Investigating the role of neuronal SYNJ2 in mRNA transport and mitochondrial function

研究神经元 SYNJ2 在 mRNA 转运和线粒体功能中的作用

• 批准号: 10747226
• 负责人: Whitney Sharee Gibbs
• 金额: $ 5.2万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10747226
• 关键词: Advisory Committees; Alzheimer' s Disease; Axon; Back; Cell Differentiation process; Cells; Complex; Dendrites; Disease; Distal; Distant; Ensure; Goals; Half-Life; Health; Heterogeneity; Homeostasis; Human; Huntington Disease; Laboratories; Maintenance; Mediating; Membrane; Mentors; Messenger RNA; Mitochondria; Mitochondrial Proteins; Modeling; Mus; Mutate; Nerve Degeneration; Neurons; PINK1 gene; Parkinson Disease; Pathologic; Peripheral; Phase; Phosphotransferases; Physiological; Play; Process; Protein Biosynthesis; Proteins; RNA Binding; RNA Recognition Motif; Research; Role; SYNJ1 gene; Surface; Training; Transcript; Translations; Travel; Work; axon regeneration; axonal degeneration; career; diversity and inclusion; in vivo regeneration; innovation; meter; mitochondrial messenger RNA; neuronal cell body; neurotransmission; preservation

Project Summary Neurons critically depend on mitochondria function to maintain membrane excitability and execute complex functions, such as neurotransmission and plasticity. Neurons are highly differentiated cells that require large amounts of ATP to perform these functions and ensure long-term viability. The unique complexity of neurons is reflected by the extremely long segments that can extend up to a meter long and the functional heterogeneity for each neuronal compartment. The neuron has specialized mechanisms to transport mitochondria to the most distal parts to maintain proper neuronal function and survival. In turn, peripheral mitochondria rely on the transport of cellular components, such as mRNA and proteins, to sustain mitochondrial homeostasis without the need to travel back to the soma. The current proposal focuses on this aspect of mitochondrial maintenance, mitochondrial transcripts' ability to be trafficked to the axons for local translation, and how this influences mitochondrial and neuronal health and function. This project's significance is focused on PINK1, an essential mitochondrial kinase that is mutated in a hereditable form of Parkinson's disease. PINK1 protein will not survive transport down the axon because of its short half-life. To this end, our laboratory described an innovative mechanism by which mitochondria carry PINK1 mRNA on its surface to axons. Synaptojanin 2 (SYNJ2) was found to be responsible for tethering PINK1 mRNA to the mitochondria for axonal localization and local protein synthesis. The study of this neuronal-specific model suggests that the RNA binding function of SYNJ2 is required for PINK1-mediated processes (such as mitophagy); however, this has yet to be explored. The work planned in this proposal will explore the physiological and pathological consequence of disrupting the RNA binding function of SYNJ2, and; this unique approach is of critical importance for understanding mitochondrial mRNA transport and translation for preserving mitochondrial health. Thus, I hypothesize that Hypothesis: The RNA Recognition Motif of SYNJ2 causes PINK1 mRNA to colocalize with and be transported with mitochondria in a manner critical for PINK1 functions in axons and dendrites and thereby for maintaining neuronal health. I have assembled an advisory committee to provide conceptual and technical guidance as I explore the following Specific Aims: Aim 1: Examine the function of endogenous SYNJ2 and its RNA recognition motif in mice. Aim 2: Establish the role of peripheral SYNJ2 in mediating axonal mitochondrial function and neuronal health Aim 3: Investigate the impact of SYNJ2 RNA binding function in modulating axonal degeneration and regeneration in vivo. I have also developed a tailored diversity and inclusion training plan to execute during the mentored phase. The proposed studies and mentoring plans described in this proposal will provide me with a robust training platform to launch my independent academic research career.

项目摘要 神经元严重依赖线粒体功能来维持膜兴奋性并执行复合物 功能，例如神经传递和可塑性。神经元是高度分化的细胞，需要大的细胞 执行这些功能并确保长期生存能力的ATP量。神经元的独特复杂性是 由极长的段反映出可以延伸至米长的且功能异质性 对于每个神经元室。神经元具有将线粒体运输到的专门机制 大多数远端部分保持适当的神经元功能和生存。反过来，外围线粒体依赖于 细胞成分（例如mRNA和蛋白质）的运输，以维持线粒体稳态而没有 需要回到索马。当前的建议重点是线粒体的这一方面 维护，线粒体笔录的能力被贩运到轴突进行本地翻译，以及如何 影响线粒体和神经元健康和功能。 该项目的意义集中在Pink1上，PINK1是一种必需的线粒体激酶，在A中突变 帕金森氏病的可遗传形式。 Pink1蛋白无法生存在轴突下运输的 半衰期。为此，我们的实验室描述了线粒体携带的创新机制 粉红色的mRNA表面与轴突。发现Synaptojanin 2（Synj2）是束缚的原因 PINK1 mRNA到线粒体的轴突定位和局部蛋白质合成。这项研究 神经元特异性模型表明，Synj2的RNA结合函数是pink1介导的 过程（例如线粒体）；但是，这尚未探索。该提案中计划的工作将 探索破坏synj2的RNA结合功能的生理和病理后果， 和;这种独特的方法对于理解线粒体mRNA运输至关重要 保留线粒体健康的翻译。 因此，我假设该假设：Synj2的RNA识别基序使Pink1 mRNA达到 以对轴突中pink1函数至关重要的方式与线粒体共定位并与线粒体一起运输 和树突，从而维持神经元健康。我已经组建了一个咨询委员会 在我探索以下特定目的时，提供概念和技术指导：目标1：检查 内源Synj2及其RNA识别基序的功能。目标2：确定外围的作用 Synj2中介导轴突线粒体功能和神经元健康目标3：研究Synj2的影响 RNA结合函数在调节体内的轴突变性和再生中。我也开发了 量身定制的多样性和包容培训计划在指导阶段执行。拟议的研究和 该提案中描述的指导计划将为我提供一个强大的培训平台，以启动我的 独立的学术研究职业。