Molecular mechanisms of mitochondrial dysfunction in ATP13A2-associated neurodege

描述(申请人提供)：我是一名儿科神经科医生，对神经退行性疾病有着长期的兴趣。我接受过分子遗传学方面的培训，发表了20多篇同行评议的出版物，其中10篇是第一作者。从长远来看，我致力于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。然而，我目前的技能与我作为一名独立调查员计划从事的工作之间存在着很大的差距。为了全面剖析疾病机制，我需要通过进一步的指导培训来弥合这一差距，这将使我能够发展线粒体生物学以及酵母和小鼠模型系统方面的专业知识。在我的K08中提出的研究将会做到这一点，同时也让我建立一个有意义的基础来开始我的研究生涯。对于我的K08学习，我将与大卫·皮尔斯导师合作。大卫是一位精力充沛、精力充沛的研究员，他使用酵母和小鼠模型研究神经元颈动脉脂褐质沉积症已有近20年的时间。我自己的项目专注于ATP13A2(PARK9)，一种溶酶体P5B ATPase，以及它的酵母正统ypk9(酵母PARK9)。PARK9功能丧失会导致帕金森综合症和神经元性颈动脉脂褐质沉着症。我的工作试图了解PARK9缺失导致的严重线粒体功能障碍的病因学，鉴于帕金森病和生物能量学失败之间的强烈联系，这一点尤其相关。我的初步数据显示，Ypk9p缺乏导致呼吸链复合体活性显著降低，对活性氧物种的敏感性增加，以及线粒体碎裂。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是一个重要的亚结构域，在功能上整合了内质网和线粒体之间的串扰，并参与了神经退行性变的生物学研究。为了发展我的研究技能，我将与桑福德研究公司的几位有才华的科学家一起坐在板凳上工作。这段经历将让我有机会熟练使用酵母和鼠标系统，并在最合适的情况下发挥每种模型的相对优势。我将受益于与皮尔斯博士和桑福德研究中心一群才华横溢的合作者的定期互动，他们每个人都将为我正在进行的培训提供帮助，同样重要的是，他们将为项目的成功提供帮助。此外，我还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，这代表着作为一名研究人员迅速发展的巨大机会。除了这次实践训练，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以及酵母和鼠标系统方面的专业知识，以补充我的实践经验。我还将学习领导力发展、生物统计学和计算方法等课程，以完善我的教育。我很幸运得到了我的机构Sanford Research的热情支持，该机构已经为我提供了75%的受保护的研究时间、专用空间、访问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。桑福德健康代表着上中西部地区快速增长的三级保健医疗体系，这一体系在2007年被慈善家T·丹尼·桑福德捐赠的4亿美元历史性地改变了。在过去的5年里，桑福德已经发展到在7个州的112个社区设有办事处，在爱尔兰、加纳、以色列和墨西哥设有国际办事处。Sanford Health拥有20,000名员工、34家医院和70个医学专业领域的近1000名医生，而Sanford Research已发展到包括近50个独立研究小组。Sanford Research致力于开发尖端的翻译计划，使该地区和世界各地的患者受益。皮尔斯博士致力于我的成功，我在他的指导下，在我的第一个教职年度里已经取得了巨大的进步。我很幸运地得到了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 Wemer(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，并发表了支持声明。我的具体目标是1)确定导致PARK9相关疾病线粒体失败的基本机制；2)确定PARK9在MAM中的功能；以及3)将线粒体结构和功能与ATP13A2基因敲除小鼠的疾病发生联系起来。实现这些目标将使我能够发展我所需的专业知识，以开发创新的独立研究计划，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的考虑。

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.