Metabolic Control of Proliferation and Differentiation in Oligodendrocytes

少突胶质细胞增殖和分化的代谢控制

• 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
• 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

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 治疗。该奖学金探讨了新陈代谢的作用，更具体地说是葡萄糖衍生的乙酰基的作用 CoA (AcCoA)，调节发育中的髓鞘形成。实验目标建立在以下坚实的前提之上： AcCoA 是一种不稳定的化合物，不能自由地从一个隔室扩散到下一个隔室。首要的 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) 以及内质网特异性 AcCoA 转运蛋白 SLC33A1 的水平。目的 1 使用功能丧失和功能获得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 祖状态。它还假设出生后第二周血糖的短暂下降是 导致核 ACLY 减少、核 AcCoA 减少，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。该假设将使用 Acly 损失和 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。基因组 将使用染色质免疫沉淀来测试所选组蛋白乙酰化标记的广泛分布。目标2 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。这一假设得到了小鼠髓鞘质增加的检测的支持。 Slc33a1 转基因的系统性过度表达。这些子目标将解决 OL 分化和髓磷脂问题 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。这 培训计划包括学习新技能，例如表观遗传学、生物信息学、 慢病毒转导、光遗传学和最新的质谱成像技术。此外， 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 会议将允许建立联系。作为切实的里程碑，这项工作预计将产生两个高质量的成果： 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。该奖学金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 科学家和管理员在我们对大脑发育和疾病的理解方面取得了进展。