation Reporter Role Signal Transduction Sirolimus Small Interfering RNA Spinal Cord Staging Stem cells System Testing Therapeutic Time Transcriptional Regulation Transgenes Transgenic Model YY1 Transcription Factor Yin-Yang base cell growth cell type in vivo in vivo Model inhibitor/antagonist mTOR protein myelination neonatal hypoxic-ischemic brain injury neuropathology oligodendrocyte-myelin glycoprotein postnatal progenitor protein expression remyelination response therapeutic target transcription factor

项目摘要

DESCRIPTION (provided by applicant): Oligodendrocytes envelop axons with a myelin sheath in the central nervous system to facilitate action potential conduction. The establishment of axon myelination during development involves oligodendrocyte proliferation, differentiation, and production of myelin. The intracellular signaling mechanisms governing this process are only beginning to be elucidated. Previous data in our laboratory demonstrated mammalian target of rapamycin (mTOR) as essential for oligodendrocyte differentiation. mTOR, a downstream target of PI3K/Akt, regulates cell growth and proliferation in a number of cell types through complexes with adaptor proteins raptor (mTORC1) or rictor (mTORC2). mTORC1 and mTORC2 are present in oligodendrocyte progenitors and previous data indicate different functions for each complex during differentiation. My hypothesis is that the two mTOR complexes have essential and distinct functions during oligodendrocyte differentiation and that mTOR signaling is essential for remyelination as well as developmental myelination. Through the use of siRNA knockdown of complex specific proteins in primary oligodendrocyte cultures the effects of knockdowns of mTORC1 and mTORC2 will be characterized in vitro. Bigenic mice carrying floxed-mTOR and either a CNP-Cre transgene or an inducible PLP-Cre transgene will be created for oligodendrocyte specific knockdown of mTOR in vivo. These systems will be used to investigate the effects of specific deletion of mTOR on differentiation and myelination. Completion of these studies will elucidate how mTOR functions in oligodendrocyte differentiation, myelination and remyelination in vivo. The first aim will investigate the role of mTORC2 in the regulation of transcription factors integral to the differentiation process. Aim 2 will assess mTORC2 regulation of the cytoskeleton and process outgrowth. Aim 3 will utilize in vivo models to determine the effects of mTOR knockdown on differentiation and myelination during development and following a demyelinating injury. Oligodendrocyte differentiation is disrupted in neonatal hypoxia-ischemia and in Multiple Sclerosis, illustrating a need for a detailed understanding of the mechanistic processes involved. These studies will further the understanding of oligodendrocyte differentiation and provide targets to pursue for therapeutic treatments of neuropathologies that impact oligodendrocytes and myelin formation.

描述（由申请人提供）：中枢神经系统中带有髓鞘鞘的少突胶质细胞包膜，以促进动作潜在传导。发育过程中轴突髓鞘形成的建立涉及髓磷脂的少突增殖，分化和产生。管理此过程的细胞内信号传导机制才刚刚开始阐明。我们实验室中的先前数据表明，雷帕霉素（MTOR）的哺乳动物靶标对于少突胶质细胞分化至关重要。 MTOR是PI3K/AKT的下游靶标，可以通过与衔接蛋白Raptor（MTORC1）或Rictor（MTORC2）的复合物（MTORC1）调节许多细胞类型的细胞生长和增殖。 MTORC1和MTORC2存在于少突胶质细胞祖细胞中，并且先前的数据表明在分化过程中，每个复合物的功能都不同。我的假设是，这两个MTOR复合物在少突胶质细胞分化过程中具有必不可少的和不同的功能，并且MTOR信号传导对于延迟和发育性髓鞘化至关重要。通过在原代寡胶质细胞培养物中使用复杂特异性蛋白的siRNA敲低，MTORC1和MTORC2的敲低的影响将在体外表征。携带Floxed-MTOR和CNP-CRE转基因或可诱导的PLP-CRE转基因的胆汁小鼠将用于体内MTOR的少突胶质细胞特异性敲低。这些系统将用于研究MTOR特定缺失对分化和髓鞘化的影响。这些研究的完成将阐明MTOR在体内的少突胶质分化，髓鞘化和再髓的功能。第一个目的将研究MTORC2在调节分化过程中不可或缺的转录因子中的作用。 AIM 2将评估细胞骨架和过程产物的MTORC2调节。 AIM 3将利用体内模型来确定MTOR敲低对发育过程中分化和髓鞘形成的影响，并在脱髓鞘损伤之后。在新生儿缺氧 - 缺血性和多发性硬化症中，少突胶质细胞分化被破坏，这说明了对所涉及的机械过程的详细了解。这些研究将进一步了解少突胶质细胞分化，并为影响少突胶质细胞和髓磷脂形成的神经病理的治疗治疗提供靶标。