Metabolic Control of Proliferation and Differentiation in Oligodendrocytes

少突胶质细胞增殖和分化的代谢控制

基本信息

批准号：
10315354
负责人：
Sami Sauma
金额：
$ 3.17万
依托单位：
ADVANCED SCIENCE RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10315354
关键词：
ATP Citrate (pro-S)-Lyase Ablation Acetyl Coenzyme A Address Appointment Area Bioinformatics Biology Brain Cell Cycle Cell Differentiation process Cell Nucleus ChIP-seq Cholesterol Communication Cytosol Data Data Analyses Detection Development Diet Differentiated Gene Diffuse Disease Electron Microscopy Elements Endoplasmic Reticulum Epigenetic Process Experimental Designs Fatty Acids Fellowship Fostering Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Transcription Genome Glucose Goals Grant Histone Acetylation Histone Deacetylation Histones Image Imaging technology Impairment In Vitro Institution International Lead Learning Light Lipids Maintenance Manuscripts Mass Spectrum Analysis Membrane Mental disorders Mentors Metabolic Metabolic Control Metabolism Methodology Mission Molecular Mus Myelin Myelin Sheath National Institute of Neurological Disorders and Stroke Nuclear Oligodendroglia Oral Outcome Pharmacology Phenotype Postdoctoral Fellow Process Proliferating Public Health Publications Publishing Regulation Reporting Research Rest Role Science Scientist Solid Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Students System Testing Training Transcript Transgenes Transgenic Organisms Undifferentiated United States National Institutes of Health Work base brain cell career cdc Genes chromatin immunoprecipitation epigenetic regulation gain of function genome-wide in vivo inhibitor/antagonist insight loss of function metabolic abnormality assessment myelination nervous system disorder novel therapeutics oligodendrocyte progenitor optogenetics overexpression postnatal progenitor repaired skills stem cells symposium synthetic enzyme transcriptome sequencing undergraduate student white matter

项目摘要

Project Summary Myelin is critical for proper brain function and its dysfunction, damage or inappropriate formation has been reported in a wide range of neurological and psychiatric disorders, thereby urging the discovery of new potential treatments. This fellowship addresses the role of metabolism, and more specifically of glucose-derived acetyl- CoA (AcCoA), in regulating developmental myelination. The experimental aims rest on the solid premise that AcCoA is an unstable compound which cannot freely diffuse from one compartment to the next. The overarching hypothesis is that AcCoA function is dependent on the subcellular localization of its synthetic enzyme ATP citrate lyase (ACLY) and on the levels of the specific AcCoA transporter to the endoplasmic reticulum, SLC33A1. Aim 1 uses loss- and gain-of-function approaches to test the hypothesis that high glucose levels during the first postnatal week favor nuclear localization of ACLY and in turn promote synthesis of AcCoA and its incorporation into histones, thereby resulting in the expression of genes that favor proliferation and the maintenance of the progenitor state. It also posits that the transient decline of glucose during the second postnatal week is responsible for decreased nuclear ACLY, decreased nuclear AcCoA thereby favoring histone deacetylation and the transition of OPC from proliferating to differentiating cells. The hypothesis will be tested using Acly loss- and gain-of-function approaches in vitro in cultured OPC as well as lineage specific ablation in mice. The genome wide distribution of select histone acetylation marks will be tested using chromatin immunoprecipitation. Aim 2 uses loss- and gain-of-function approaches to test the hypothesis that increased cytosolic AcCoA synthesis in differentiating OL, followed by its transport to the endoplasmic reticulum (via SLC33A1) is crucial for the synthesis of cholesterol and myelin lipids. This hypothesis is supported by the detection of increased myelin in mice with systemic overexpression of the Slc33a1 transgene. The subaims will address OL differentiation and myelin development by matrix assisted laser desorption/ionization (MALDI) imaging, and electron microscopy. The training plan incorporates learning of new skills, such as advanced methodology in epigenetics, bioinformatics, lentiviral transduction, optogenetics and the latest in mass spectrometry imaging technologies. In addition, several opportunities will be offered to encourage training in experimental design, data analysis, as well as improvements in written and oral scientific communication and opportunities to mentor undergraduate students. Professional development opportunities will be available and participation in local, national, and international conferences will allow networking. As tangible milestones, the work is expected to result in two high quality, first author manuscripts as a doctoral trainee, and grant the opportunity to obtain a competitive post-doctoral appointment, leading to a career in academic science. This fellowship aligns the applicant’s long-term goal of studying metabolic regulation of brain cells, with the public health mission of NIH and NINDS to foster academic scientists and steward advances in our understanding of brain development and disease.

项目概要髓磷脂对于大脑的正常功能至关重要，其功能障碍、损伤或不适当的形成已被证实已报道了广泛的神经和精神疾病，从而敦促发现新的潜力该研究金研究了代谢的作用，更具体地说是葡萄糖衍生的乙酰基的作用。 CoA (AcCoA)，在调节发育髓鞘形成方面的实验目标基于以下坚实的前提： AcCoA 是一种不稳定的化合物，不能自由地从一个隔室扩散到另一个隔室。假设 AcCoA 功能依赖于其合成酶 ATP 柠檬酸的亚细胞定位裂解酶 (ACLY) 和内质网特异性 AcCoA 转运蛋白 SLC33A1 的水平。 1 使用功能丧失和获得的方法来检验以下假设：第一阶段的高血糖水平产后一周有利于 ACLY 的核定位，进而促进 AcCoA 的合成及其掺入成组蛋白，从而导致有利于增殖和维持的基因的表达它还假设出生后第二周血糖的短暂下降是负责核减少 ACLY 减少，核 AcCoA 从而有利于组蛋白脱乙酰化和 OPC 从增殖细胞向分化细胞的转变将使用 Acly 丢失和分化进行检验。体外培养的 OPC 中的功能获得方法以及小鼠中的谱系特异性消融。将使用染色质免疫沉淀法测试广泛分布的选定组蛋白乙酰化标记。使用功能丧失和获得的方法来检验以下假设：细胞质 AcCoA 合成增加分化 OL，然后将其转运至内质网（通过 SLC33A1）对于合成至关重要胆固醇和髓磷脂脂质的增加得到了小鼠髓磷脂增加的检测的支持。 Slc33a1 转基因的系统性过度表达。子目标将解决 OL 分化和髓鞘质问题。通过基质辅助激光解吸/电离（MALDI）成像和电子显微镜进行开发。培训计划包括学习新技能，例如表观遗传学、生物信息学、此外，慢病毒转导、光遗传学和最新的质谱成像技术。将提供一些机会来鼓励实验设计、数据分析以及书面和口头科学交流的改善以及指导本科生的机会。将提供专业发展机会并参与地方、国家和国际活动作为切实的里程碑，这项工作预计将产生两个高质量的结果：第一。作为博士生实习生撰写稿件，并获得获得有竞争力的博士后的机会任命，导致学术科学的职业生涯与申请人的长期目标相一致。研究脑细胞的代谢调节，以 NIH 和 NINDS 的公共卫生使命促进学术发展科学家和管理者对大脑发育和疾病的理解。