Mouse model studies of TMEM230-linked Parkinson's disease

通过我们以前由美国帕金森病协会和NINDS资助的工作，我们已经 在20号染色体短臂上发现了一个新的遗传位点，并鉴定了一个新的PD基因TMEM 230。 TMEM 230连锁PD显示出与散发性PD一致的临床和病理特征。 TMEM 230，也称为C20 orf 30，是一个未表征的基因。我们进行了广泛的遗传和细胞 对这种新的PD基因进行体外研究。我们发现TMEM 230编码一个分泌型跨膜蛋白， 和回收囊泡，包括神经元中的突触囊泡。我们还发现TMEM 230是 路易体和路易神经突。我们进一步进行了功能研究，发现PD连锁的TMEM 230 在我们的研究中鉴定的突变体在体外损害突触囊泡运输和α-突触核蛋白清除。 TMEM 230是迄今为止在PD中鉴定的第一个突触囊泡的跨膜蛋白。我们的发现， 因此，直接指向突触囊泡功能障碍在PD发病机制中的作用。精确的函数 TMEM 230介导PD的致病机制尚不清楚。基于 TMEM 230的分子特征，及其与突触囊泡和内体标记物的关系， 在我们的研究中，我们假设TMEM 230是分泌/回收囊泡的运输蛋白，主要是 参与突触囊泡胞吐、胞吞和再循环以及神经元中的突触传递;以及 突触囊泡的功能障碍，例如受损的运输、再循环和突触传递， 这是TMEM 230相关PD以及可能的其他形式的PD的发病机制的基础。虽然我们的 体外的初步数据与该假设一致，但缺乏体内数据。适当的动物模型 尚未开发出与测试潜在致病机制相关的方法。在本申请中，我们提出 三个密切互动的具体目标，以产生相关的小鼠模型，并在体内测试这一假设。 在Specific Aim 1中，我们将开发所有三种代表性类型的小鼠模型，包括敲除、敲入和敲除。 和转基因模型。这些模型将提供独特的和必不可少的资源， 具体目标2和3。在具体目标2中，我们将测试不同类型的TMEM 230遗传效应， α-突触核蛋白清除、运动和非运动表型以及PD相关病理学的改变 小鼠模型。进一步深入了解PD表型和病理学的分子基础 TMEM 230相关突变引起的，我们将使用电生理学和细胞生物学相结合的方法， 在小鼠模型中研究突触囊泡运输和突触传递的方法 3.在小鼠模型中成功完成对这种新的PD连锁基因的拟议研究， 了解TMEM 230的生理功能和突变体的作用的基本信息 TMEM 230在PD发病机制中的作用这一应用的积极成果也可能提供一个 PD治疗开发的机制基础。

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.