BLR&D Research Career Scientist Award Application

BLR

• 批准号: 10265408
• 负责人: Gary Cecchini
• 金额: --
• 依托单位: VETERANS AFFAIRS MED CTR SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10265408
• 关键词: Alzheimer' s Disease; Amino Acids; Apoptosis; Architecture; Area; Award; Bacterial Model; Binding; Binding Proteins; Biochemical; Biochemistry; Bioenergetics; Brain Injuries; Cardiac; Cells; Clinical; Coenzyme Q10; Complex; DNA; Development; Diabetes Mellitus; Diagnosis; Disease; Early Diagnosis; Enzymes; Epigenetic Process; Ethics; Family; Fumarates; Functional disorder; Gene Expression; Generations; Genes; Genetic Polymorphism; Genetic Transcription; Health; Heart Diseases; Heme; Homeostasis; Human; Human body; Hypoxia Inducible Factor; Inflammation; Inflammatory; Inflammatory Response; Injury; Investigation; Ischemia; Kidney Diseases; Laboratories; Laboratory Study; Lead; Lipids; Maintenance; Malignant Neoplasms; Malonates; Membrane; Methods; Minerals; Mitochondria; Mitochondrial Proteins; Modeling; Multienzyme Complexes; Multiple Sclerosis; Muscle Weakness; Mutation; Myocardial Infarction; NADH; NADH dehydrogenase (ubiquinone); Nerve Degeneration; Nerve Regeneration; Neurodegenerative Disorders; Nobel Prize; Organelles; Oxidation-Reduction; Oxidative Phosphorylation; Oxides; Paper; Parkinson Disease; Pathway interactions; Patients; Permeability; Pharmaceutical Preparations; Philosophy; Physiology; Play; Point Mutation; Poison; Process; Procollagen-Proline Dioxygenase; Protein Region; Protons; Psoriasis; Publishing; Quinone Reductases; Reactive Oxygen Species; Regulatory Element; Relapse; Reperfusion Therapy; Research; Resolution; Respiration; Respiratory Chain; Roentgen Rays; Role; Science; Scientist; Series; Severities; Signal Transduction; Signaling Molecule; Stress; Stroke; Structure; Study models; Succinate Dehydrogenase; Succinates; System; Therapeutic; Therapeutic Uses; Three-dimensional analysis; Time; Traumatic Brain Injury; Ubiquinone; United States National Academy of Sciences; Vascular remodeling; Veterans; Vitamins; Work; career; cofactor; design; healing; histone demethylase; in vivo; inhibitor/antagonist; insight; member; method development; mitochondrial dysfunction; mitochondrial membrane; mitochondrial metabolism; mouse model; news; nuclear factor-erythroid 2; nucleotide metabolism; patient population; prevent; programs; protein complex; protein metabolism; protein structure; protein structure function; research and development; respiratory; response; small molecule inhibitor; success; three dimensional structure; tumor

Our laboratory is focused on understanding how mitochondrial function contributes to health and disease. As the major energy generating organelle of the cell dysfunction of mitochondria has been implicated in debilitating diseases prevalent in the VA patient population. These include, neurodegenerative diseases (Parkinson's, Alzheimer's), diabetes, cancer, and heart disease. Altered mitochondrial metabolism can result in changed levels of tricarboxylic (TCA) cycle metabolites, (such as succinate or fumarate), that act as signaling molecules to promote a pro-inflammatory state. This can lead to changes in gene transcription, through the induction of reactive oxygen species (ROS), stabilization of hypoxia- inducible factor-1α (HIF-1α), or the nuclear factor erythroid-2-related factor-2 (Nrf2) transcription pathway that responds to pro-inflammatory stress. Our laboratory investigates the structure and function of two essential members of the mitochondrial respiratory chain both of which reduce ubiquinone (CoQ10) used by the oxidative phosphorylation system to generate energy. We study the function of Complex I (NADH:ubiquinone oxidoreductase) which is the largest membrane-bound component of the mitochondrion. NADH generated by the TCA cycle is used by Complex I to reduce CoQ10 and this activity controls the NADH/NAD+ ratio. The enzyme is regulated by a structural change near the membrane domain termed the Active/De-Active (A/D) transition, which we first showed occurred in vivo. We also study succinate dehydrogenase (SDH/Complex II) which is a membrane-bound heterotetramer of dual function. SDH oxidizes succinate to fumarate in the TCA cycle while reducing CoQ10 for energy generation. Malfunction of SDH results in accumulation of succinate in the cell which promotes inflammation. It has been shown that inhibitors of SDH can have a positive effect in treating damage form ischemia/reperfusion in both stroke and cardiac models. Our studies of SDH have shown how the reversible inhibitor malonate binds to the enzyme and causes inhibition. We are now focused on understanding how we can regulate the activity and structure of both Complexes I & II so that this information can be used to treat disease. One model we will use is to investigate how TCA cycle metabolites can be used to treat traumatic brain injury (TBI) or stroke. Dimethyl fumarate (DMF) is an approved drug for treating relapsing multiple sclerosis and psoriasis and Dimethyl malonate (DMM) is a cell-permeable non-toxic compound which in vivo can be used to inhibit SDH. We hypothesize that in the brain injury model that DMM will block succinate accumulation following injury and prevent the signaling that produces ROS during ischemia/reperfusion; thus, reducing inflammation, the severity of the injury, and enhance healing. We use mouse models for these studies. We will also determine if the epigenetic modifier DMF can reduce the inflammation caused by TBI thus lessening the severity of the injury and enhance neuro-regeneration. We were the first to determine the x-ray structure of SDH and have provided major insight into its catalytic mechanism and function. How the enzyme complex is assembled, however, remains and area of intense investigation. We are now studying the assembly of human Complex II using known human assembly factors, needed for incorporation of redox cofactors necessary for function of the enzyme. It has been shown that when assembly is compromised this can lead to tumor formation in humans. We have had success expressing and analyzing the three-dimensional structure of the human structural subunits of SDH expressed in bacterial models. Thus, for the first time the structure of these assembly intermediates will be known. This information is needed to develop small molecule inhibitors/activators that can be used for treatment of diseases associated with mitochondrial dysfunction and control metabolite levels in cells.

