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，发表过20多篇同行评议的论文，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，无论这些疗法来自我的实验室还是同事的实验室。然而，我目前的技能与我作为独立调查员计划从事的工作之间存在着重大差距。为了充分剖析疾病的机制，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和酵母和小鼠模型系统方面的专业知识。在我的K 08中提出的研究将做到这一点，同时也让我建立一个有意义的基础，开始我的研究生涯。 对于我的K 08学习，我将与导师大卫皮尔斯合作。大卫是一个精力充沛和充满活力的研究者，他使用酵母和小鼠模型研究神经元颈动脉脂褐质沉积症近20年。我自己的项目集中在ATP 13 A2（PARK 9），一种溶酶体P5 B ATP酶，及其酵母正统ypk 9（酵母PARK 9）。PARK 9功能丧失导致帕金森综合征和神经元颈动脉脂褐质沉积症。我的工作旨在了解PARK 9丢失引起的线粒体功能障碍的病因，鉴于帕金森症和生物能量学失败之间的密切联系，这一点特别重要。我的初步数据表明，YPK 9 p缺乏导致呼吸链复合体活性显著降低，对活性氧的敏感性增加，以及线粒体碎片化。这些发现是独立的a-向斜的影响。此外，我们新发现PARK 9也存在于酵母和哺乳动物细胞中的Era相关ER膜（MAM）内。MAM是一个重要的亚结构域，在功能上整合了ER和线粒体之间的串扰，并与神经退行性疾病的生物学有关。 为了发展我的研究技能，我将与桑福德研究所的几位才华横溢的科学家一起工作。这一经验将使我有机会熟练使用酵母和小鼠系统，并在最适当的情况下利用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所一群才华横溢的合作者的定期互动，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，该项目的成功。此外，我将前往约翰霍普金斯，在那里我将在Hiromi Sesakis实验室进行培训。Hiromi是酵母和小鼠系统中线粒体动力学的专家，这代表了作为研究人员快速发展的巨大机会。 除了这个实践培训，我将参加几个短期课程。这些课程将帮助我发展线粒体生物学和酵母和小鼠系统的专业知识，以补充我的实践经验。我还将参加领导力发展和生物统计学和计算方法的课程，以完善我的教育。 我很幸运得到了我所在机构桑福德研究所的热情支持，它已经为我提供了75%的研究时间，专用空间，获得核心和所需设备，以及帮助建立我的研究计划的机构津贴。桑福德健康代表了上中西部迅速发展的三级保健医疗系统，2007年慈善家T。丹尼·桑福德。在过去的5年里，桑福德已经发展到包括在爱尔兰，加纳，以色列和墨西哥的国际网站在7个州的112个社区的位置。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，我已经在我的第一个教师年与他作为我的导师极大地成长。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（神经变性的小鼠模型），基思Miskimins（疾病中的线粒体动力学和功能）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PARK 9相关疾病中线粒体功能衰竭的基本机制; 2）确定PARK 9在MAM中的功能; 3）将线粒体结构和功能与ATP 13 A2敲除小鼠中疾病的出现相关联。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计划，同时为后续的治疗研究奠定基础。我很感激被考虑这个奖项。

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.