Role of Pacs2 in central nervous system myelination

Pacs2在中枢神经系统髓鞘形成中的作用

• 批准号: 10284160
• 负责人: YUNGKI PARK
• 金额: $ 43.86万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 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 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10284160
• 关键词: 4-Hydroxy-Tamoxifen; Acute; Address; Alleles; Axon; Behavior; Biological; Brain; CRISPR interference; Categories; Cell Differentiation process; Cell Lineage; Cells; Cerebellum; Cervical; Control Groups; Corpus Callosum; Corpus striatum structure; Development; Endoplasmic Reticulum; Ethanol; Event; Exhibits; Functional disorder; Gene Expression; Genes; Growth; Hand Strength; Homeostasis; Hour; Human; Impairment; In Vitro; International; Intracellular Transport; Knock-out; Knockout Mice; Link; LoxP-flanked allele; Mitochondria; Molecular; Motor; Mus; Mutation; Myelin; Myelin Sheath; Neuraxis; Neurodevelopmental Disorder; Neurologic; Oligodendroglia; Ontology; Optic Nerve; Pathogenesis; Phenotype; Platelet-Derived Growth Factor alpha Receptor; Play; Process; Protein Sortings; Proteins; RNA; Reflex action; Role; Sorting - Cell Movement; Spinal Cord; Synapses; Syndrome; Testing; Tissues; Transcript; autism spectrum disorder; conditional knockout; dysmyelination; experimental study; in vivo; insight; knock-down; mouse model; myelination; nervous system disorder; neuropsychiatric disorder; novel; oligodendrocyte lineage; oligodendrocyte precursor; precursor cell; skills; transcription factor; transcriptome sequencing

Project Summary Myelination of the central nervous system (CNS) by oligodendrocytes (OLs) is essential for the development and function of the CNS. Myelin develops in the CNS as OL precursor cells (OPCs) differentiate into OLs, highlighting OL differentiation as a key event for CNS myelination. Myrf is a master transcription factor of OL differentiation and CNS myelination. Conditional knockout (cKO) of Myrf in OL lineage cells leads to complete arrest of OL maturation and lethal dysmyelination. Apparently, the indispensable role of Myrf reflects that of Myrf target genes in the myelination process. To find novel Myrf targets that may play a critical role in OL development, we performed an RNA-seq experiment where we acutely knocked out Myrf in differentiating OLs. As expected, the expression of many genes that are crucial for OL differentiation and CNS myelination went down significantly upon acute Myrf knockout. Unexpectedly, however, a gene ontology analysis revealed that Myrf specifically activates the expression of genes related to cell projection, synapse, and intracellular transport in pre-myelinating OLs. Further, it suggested that molecular machineries involved in axon elongation may propel the growth of OL processes, which contact axons and eventually form myelin sheaths. Studying genes that belong to these categories may provide a novel insight into OL differentiation and CNS myelination. In this regard, we decided to focus on Pacs2 (phosphofurin acidic cluster sorting protein 2), which was significantly down-regulated upon acute Myrf knockout. Pacs2 is a multifunctional protein that plays an important role in diverse processes such as endoplasmic reticulum-mitochondria contact and cargo sorting and transport. PACS2 mutations have been linked to human neurodevelopmental disorders, including a syndromic form of autism. Remarkably, Pacs2 is mainly expressed by OLs in the CNS. Together with the RNA-seq finding that Pacs2 is a putative Myrf target, these observations suggest that Pacs2 may be required for OL development and CNS myelination and that PACS2 dysfunction in OL lineage cells may contribute to the pathogenesis of autism. In support of these hypotheses, our preliminary study found that Pacs2 is required for the in vitro differentiation of mouse OPCs, and Pacs2 expression was significantly downregulated in several autism mouse models where myelin deficiency was observed. Moreover, Pacs2 whole-body knockout mice exhibited neurological phenotypes. Currently, little is known about the role of Pacs2 in in vivo OL development and CNS myelination. This project intends to address it by deleting Pacs2 in OL lineage cells through the Cre- loxP approach.

项目概要 少突胶质细胞 (OL) 对中枢神经系统 (CNS) 的髓鞘化对于发育至关重要 和中枢神经系统的功能。当 OL 前体细胞 (OPC) 分化为 OL 时，髓磷脂在 CNS 中发育， 强调 OL 分化是中枢神经系统髓鞘形成的关键事件。 Myrf 是 OL 的主要转录因子 分化和中枢神经系统髓鞘形成。 OL 谱系细胞中 Myrf 的条件性敲除 (cKO) 导致完全 OL 成熟停滞和致命的髓鞘形成障碍。显然，Myrf 不可或缺的角色反映了 Myrf 髓鞘形成过程中的靶基因。寻找可能在 OL 中发挥关键作用的新 Myrf 靶点 开发过程中，我们进行了一项 RNA-seq 实验，在区分 OL 的过程中，我们敏锐地敲除了 Myrf。 正如预期的那样，许多对 OL 分化和 CNS 髓鞘形成至关重要的基因的表达发生了变化。 急性 Myrf 敲除后显着下降。然而出乎意料的是，基因本体分析显示， Myrf 特异性激活与细胞投射、突触和细胞内相关的基因的表达 髓鞘形成前 OL 中的运输。此外，它表明参与轴突伸长的分子机器 可能会促进 OL 过程的生长，OL 过程接触轴突并最终形成髓鞘。学习中 属于这些类别的基因可能为 OL 分化和 CNS 髓鞘形成提供新的见解。 在这方面，我们决定重点关注Pacs2（磷酸呋喃酸性簇分选蛋白2），它是 急性 Myrf 敲除后显着下调。 Pacs2 是一种多功能蛋白质，具有 在内质网-线粒体接触和货物分类等多种过程中发挥重要作用 运输。 PACS2 突变与人类神经发育障碍有关，包括综合征 自闭症的形式。值得注意的是，Pacs2 主要由 CNS 中的 OL 表达。结合 RNA-seq 发现 Pacs2 是假定的 Myrf 目标，这些观察结果表明 OL 可能需要 Pacs2 OL谱系细胞中的PACS2功能障碍可能导致发育和中枢神经系统髓鞘形成 自闭症的发病机制。为了支持这些假设，我们的初步研究发现 Pacs2 是 小鼠OPCs的体外分化，在一些小鼠中Pacs2表达显着下调 观察到髓磷脂缺乏的自闭症小鼠模型。此外，Pacs2全身敲除小鼠 表现出神经表型。目前，人们对 Pacs2 在体内 OL 发育中的作用知之甚少 和中枢神经系统髓鞘形成。该项目打算通过 Cre- 删除 OL 谱系细胞中的 Pacs2 来解决这个问题。 loxP 方法。