ER Network Shaping Mechanisms in the Hereditary Spastic Paraplegias

细胞神经科的研究集中在一些神经退行性疾病的分子机制上，包括线粒体疾病、肌张力障碍和遗传性痉挛截瘫(HSPs)。这些疾病共同困扰着数以百万计的美国人，多年来还在潜移默化地恶化，其中许多人的治疗选择有限。我们的实验室正在研究这些疾病的遗传形式，使用分子和细胞生物学方法来研究疾病基因的突变最终是如何导致细胞功能障碍的。 在这个项目中，我们将重点放在HSP上。一个主要的研究主题涉及遗传性痉挛截瘫3A(SPG3A)蛋白atlastin-1的特性和功能分析。2009年，我们在《细胞》杂志上报道，atlastin-1是一个普遍存在的GTP酶家族的成员，它与两个内质网成形蛋白家族相互作用，产生管状内质网(ER)网络。有趣的是，atlastin-1与SPG31蛋白REEP1和SPG4蛋白spastin相互作用，前者是一种内质网形成蛋白，后者是一种微管切断ATPase。2010年，我们在《临床调查杂志》上发表了一项研究，证明这三种蛋白质相互作用，与微管细胞骨架一起组织管状ER网络。由于SPG3A、SPG4和SGP31占所有HSP病例的50%以上，我们认为ER网络缺陷是HSP的主要神经病理机制。 在过去的一年里，我们建立了SPG31(基因敲除)和SPG3A(基因敲除和敲入)的动物模型，利用体内和体外研究来评估内质网形态变化的程度。我们同时使用电子显微镜和超分辨率共聚焦显微镜来检测神经元轴突内管状ER的变化，以响应这些遗传操作。此外，我们还确定了这些蛋白与另一种常染色体显性遗传HSP突变蛋白的相互作用，扩大了与内质网形成缺陷相关的HSP病例的数量。 综上所述，我们希望我们的研究将促进我们对热休克蛋白分子发病机制的理解。在分子和细胞水平上的这种理解将有望导致防止这些疾病进展的新疗法。

Research in the Cellular Neurology Unit 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 developed 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 electron microscopy 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 a protein mutated in another form of autosomal dominant HSP, expanding the number of HSP cases related to defects in ER network formation. 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.