Regulation of Dendrite Homeostasis by PINK1 and PKA in Models of Parkinson's Disease

帕金森病模型中 PINK1 和 PKA 对树突稳态的调节

• 批准号: 10263704
• 负责人: RUBEN K DAGDA
• 金额: $ 5万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 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 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10263704
• 关键词: A kinase anchoring protein; Adaptor Signaling Protein; Affect; Area; Binding; Biochemical; Biological Assay; Brain; Brain-Derived Neurotrophic Factor; Chemical Models; Chemicals; Cleaved cell; Cyclic AMP-Dependent Protein Kinases; Cytosol; Data; Dendrites; Disease model; Etiology; Exhibits; Fluorescence Resonance Energy Transfer; Genetic; Health; Homeostasis; Image; Impairment; In Vitro; Knock-out; Knowledge; LRRK2 gene; Lead; Length; Link; Mediating; Midbrain structure; Mitochondria; Mitochondrial Matrix; Modeling; Molecular; Molecular Biology; Molecular Weight; Mus; Mutation; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neurons; Neurotrophic Tyrosine Kinase Receptor Type 2; Oxidative Stress; PTEN gene; Parkinson Disease; Pathology; Pharmacology; Phosphorylation; Phosphorylation Site; Phosphotransferases; Process; Publishing; Regulation; Research; Research Project Grants; Role; Signal Pathway; Signal Transduction; Substantia nigra structure; Testing; Therapeutic; Western Blotting; Work; base; dopaminergic neuron; enhancing factor; experimental study; human disease; imaging approach; in vivo; innovation; motor disorder; neuronal survival; novel; phosphoproteomics; scaffold; therapy development; trafficking

PROJECT SUMMARY Mutations in PTEN-induced kinase 1 (PINK1) are associated with autosomal recessive forms of Parkinson's disease (PD). In the mitochondrion, full-length PINK1 is proteolytically processed to lower molecular weight forms which are exported to the cytosol. While full-length PINK1 has been implicated in regulating mitophagy and mitochondrial function, the understanding of the functional role(s) of endogenous cleaved PINK1 (c- PINK1) in the brain is limited. Our recent research demonstrates a link of c-PINK1 and regulation of mitochondrial trafficking and dendrite outgrowth via PKA. This model is supported by preliminary and published data from our group, showing that loss of PINK1 function in vivo and in vitro impairs PKA-mediated dendrite connectivity and mitochondrial trafficking, but the connections between these two mechanisms are unresolved. The studies proposed will fill a critical void in our understanding of how c-PINK1 regulates PKA signaling to enhance dendrite connectivity and mitochondrial trafficking in PD models. In Aim 1, the molecular mechanisms by which PINK1 and dendrite-localized PKA protect dendrites from oxidative stress will be elucidated using image-based and molecular biology approaches. Aim 2 will elucidate the mechanisms by which PINK1 and PKA regulate mitochondrial trafficking in dendrites, and specifically test the hypothesis that PINK1 acts through PKA to increase mitochondrial content by phosphorylating the mitochondrial trafficking adaptor protein Miro2. Aim 3 will determine the mechanisms by which PINK1/PKA activation modulates neurite outgrowth. This work is expected to have an impact on health and human diseases in three areas. First, characterizing the PINK1-PKA signaling pathway will identify new protective mechanisms by which this novel signaling axis maintains dendrite homeostasis. Second, experiments proposed in Aims 1 and 2 will help us understand how PINK1 activates PKA signaling in mitochondria and dendrites regulate mitochondrial trafficking and protect dendrites from oxidative stress. Third, since dysregulation of PKA signaling, mitochondrial function, neurotrophic signaling, and loss of dendrites are implicated in multiple neurodegenerative diseases, identifying new dendrite-protective mechanisms can lead to new targeted, rational therapies via enhanced protective PKA signaling.

项目摘要 PTEN诱导的激酶1（PINK 1）突变与帕金森病的常染色体隐性遗传形式相关 疾病（PD）。在PINK 1基因中，全长PINK 1被蛋白水解加工成低分子量 输出到胞质溶胶的形式。虽然全长PINK 1与调节线粒体自噬有关， 和线粒体功能，了解内源性切割的PINK 1（c- PINK 1）在大脑中的作用是有限的。我们最近的研究表明，c-PINK 1和调节的联系， 线粒体运输和树突生长。该模型得到初步和 我们小组发表的数据表明，体内和体外PINK 1功能的丧失损害了PKA介导的 树突连接和线粒体运输，但这两种机制之间的联系， 悬而未决这些研究将填补我们对c-PINK 1如何调节PKA的理解中的一个关键空白 信号传导以增强PD模型中的树突连接和线粒体运输。在目标1中，分子 PINK 1和树突定位的PKA保护树突免受氧化应激的机制将是 使用基于图像和分子生物学方法阐明。目标2将阐明机制， PINK 1和PKA调节树突中的线粒体运输，并特别测试了以下假设： PINK 1通过PKA通过磷酸化线粒体运输来增加线粒体含量 衔接蛋白Miro 2。目的3将确定PINK 1/PKA激活调节 神经突生长这项工作预计将在三个领域对健康和人类疾病产生影响。第一、 表征PINK 1-PKA信号通路将确定新的保护机制， 信号传导轴维持树突稳态。第二，目标1和2中提出的实验将帮助我们 了解PINK 1如何激活线粒体和树突中的PKA信号传导， 运输和保护树突免受氧化应激。第三，由于PKA信号转导失调， 线粒体功能、神经营养信号传导和树突的丧失与多种神经营养因子有关。 神经退行性疾病，确定新的树突保护机制可以导致新的有针对性的，合理的 通过增强的保护性PKA信号传导治疗。