我们的实验室专注于了解线粒体功能如何为健康和 疾病。由于线粒体的细胞功能障碍的主要能量产生细胞器已是 在VA患者人群中普遍存在的疾病中实施。这些包括神经退行性 疾病（帕金森氏症，阿尔茨海默氏症），糖尿病，癌症和心脏病。线粒体改变 代谢可能会导致三核酸（TCA）周期代谢水平变化（例如琥珀酸酯或 富马酸盐），充当信号分子以促进促炎状态。这可能导致变化 基因转录，通过诱导活性氧（ROS），低氧稳定 诱导因子1α（HIF-1α）或核因子红系2相关因子-2（NRF2）转录途径 这对促炎压力的反应。我们的实验室调查了两个的结构和功能 线粒体呼吸链的基本成员两者都减少了泛氨基酮（COQ10） 氧化物磷酸化系统产生能量。我们研究复合物I的功能 （NADH：泛氨基氧化激酶），这是线粒体最大的膜结合成分。 由TCA周期产生的NADH由复合物I使用来减少COQ10，并且该活动控制 NADH/NAD+比率。该酶受到称为膜结构域附近的结构变化的调节 我们首先在体内发生的主动/去激活（A/D）过渡。我们还研究琥珀酸酯 脱氢酶（SDH/复合物II）是双重功能的膜结合的异驱动器。 SDH 在TCA循环中氧化琥珀酸酯至富马酸盐，同时减少COQ10的能量产生。故障 SDH导致琥珀酸在促进注射的细胞中积累。已经表明 SDH的抑制剂在治疗损伤形式中具有积极作用 心脏模型。我们对SDH的研究表明，可逆抑制剂疟疾如何与酶结合 并引起抑制作用。现在，我们专注于了解如何调节活动和 两种复合物I＆II的结构，因此可以使用此信息来治疗疾病。 我们将使用的一种模型是研究如何使用TCA循环代谢物治疗创伤 脑损伤（TBI）或中风。富马酸二甲基二甲基（DMF）是一种批准的药物，用于治疗复发多重 硬化症和牛皮癣和丙二酯（DMM）是一种可渗透的无毒化合物，它在 体内可用于抑制SDH。我们假设在脑损伤模型中DMM会阻塞 受伤后的琥珀酸盐积累，并防止在期间产生ROS的信号传导 缺血/再灌注；因此，减少感染，损伤的严重程度并增强愈合。我们使用 这些研究的小鼠模型。我们还将确定表观遗传修饰符DMF是否可以减少 TBI引起的炎症因此减轻了损伤的严重程度和增强的神经再生。 我们是第一个确定SDH X射线结构的人，并为其提供了重大见解 催化机制和功能。但是，如何组装酶复合物以及 激烈的投资。我们现在正在使用已知人类研究人类复合物II的组装 装配因子，用于酶功能所需的氧化还原辅助因子所需的因素。它有 我们已经妥协了组装时，这会导致人类的肿瘤形成。我们有 成功表达和分析SDH人类结构亚基的三维结构的成功 在细菌模型中表达。那是这些组装中间体的结构首次是 已知。需要此信息来开发可用于的小分子抑制剂/激活剂 与细胞中线粒体功能障碍和控制代谢物水平相关的疾病的治疗。