Mouse model studies of TMEM230-linked Parkinson's disease

TMEM230相关帕金森病的小鼠模型研究

• 批准号: 9751106
• 负责人: Han-Xiang Deng
• 金额: $ 44.28万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751106
• 关键词: 20p; Affect; American; Animal Model; Biochemical; Biological; Cells; China; Chromosomes, Human, Pair 20; Clinical; Complex; DNA; Data; Defect; Dideoxy Chain Termination DNA Sequencing; Disease; Electrophysiology (science); Endocytosis; Ethnic group; Etiology; European; Exocytosis; Exploratory/Developmental Grant; Family; Functional disorder; Funding; Genes; Genetic; Genetic study; Human; Impairment; In Vitro; Individual; Integral Membrane Protein; Knock-in; Knock-out; Lewy Bodies; Link; Mediating; Missense Mutation; Modeling; Modification; Molecular; Motor; Mus; Mutation; National Institute of Neurological Disorders and Stroke; Neurites; Neurodegenerative Disorders; Neurons; Outcome; Parkinson Disease; Pathogenesis; Pathogenicity; Pathologic; Pathology; Patients; Pattern; Phenotype; Physiological; Recycling; Resources; Role; Sampling; Study models; Symptoms; Synaptic Transmission; Synaptic Vesicles; Testing; Transgenic Mice; Transgenic Model; Transgenic Organisms; Vesicle; Work; age related; alpha synuclein; base; design; disease mechanisms study; disease phenotype; effective therapy; exome sequencing; genome-wide linkage; in vivo; indexing; insight; member; mouse development; mouse model; mutant; novel; protein transport; secretory protein; therapeutic development; trafficking

Through our previous work funded by the American Parkinson Disease Association and NINDS, we have discovered a new genetic locus on the short arm of chromosome 20, and identified a new PD gene, TMEM230. TMEM230-linked PD shows clinical and pathological features compatible with those in sporadic PD. TMEM230, also known as C20orf30, is an uncharacterized gene. We performed extensive genetic and cellular studies on this new PD gene in vitro. We found that TMEM230 encodes a transmembrane protein of secretory and recycling vesicles, including synaptic vesicles in neurons. We also found that TMEM230 is a component of Lewy bodies and Lewy neurites. We further performed functional studies and found that PD-linked TMEM230 mutants identified in our study impair synaptic vesicle trafficking and alpha-synuclein clearance in vitro. TMEM230 is the first transmembrane protein of synaptic vesicles identified in PD to date. Our findings, therefore, directly point to the dysfunction of synaptic vesicles in the pathogenesis of PD. The precise functions of TMEM230, and pathogenic mechanism of the TMEM230-mediated PD remain unclear. Based on the molecular features of TMEM230, and its relationship with synaptic vesicle and endosomal markers tested in our study, we hypothesize that TMEM230 is a trafficking protein of secretory/recycling vesicles, primarily involved in synaptic vesicle exocytosis, endocytosis and recycling, and synaptic transmission in neurons; and dysfunction of the synaptic vesicles, such as impaired trafficking, recycling and synaptic transmission, underlies the pathogenesis of the TMEM230-linked PD, and possibly other forms of PD as well. Although our preliminary data in vitro are consistent with this hypothesis, in vivo data are lacking. Appropriate animal models relevant to testing potential pathogenic mechanisms have not been developed. In this application, we propose three closely interactive specific aims to generate relevant mouse models and test this hypothesis in vivo. In Specific Aim1, we will develop all three representative types of mouse models, including knockout, knockin and transgenic models. These models will provide unique and essential resources for mechanistic studies in Specific Aims 2 and 3. In Specific Aim 2, we will test the effects of different types of TMEM230 genetic modifications on alpha-synuclein clearance, motor and non-motor phenotypes, and PD-related pathology in the mouse models. To further provide insights into the molecular underpinning of PD phenotype and pathology caused by TMEM230-related mutations, we will use a combination of electrophysiological and cell biological approaches to study synaptic vesicle trafficking and synaptic transmission in the mouse models in Specific Aim 3. Successful completion of the proposed studies of this new PD-linked gene in mouse models should provide essential information for understanding the physiological functions of TMEM230, and the role of the mutant TMEM230 in the pathogenesis underlying PD. The positive outcomes from this application may also provide a mechanistic basis for PD therapeutic development.

通过我们以前由美国帕金森氏病协会和Ninds资助的工作，我们有 在20号染色体的短臂上发现了一个新的遗传基因座，并确定了一个新的PD基因TMEM230。 TMEM230连接的PD显示了与零星PD中兼容的临床和病理特征。 TMEM230，也称为C20ORF30，是一个未表征的基因。我们进行了广泛的遗传和细胞 在体外研究了该新的PD基因。我们发现TMEM230编码分泌的跨膜蛋白 和回收囊泡，包括神经元中的突触囊泡。我们还发现TMEM230是 路易的身体和路易神经突。我们进一步进行了功能研究，发现PD连接的TMEM230 在我们的研究中鉴定出的突变体会损害体外突触囊泡运输和α-核蛋白清除率。 TMEM230是迄今为止在PD中鉴定出的突触囊泡的第一个跨膜蛋白。我们的发现， 因此，直接指向PD发病机理中突触囊泡的功能障碍。精确的功能 TMEM230和TMEM230介导的PD的致病机理尚不清楚。基于 TMEM230的分子特征及其与突触囊泡和内体标记的关系 我们的研究，我们假设TMEM230是分泌/回收囊泡的运输蛋白 参与神经元的突触囊泡胞吐作用，内吞作用和回收以及突触传播；和 突触囊泡功能障碍，例如运输受损，回收和突触传播， 构成了TMEM230连接PD的发病机理，也可能是其他形式的PD。虽然我们的 体外初步数据与该假设一致，缺乏体内数据。适当的动物模型 与测试潜在的致病机制有关。在此应用程序中，我们建议 三个紧密互动的特定目的是生成相关的小鼠模型并在体内检验该假设。 在特定的AIM1中，我们将开发所有三种代表性的鼠标模型，包括敲除，敲除 和转基因模型。这些模型将为机械研究提供独特的基本资源 具体目的2和3。在特定目标2中，我们将测试不同类型的TMEM230遗传的影响 对α-突触核蛋白清除，运动和非运动表型以及与PD相关的病理的修改 鼠标模型。为了进一步提供有关PD表型和病理分子基础的见解 由TMEM230相关的突变引起，我们将使用电生理和细胞生物学的组合 在特定目标中研究小鼠模型中研究突触囊泡运输和突触传播的方法 3。在小鼠模型中成功完成该新PD连接基因的拟议研究应提供 了解TMEM230的生理功能的基本信息，以及突变体的作用 TMEM230在PD的发病机理中。该应用程序的积极结果也可能提供 PD治疗开发的机械基础。