ER Network Shaping Mechanisms in the Hereditary Spastic Paraplegias

遗传性痉挛性截瘫的 ER 网络塑造机制

• 批准号: 8746791
• 负责人: Craig Blackstone
• 金额: $ 102.12万
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8746791
• 关键词: ATP phosphohydrolase; Accounting; American; Amyotrophic Lateral Sclerosis; Animal Model; Area; Axon; Cells; Cellular biology; Clinical; Clinical Trials; Collaborations; Confocal Microscopy; Cytoskeleton; Defect; Disease; Dystonia; Electron Microscopy; Endoplasmic Reticulum; Family; Functional disorder; Genes; Genetic; Goals; Guanosine Triphosphate Phosphohydrolases; Hereditary Spastic Paraplegia; Human; In Situ; Inherited; Investigation; Journals; Knock-in Mouse; Knock-out; Laboratories; Lead; Length; Lipids; Maps; Membrane Protein Traffic; Microtubules; Mitochondrial Diseases; Modeling; Molecular; Molecular Biology; Morphology; Motor Neurons; Mutate; Mutation; Neurodegenerative Disorders; Neurons; Pathogenesis; Pathway interactions; Patients; Production; Proteins; Publishing; Reporting; Research; Resolution; Shapes; Spastic Paraplegia; Tubular formation; axonopathy; clinically relevant; functional group; genetic manipulation; hereditary neuropathy; in vivo; induced pluripotent stem cell; insight; member; mouse model; nervous system disorder; neurogenetics; novel; prevent; response; spastin; structural biology

Research in the Cell Biology Section, Neurogenetics Branch focuses on the molecular mechanisms underlying a number of neurodegenerative disorders, including mitochondrial disorders, dystonia, and the hereditary spastic paraplegias (HSPs). These disorders, which together afflict millions of Americans, worsen insidiously over a number of years, and treatment options are limited for many of them. Our laboratory is investigating inherited forms of these disorders, using molecular and cell biology approaches to study how mutations in disease genes ultimately result in cellular dysfunction. In this project, we are focusing on the HSPs. One major research theme involves the characterization and functional analysis of the hereditary spastic paraplegia type 3A (SPG3A) protein, atlastin-1. In 2009, we reported in the journal Cell that atlastin-1 is a member of a ubiquitous family of GTPases that interact with two families of ER shaping proteins to generate the tubular endoplasmic reticulum (ER) network. Interestingly, atlastin-1 interacts with the SPG31 protein REEP1, which is an ER shaping protein, as well as the SPG4 protein spastin, a microtubule-severing ATPase. In 2010, we published a study in the Journal of Clinical Investigation demonstrating that these three proteins interact with one another to organize the tubular ER network in conjunction with the microtubule cytoskeleton. Since SPG3A, SPG4, and SGP31 account for well over 50% of all HSP cases, we suggest ER network defects as the predominant neuropathologic mechanism for the HSPs. Over the past year, we have continued to develop animal models for SPG31 (knock out) and SPG3A (knockout and knock in) to evaluate the extent of ER morphology changes using both in vivo and ex vivo studies. We are employing both high-throughput electron microscopy (in collaboration with Dr. Mark Terasaki) and super-resolution confocal microscopy to examine the changes in tubular ER within neuronal axons in response to these genetic manipulations. In addition, we have identified interactions of these proteins with several other proteins mutated in the HSPs, expanding the number of HSP cases related to defects in ER network formation. A key aspect of ER function possibly related to disease pathogenesis is the formation of lipid droplets, and this is an area of emphasis for our cellular and organismal studies. Last, we are actively generating in situ models for many of the HSPs through the production of patient-derived, induced pluripotent stem cells that are then differentiated into telecephalic neurons, in collaboration with Dr. Xue-Jun Li. Taken together, we expect that our studies will advance our understanding of the molecular pathogenesis of the HSPs. Such an understanding at the molecular and cellular levels will hopefully lead to novel treatments to prevent the progression of these disorders.

在细胞生物学部分的研究，神经遗传学分支集中在许多神经退行性疾病的基础机制上，包括线粒体疾病，肌张力障碍和遗传性痉挛性瘫痪（HSP）。 这些疾病共同遭受了数百万美国人的困扰，在多年中，阴险的恶化，其中许多疾病受到限制。 我们的实验室正在使用分子和细胞生物学方法研究这些疾病的遗传形式，以研究疾病基因中的突变如何最终导致细胞功能障碍。 在这个项目中，我们专注于HSP。一个主要的研究主题涉及遗传性痉挛性截瘫3A（SPG3A）蛋白Atlastin-1的表征和功能分析。 2009年，我们在Cell杂志上报道了Atlastin-1是无处不在的GTPases家族的成员，它们与两个ER塑造蛋白质的家族相互作用以产生管状内质网（ER）网络。 有趣的是，Atlastin-1与SPG31蛋白REEP1相互作用，该蛋白REEP1是一种ER成型蛋白，以及SPG4蛋白Spastin（一种微管 - 渗透性ATPase）。 2010年，我们在《临床研究杂志》上发表了一项研究，表明这三种蛋白质相互作用，以与微管细胞骨架结合组织管状ER网络。 由于SPG3A，SPG4和SGP31占所有HSP病例的50％以上，因此我们认为ER网络缺陷是HSP的主要神经病理机制。 在过去的一年中，我们继续开发用于SPG31（敲除）和SPG3A（敲除和敲门）的动物模型，以评估使用体内和离体研究的ER形态变化的程度。 我们正在使用高通量电子显微镜（与Mark Terasaki博士合作）和超分辨率共聚焦显微镜，以检查神经元轴突内的管状ER的变化，以应对这些遗传操作。 此外，我们已经确定了这些蛋白质与HSP中突变的其他几种蛋白质的相互作用，从而扩大了与ER网络形成中缺陷相关的HSP病例的数量。 ER功能可能与疾病发病机理有关的一个关键方面是脂质液滴的形成，这是我们的细胞和有机研究的重点。 最后，我们通过与Xue-jun li博士的合作，通过生产患者诱导的多能干细胞来积极地为许多HSP生成原位模型，然后将其分化为电信神经元。 综上所述，我们期望我们的研究将提高我们对HSP分子发病机理的理解。 对分子和细胞水平的这种理解将有望导致新的治疗方法，以防止这些疾病的发展。