Role of DJ1 in mitochondrial biogenergetics and neuronal metabolism

DJ1 在线粒体生物发生学和神经元代谢中的作用

• 批准号: 10434136
• 负责人: Elizabeth Ann Jonas
• 金额: $ 47.51万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10434136
• 关键词: ATP Synthesis Pathway; Aging; Animals; Axon; Behavior; Binding; Biogenesis; Biological Assay; Cells; Complex; Corpus striatum structure; Coupling; Crista ampullaris; Cues; Distal; Dominant-Negative Mutation; Dopamine; Dopaminergic Cell; Fibroblasts; Fluorescent in Situ Hybridization; Functional disorder; Genetic Translation; Growth; Growth Factor; Knock-in; Knock-in Mouse; Knock-out; Knockout Mice; Label; Length; Ligation; Link; Measures; Membrane; Messenger RNA; Metabolic; Mitochondria; Morphology; Movement; Mus; Mutation; Neurons; Outer Mitochondrial Membrane; Oxidative Phosphorylation; PARK7 gene; Parkinson Disease; Patients; Peroxidases; Prevention; Process; Production; Protein Biosynthesis; Protein Subunits; Proteins; RNA; Readiness; Reporting; Ribosomes; Role; Signal Transduction; Site; Techniques; Viral; ascorbate; dopaminergic neuron; early onset; improved; in vivo; induced pluripotent stem cell; motor behavior; motor disorder; mutant; neuronal growth; neuronal metabolism; overexpression; preservation; prevent; protein activation; repaired; response; stoichiometry; therapy development

Abstract Jonas, EA Familial Parkinson’s disease (PD) protein DJ-1 mutations are linked to mitochondrial deficits and early onset PD but the exact mechanisms are unclear. We have found that mutations in DJ-1 and DJ-1 knock out increase mitochondrial uncoupling and hamper extension and branching of dopaminergic neuronal processes. The uncoupling is associated with abnormal ATP synthase efficiency and stoichiometry. The changes in ATP synthase are also associated with mitochondrial morphological changes including changes in cristae number, mitochondrial length and mitochondrial biogenesis. New protein synthesis is crucial to repair and maintain dopaminergic endings in the striatum and for normal dopamine release capacity. The activation of new protein synthesis is in response to cues such as growth factors or neuronal stimulation, but we hypothesize that new protein synthesis fails to occur when the stoichiometry of the ATP synthase is abnormal. In keeping with this, we find that protein synthesis levels are very low in patient cells and in DJ-1 KO mouse neurons. DJ-1 is bound to ATP synthase β subunit and ATP synthase β subunit mRNA. We therefore hypothesize that the mechanism of decreased growth and branching of DJ-1 deficient dopaminergic neurons is related to a lack of DJ-1 targeted, mitochondrially- regulated, mRNAs that are required for neuronal growth, repair and plasticity of dopaminergic cells. If we can restore the normal rate of protein synthesis downstream of DJ-1 in a spatially and temporally precise manner, we hypothesize we will slow neuronal ending degradation in DJ-1 mutant patients and preserve dopamine release capability. In keeping with this, we have now restored protein synthesis rates in patient cells by overexpression of ATP synthase β subunit. We now plan to determine if we can rescue low protein synthesis rates and neuronal growth in DJ-1 KO neurons, in neurons expressing mutant DJ-1 and in vivo in DJ-1 KO animals. We will do this by overexpression of ATP synthase β subunit by AAV (viral) delivery in neurons. In vivo, we will restore the normal coupling of mitochondria by crossing the DJ-1 KO mice with a low c-subunit leak KI mouse. We will restore the normal ATP synthase stoichiometry in vivo by crossing the DJ- 1 KO mouse with a mouse overexpressing ATP synthase β subunit. We hope to preserve neuronal endings in the striatum and prevent the onset of dysfunctional movement in the mice. Our observations suggest a connection between ATP synthase complex stoichiometry, protein synthesis rates and PD. Our plan comprises a promising new strategy to develop therapies for PD patients.

抽象的 乔纳斯，EA 家族性帕金森病 (PD) 蛋白 DJ-1 突变与线粒体缺陷有关 早发性PD，但确切机制尚不清楚。我们发现 DJ-1 和 DJ-1 敲除增加线粒体解偶联并阻碍线粒体的延伸和分支 多巴胺能神经元过程。解偶联与 ATP 合酶异常有关 效率和化学计量。 ATP合成酶的变化也与线粒体有关 形态变化包括嵴数量、线粒体长度和 线粒体生物发生。新的蛋白质合成对于修复和维持多巴胺能至关重要 纹状体的末端和正常的多巴胺释放能力。新蛋白质的激活 合成是对生长因子或神经元刺激等线索的反应，但我们 假设当 ATP 合成酶的化学计量达到一定值时，新的蛋白质合成无法发生 是不正常的。与此相一致的是，我们发现患者细胞中的蛋白质合成水平非常低 以及 DJ-1 KO 小鼠神经元。 DJ-1 与 ATP 合酶 β 亚基和 ATP 合酶 β 结合 亚基 mRNA。因此，我们假设生长和分枝减少的机制 DJ-1 缺乏的多巴胺能神经元与缺乏 DJ-1 靶向、线粒体相关 神经元生长、修复和多巴胺能可塑性所需的受调节的 mRNA 细胞。如果我们能够在空间和空间上恢复 DJ-1 下游蛋白质合成的正常速率 以时间精确的方式，我们假设我们将减缓 DJ-1 中神经元末梢的退化 突变患者并保留多巴胺释放能力。与此相一致，我们现在有 通过过度表达 ATP 合酶 β 亚基，恢复患者细胞中的蛋白质合成率。我们 现在计划确定我们是否可以挽救 DJ-1 中的低蛋白质合成率和神经元生长 KO 神经元，表达突变 DJ-1 的神经元以及 DJ-1 KO 动物体内的神经元。我们会这样做 通过 AAV（病毒）在神经元中传递，过度表达 ATP 合成酶 β 亚基。在体内，我们将 通过与低c亚基的DJ-1 KO小鼠杂交恢复线粒体的正常偶联 泄漏 KI 鼠标。我们将通过跨越 DJ- 恢复体内正常的 ATP 合酶化学计量。 1 只 KO 小鼠，小鼠过度表达 ATP 合酶 β 亚基。我们希望保留 纹状体中的神经元末梢并防止小鼠运动功能障碍的发生。 我们的观察表明 ATP 合酶复合物化学计量、蛋白质之间存在联系 合成速率和PD。我们的计划包括一项有前途的新战略，旨在开发治疗方法 帕金森病患者。