Molecular mechanisms of mitochondrial dysfunction in ATP13A2-associated neurodege

描述（由申请人提供）：我是一名儿科神经科医生，长期对神经退行性疾病感兴趣。我接受过分子遗传学方面的培训，发表了20多篇同行评审的论文，其中10篇是第一作者。从长远来看，我承诺尽一切可能为我的神经退行性疾病患者带来新的治疗方法，无论这些治疗方法来自我的实验室还是同事的实验室。然而，我目前的技能和我计划作为一名独立调查员从事的工作之间存在着巨大的差距。为了全面剖析疾病的机制，我需要通过进一步的指导培训来弥补这一差距，这将使我能够在线粒体生物学、酵母和小鼠模型系统方面发展专业知识。在我的K08中提出的研究将做到这一点，同时也让我为开始我的研究生涯奠定一个有意义的基础。在K08的学习中，我将与导师David Pearce合作。David是一位充满活力和活力的研究者，他使用酵母和小鼠模型研究了近20年的神经颈动脉脂褐质病。我自己的项目重点是ATP13A2 (PARK9)，一种溶酶体P5B atp酶，以及它的酵母正统ypk9（酵母PARK9）。PARK9功能丧失可导致帕金森病和神经性颈动脉脂褐变。我的工作旨在了解与PARK9丢失一起发生的严重线粒体功能障碍的病因学，这尤其与帕金森病和生物能量衰竭之间的紧密联系有关。我的初步数据表明，Ypk9p缺乏导致呼吸链复合物活性显著降低，对活性氧的敏感性增加，以及线粒体断裂。这些发现与a-向斜的影响无关。此外，我们最近认识到PARK9也存在于酵母和哺乳动物细胞的线粒体相关ER膜（MAM）中。MAM是一个重要的子结构域，它在功能上整合内质网和线粒体之间的串扰，并与神经变性的生物学有关。为了提高我的研究技能，我将与桑福德研究所的几位才华横溢的科学家一起在板凳上工作。这次经历将使我有机会熟练使用酵母和小鼠系统，并在最合适的环境中利用每种模型的相对优势。我将受益于与Pearce博士和桑福德研究中心一群才华横溢的合作伙伴的定期互动，他们每个人都将提供他们的专业知识，以促进我正在进行的培训，同样重要的是，项目的成功。此外，我将前往约翰霍普金斯大学，在那里我将在Hiromi Sesakis实验室接受培训。Hiromi是酵母和小鼠系统线粒体动力学方面的专家，这是一个巨大的机会，可以迅速发展为一名研究者。除了这个实践培训，我还会参加几个短期课程。这些课程将帮助我发展线粒体生物学、酵母和小鼠系统方面的专业知识，以补充我的实践经验。我还将参加领导力发展、生物统计学和计算方法方面的课程，以完善我的教育。我很幸运地得到了我所在的机构桑福德研究所的热情支持，它已经为我提供了75%的研究保护时间、专用空间、使用核心和所需设备的机会，以及帮助我建立研究计划的机构津贴。桑福德健康代表了中西部上游地区快速增长的三级保健医疗系统，该系统由2007年慈善家T. Denny Sanford提供的历史性4亿美元礼物改造而成。在过去的5年里，桑福德已经发展到包括七个州的112个社区，在爱尔兰，加纳，以色列和墨西哥设有国际办事处。桑福德健康拥有20,000名员工，34家医院，在70个医学专业领域拥有近1000名医生，而桑福德研究已经发展到包括近50个独立的研究小组。桑福德研究中心致力于开发尖端的转化项目，使整个地区和世界的患者受益。皮尔斯博士致力于我的成功，在他作为我的导师的第一年里，我已经取得了巨大的进步。我很幸运能得到许多有才华的同事和合作者的支持。这包括Sergio Padilla-Lopez（酵母遗传学和生物化学，线粒体生物学），Pete Vitiello（氧化还原生物学），Jill Weimer（小鼠神经变性模型），Keith Miskimins（线粒体动力学和疾病功能）和Attila Kovacs（小鼠表型和原代神经元培养）。所有人都热情地支持我的申请，并提供了支持声明。我的具体目标是：1)确定导致park9相关疾病中线粒体衰竭的基本机制；2)确定PARK9在MAM中的功能；3)在ATP13A2基因敲除小鼠中，线粒体结构和功能与疾病的出现相关。实现这些目标将使我能够发展我需要的专业知识，以开发一个创新的独立研究项目，同时为后续的治疗研究奠定基础。我很感激能获得这个奖项。

DESCRIPTION (provided by applicant): I am a pediatric neurologist with a long-standing interest in neurodegenerative disease. I have been trained in molecular genetics, and have published over 20 peer-reviewed publications, 10 as first author. In the long-term, I am committed to do everything possible to bring new treatments to my patients with neurodegenerative disorders, whether these therapies come from my lab or that of a colleague. However, a significant gap exists between my present skill set and the work I plan to pursue as an independent investigator. In order to fully dissect mechanisms of disease, I will need to bridge this gap with furthered mentored training that will allow me to develop expertise in mitochondrial biology and yeast and mouse model systems. The studies proposed within my K08 will do just that, while also allowing me to build a meaningful foundation to launch my research career. For my K08 studies, I will partner with mentor David Pearce. David is an energetic and dynamic investigator who has studied neuronal carotid lipofuscinosis using yeast and mouse models for almost 20 years. My own project focuses on ATP13A2 (PARK9), a lysosomal P5B ATPase, and its yeast orthodox ypk9 (yeast PARK9). PARK9 loss of function leads to both Parkinsonism and neuronal carotid lipofuscinosis. My work seeks to understand the etiology of the profound mitochondrial dysfunction that occurs with PARK9 loss, which is particularly relevant given the strong link between Parkinsonism and bioenergetics failure. My preliminary data indicates that Ypk9p deficiency leads to markedly diminished respiratory chain complex activity and increased sensitivity to reactive oxygen species, as well as mitochondrial fragmentation. These findings are independent of the effects of