Neurodegenerative consequences of PanK2 mutations

描述（由申请人提供）：候选人是医学博士/博士神经学家，目前是神经退行性疾病研究中心的实习生。他的目标是发展额外的研究技能和经验，成为一名独立的临床科学家，致力于了解神经退行性疾病的发病机制。拟议的研究项目侧重于神经退行性疾病与脑铁积累（NBIA），这会导致语言，运动和认知的进行性障碍。在神经病理学水平上，NBIA的特征在于铁积累、包涵体形成、氧化应激的迹象和多个神经元群体的死亡。这些特征在其他神经退行性疾病中也有不同程度的表现，包括帕金森病和阿尔茨海默病。最近在NBIA病例的一个子集中发现了泛酸激酶2（PanK 2）基因突变。PanK 2基因编码参与辅酶A（CoA）合成的酶，辅酶A合成是与许多细胞过程相关的关键途径，包括脂肪酸合成、能量产生以及可能的抗氧化剂分子的合成。该项目的长期目标是了解PanK 2突变如何导致铁积累，氧化应激，包涵体形成和神经元死亡。将表征突变体Pank 2蛋白的蛋白水解加工、线粒体定位和体外催化性质，并与野生型人PanK 2蛋白进行比较。将建立细胞培养系统，其中PanK 2表达被消除，并且其中野生型或突变PanK 2蛋白过表达。还将产生缺乏PanK 2表达的小鼠。将检查缺乏PanK 2表达的细胞系和小鼠的生化中间体水平的变化，假设其依赖于PanK 2功能。最后，将检查缺乏PanK 2的神经元和非神经元细胞的氧化应激增加、对氧化损伤的易感性、放射性标记铁的细胞和线粒体输入以及包涵体形成的迹象。

DESCRIPTION (provided by applicant): The candidate is an M.D./Ph.D neurologist who is currently a trainee in the Center for Neurodegenerative Disease Research. His goal is to develop additional research skills and experience needed to become an independent clinician scientist working to understand the pathogenesis of neurodegenerative diseases. The proposed research project focuses on neurodegeneration with brain iron accumulation (NBIA), which causes progressive impairment of speech, movement and cognition. At the neuropathological level, NBIA is characterized by iron accumulation, inclusion formation, signs of oxidative stress, and death of multiple neuronal populations. These features are also seen to varying degrees in other neurodegenerative diseases, including Parkinson's disease and Alzheimer's disease. Mutations in the gene for pantothenate kinase 2 (PanK2) were recently identified in a subset of NBIA cases. The PanK2 gene encodes an enzyme involved in coenzyme A (CoA) synthesis, a critical pathway linked to a number of cellular processes, including fatty acid synthesis, energy production, and possibly, synthesis of anti-oxidant molecules. The long term objectives of this project are to understand how PanK2 mutations lead to iron accumulation, oxidative stress, inclusion formation, and neuronal death. The proteolytic processing, mitochondrial localization and in vitro catalytic properties will by characterized for mutant Pank2 proteins and compared to the wild type human PanK2 protein. Cell culture systems will be established in which PanK2 expression is eliminated and in which wild type or mutant PanK2 proteins are over-expressed. Mice that lack PanK2 expression will also be generated. Cell lines and mice lacking PanK2 expression will be examined for changes in levels of biochemical intermediates hypothesized to be dependent on PanK2 function. Finally, neuronal and non-neuronal cells lacking PanK2 will be examined for signs of increased oxidative stress, susceptibility to oxidative injury, cellular and mitochondrial import of radio labeled iron, and inclusion formation.