Endocytic Mechanisms in the Hereditary Spastic Paraplegias

遗传性痉挛性截瘫的内吞机制

基本信息

批准号：
8746852
负责人：
Craig Blackstone
金额：
$ 142.97万
依托单位：
NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
依托单位国家：
美国
项目类别：
财政年份：
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/8746852
关键词：
Actins Adaptor Signaling Protein American Amyotrophic Lateral Sclerosis Cells Cellular biology Clinical Cytokinesis Cytoskeleton Defect Disease Disease model Dystonia Endosomes Fibroblasts Functional disorder Genes Genetic Goals Hereditary Spastic Paraplegia Human In Situ Inherited Investigation Journals Knockout Mice Laboratories Lead Length Maps Mediating Membrane Protein Traffic Mitochondrial Diseases Modeling Molecular Molecular Biology Molecular Genetics Motor Neurons Mus Mutate Mutation Nervous system structure Neurodegenerative Disorders Neurology Neurons Pathogenesis Pathway interactions Patients Proteins Publishing Reporting Research Shapes Small Interfering RNA Systems Analysis Troyer syndrome axonopathy clinically relevant functional group hereditary neuropathy induced pluripotent stem cell insight mouse model nervous system disorder novel prevent protein complex protein function structural biology trafficking

项目摘要

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 the current project, we have been investigating HSPs that result from defects in proteins implicated in endocytic trafficking. These include the complicated HSP known as Troyer syndrome (SPG20), which is cause by mutations in the spartin gene that likely result in complete loss of the spartin protein. We have recently reported that the spartin protein interacts with the ESCRT-III protein IST1 and is involved in cytokinesis. We are currently investigating the function of spartin in the nervous system by analyzing spartin-null mice that we have generated as a murine model of Troyer syndrome. A detailed characterization of this mouse was published in the journal Human Molecular Genetics in 2012. Over the past year, we have begun to study the interplay of the proteins that are mutated in SPG11 and SPG15. These proteins interact with one another as well as with a new adaptor protein complex -- AP5. We are using siRNA-mediated depletion studies and structural approaches to investigate the functions of these proteins in cells. Lastly, we are investigating the functions of the SPG8 protein strumpellin, which is part of the WASH protein complex implicated in the shaping of endosomes through alterations of the actin cytoskeleton. For all of these disorders, we have patient-derived fibroblasts from which we are creating induced pluripotent stem cells, and subsequently telencephalic neurons to model these disorders in situ. 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.

细胞神经病学部门的研究重点是许多神经退行性疾病的分子机制，包括线粒体疾病，肌张力障碍和遗传性痉挛性跨性疾病（HSPS）。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。在当前的项目中，我们一直在研究由涉及内吞贩运的蛋白质缺陷导致的HSP。其中包括复杂的HSP称为Troyer综合征（SPG20），这是由于Spartin基因中的突变引起的，可能会导致Spartin蛋白完全丧失。我们最近报道说，斯巴丁蛋白与ESCRT-III蛋白IST1相互作用，并参与细胞因子。我们目前正在通过分析我们作为Troyer综合征的鼠模型产生的Spartin-null小鼠来研究Spartin在神经系统中的功能。该小鼠的详细表征发表在2012年的《人类分子遗传学》杂志上。在过去的一年中，我们开始研究SPG11和SPG15中突变的蛋白质的相互作用。这些蛋白质彼此相互作用，并与新的衔接蛋白复合物-AP5相互作用。我们正在使用siRNA介导的耗竭研究和结构方法来研究这些蛋白在细胞中的功能。最后，我们正在研究SPG8蛋白strumpellin的功能，该功能是通过肌动蛋白细胞骨架的改变与内体形成有关的洗涤蛋白复合物的一部分。对于所有这些疾病，我们都有患者衍生的成纤维细胞，我们从中产生了诱导的多能干细胞，然后端切脑神经元在原位对这些疾病进行建模。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。