a-syncline. In addition, we have newly recognized that PARK9 is also found within the mitochondria-associated ER membrane (MAM) in both yeast and mammalian cells. The MAM is an important subdomain that functionally integrates cross-talk between the ER and mitochondria, and has been implicated in the biology of neurodegeneration. In order to develop my research skills, I will work at the bench alongside several talented scientists at Sanford Research. This experience will afford me the opportunity to become adept with both yeast and mouse systems, and to exploit the relative strengths of each model in the most appropriate contexts. I will have the benefit of regular interactions with Dr. Pearce and a talented cadre of collaborators at Sanford Research, each of whom will lend their expertise to facilitate my ongoing training, and equally importantly, the project's success. In addition, I will travel to Johns Hopkins, where I will train in Hiromi Sesakis lab. Hiromi is an expert in mitochondrial dynamics in both yeast and murine systems, and this represents a tremendous opportunity to rapidly develop as an investigator. In addition to this hands-on training, I will take several short courses. These courses will help me to develop expertise in mitochondrial biology and yeast and mouse systems to complement my practical experience. I will also take courses in leadership development and biostatistics and computational methods to round out my education. I am fortunate to have enthusiastic support from my institution, Sanford Research, which has already provided me with 75% protected time for research, dedicated space, access to cores and needed equipment, and an institutional allowance to help establish my research program. Sanford Health represents a rapidly growing tertiary care medical system within the Upper Midwest which was transformed by a historic $400 million gift in 2007 from philanthropist T. Denny Sanford. In the last 5 years, Sanford has grown to include locations in 112 communities in seven states with international sites in Ireland, Ghana, Israel and Mexico. Sanford Health encompasses 20,000 employees, 34 hospitals, and nearly 1000 physicians in 70 specialty areas of medicine, while Sanford Research has grown to include nearly 50 independent research groups. Sanford Research is committed to developing cutting-edge translational programs that will benefit patients throughout the region and the world. Dr. Pearce is committed to my success, and I have already grown tremendously during my first faculty year with him as my mentor. I am fortunate to have the support of a number of talented colleagues and collaborators. This includes Sergio Padilla-Lopez (yeast genetics and biochemistry, mitochondrial biology), Pete Vitiello (redox biology), Jill Weimer (mouse models of neurodegeneration), Keith Miskimins (mitochondrial dynamics and function in disease), and Attila Kovacs (mouse phenotyping and primary neuronal culture). All enthusiastically support my application, and have contributed Statements of Support. My specific aims seek to 1) identify the fundamental mechanisms that lead to mitochondrial failure in PARK9-associated disease; 2) determine the function of PARK9 within the MAM; and 3) correlate mitochondrial structure and function with the emergence of disease in an ATP13A2 knockout mouse. Accomplishing these aims will allow me to develop the expertise I need to develop an innovative independent research program while laying the foundation for subsequent treatment studies. I am grateful to be considered for this award.