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.

项目摘要 少突胶质细胞(OLs)对中枢神经系统(CNS)的髓鞘形成是发育所必需的 和中枢神经系统的功能。随着OL前体细胞(OPC)分化为OL，髓鞘在中枢神经系统发育， 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系细胞中Myrf的条件性基因敲除(CKO)导致完成 OL成熟受阻和致死性髓鞘萎缩。显然，米尔夫不可或缺的作用反映了 Myrf在髓鞘形成过程中的靶基因。寻找在OL中可能起关键作用的新的Myrf靶点 在开发过程中，我们进行了RNA-SEQ实验，在该实验中，我们在区分OL的过程中敏锐地敲除了Myrf。 正如预期的那样，许多对OL分化和中枢神经系统髓鞘形成至关重要的基因的表达 在急性Myrf基因敲除后显著下降。然而，出乎意料的是，一项基因本体论分析揭示了 MyRF特异性地激活与细胞投射、突触和细胞内相关的基因的表达 在髓鞘形成前的OL中运输。进一步地，这表明分子机制参与轴突的延长。 可能会促进OL突起的生长，这些突起与轴突接触，最终形成髓鞘。学习 属于这些类别的基因可能为研究OL分化和CNS髓鞘形成提供了新的视角。 在这方面，我们决定将重点放在Pacs2(磷酸尿酸簇分类蛋白2)上，它是 在急性Myrf基因敲除时显著下调。Pacs2是一种多功能蛋白质，它发挥着 在内质网-线粒体接触和货物分拣等不同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，包括一种综合征 自闭症的表现形式。值得注意的是，Pacs2主要由中枢神经系统中的OL表达。与RNA-Seq的发现一起 Pacs2可能是MYRF的靶标，这些观察结果表明OL可能需要Pacs2 发育和中枢神经系统髓鞘形成以及OL系细胞中的PACS2功能障碍可能是导致 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OPC的体外分化和Pacs2的表达显著下调 观察到髓鞘缺乏的自闭症小鼠模型。此外，Pacs2全身基因敲除小鼠 表现出神经学表型。目前，对Pacs2在活体OL发育中的作用知之甚少 和中枢神经系统髓鞘形成。这个项目打算通过CRE-删除OL谱系细胞中的Pacs2来解决这个问题- LoxP方法。