Investigating the Role of GDE2 in Neuronal Mitotic Inhibition and Nuclear Pore Complex Defects

研究 GDE2 在神经元有丝分裂抑制和核孔复合体缺陷中的作用

• 批准号: 10231661
• 负责人: Anna Nicole Westerhaus
• 金额: $ 4.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-13 至 2024-04-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10231661
• 关键词: ALS patients; Address; Adult; Affect; Aging; Alzheimer' s Disease; Alzheimer' s disease brain; Amyotrophic Lateral Sclerosis; Autopsy; Brain; Cell Cycle; Cell Cycle Inhibition; Cell Cycle Progression; Cell Cycle Regulation; Cell Proliferation; Cell surface; Cellular biology; Chronology; Cleaved cell; Data; Defect; Dementia; Development; Disease; Dissection; Embryo; Embryonic Development; Enzymes; Etiology; Exhibits; Functional disorder; GPI Membrane Anchors; Genetic Epistasis; Grant; Human; Immunohistochemistry; Impairment; In Vitro; Knock-out; Knockout Mice; Knowledge; Life; Maintenance; Mediating; Membrane; Mentors; Mentorship; Mitosis; Mitotic; Mitotic Cell Cycle; Molecular; Motor Cortex; Mus; National Research Service Awards; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Neurosciences; Nuclear Envelope; Nuclear Pore Complex; Outcome; Pathogenesis; Pathologic; Pathology; Pathway interactions; Phenotype; Physiological; Primary Cell Cultures; Process; Proteins; Public Health; RNA analysis; Research; Role; Sampling; Scientist; Severities; Signal Transduction; Structure; Technical Expertise; Testing; Therapeutic Intervention; Time; Training; Transmission Electron Microscopy; Ubiquitination; Viral; Virus; WNT Signaling Pathway; Work; base; brain tissue; collaborative environment; design and construction; effective therapy; experimental study; glycerophosphodiester phosphodiesterase; human disease; human tissue; improved; in vivo; insight; knockout animal; mouse genetics; neuronal survival; new therapeutic target; novel; nucleocytoplasmic transport; prevent; skills; stem cell proliferation; teacher; therapeutic candidate; tissue preparation; transcriptome sequencing

Project Summary / Abstract Alzheimer's disease (AD) and related diseases (ADRDs) are the leading cause of dementia, affecting millions of people each year. Despite being such a prevalent public health concern, knowledge of the etiology of neurodegeneration in these diseases is limited and has stymied the development of effective treatments. Aberrant neuronal cell-cycle reentry (CCR) and nuclear pore complex (NPC)/nucleocytoplasmic transport defects have been separately identified as causal phenomena for neurodegeneration in AD and ADRDs. However, the mechanisms leading to these phenomena and the relationship between them remains unknown. We have identified a pathway involving Glycerophosphodiester phosphodiesterase 2 (GDE2 or GDPD5) that prevents CCR and NPC/nucleocytoplasmic transport defects in the adult mammalian brain and is disrupted in human postmortem AD and ALS brain tissue. GDE2 is one of three six-transmembrane enzymes that act at the cell surface to cleave the GPI-anchor that tethers some proteins to the membrane. During embryogenesis, GDE2 inhibits progenitor cell proliferation to promote neuronal differentiation. Preliminary studies show that loss of GDE2 in the adult brain leads to CCR and NPC abnormalities that precede overt neurodegeneration. Because NPC defects are coincident with CCR in neurons lacking GDE2, and the NPC normally breaks down during mitosis, we hypothesize that GDE2 is important for maintaining neurons in a postmitotic state throughout life and that aberrant CCR elicited by GDE2 loss leads to NPC breakdown and neurodegeneration. Importantly, GDE2 localization and function is disrupted in AD and ADRDs, raising the possibility that dysfunction of the GDE2 pathway could contribute to the pathogenesis of these diseases. This proposal will test our central hypothesis in three aims. Aim 1 will define the physiological requirement for GDE2 in maintaining neuronal quiescence and NPC integrity in aging mice while Aim 2 will define the relationship between CCR and NPC breakdown and identify the mechanisms leading to CCR and NPC defects. Later experiments will determine relevance of this pathway to CCR and NPC disruption in human AD and ADRDs (Aim 3). The proposed research is expected to identify a novel physiological pathway that is essential for neuronal survival and will provide new insight into causal mechanisms of neurodegeneration that are relevant to human disease. In addition, this project has immense training potential. I will have the opportunity to improve my knowledge in neuroscience and cell biology, and to acquire technical expertise in primary cell culture, viral construct design and transduction, mouse genetics, brain dissections and tissue preparation, and immunohistochemical analysis of human tissues. This research will be performed in a highly collaborative environment, where I will have numerous opportunities to receive quality mentorship and training, to develop my written and presentation skills, and to grow as a mentor and teacher to more junior scientists. Overall, the Kirschstein-NRSA grant will support both my research aimed at discovering the molecular basis of neurodegeneration and my development as an independent scientist.

项目摘要 /摘要 阿尔茨海默氏病（AD）和相关疾病（ADRD）是痴呆症的主要原因，影响了数百万 每年的人。尽管公共卫生如此普遍，但对 这些疾病中的神经变性受到限制，并阻碍了有效治疗的发展。 异常的神经元细胞周期再入（CCR）和核孔复合物（NPC）/核总质转运 缺陷已被分别识别为AD和ADRD中神经变性的因果现象。 但是，导致这些现象及其之间的关系的机制仍然未知。 我们已经确定了涉及甘油磷酸二酯磷酸二酯酶2（GDE2或GDPD5）的途径 防止成人哺乳动物大脑中的CCR和NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/NPC/ 人验尸广告和ALS脑组织。 GDE2是在对 细胞表面裂解将一些蛋白质转移到膜上的GPI锚定。在胚胎发生期间，GDE2 抑制祖细胞增殖以促进神经元分化。初步研究表明失去 成年大脑中的GDE2导致CCR和NPC异常，在明显的神经变性之前。因为 NPC缺陷与缺乏GDE2的神经元中的CCR一致，NPC通常在期间分解 有丝分裂，我们假设GDE2对于在一生中保持神经元的神经元很重要，并且 GDE2损耗引起的异常CCR导致NPC崩溃和神经变性。重要的是，GDE2 AD和ADRD中的本地化和功能受到破坏，从而提高了GDE2功能障碍的可能性 途径可能有助于这些疾病的发病机理。该提议将检验我们在 三个目标。 AIM 1将定义GDE2的生理要求，以保持神经元静止和 衰老小鼠中的NPC完整性，而AIM 2将定义CCR与NPC分解之间的关系 确定导致CCR和NPC缺陷的机制。以后的实验将确定这一点的相关性 人类AD和ADRD中CCR和NPC中断的途径（AIM 3）。拟议的研究有望 确定一种对神经元生存至关重要的新型生理途径，并将为您提供新的见解 与人类疾病有关的神经变性的因果机制。此外，这个项目有 巨大的训练潜力。我将有机会提高我在神经科学和细胞生物学方面的知识， 并获得原代细胞培养，病毒构建设计和转导的技术专长，小鼠遗传学， 脑解剖和组织制备以及人体组织的免疫组织化学分析。这项研究 将在高度协作的环境中进行，我将有很多机会接收 质量指导和培训，发展我的书面和演讲技巧，并成长为导师和 教师的更多初级科学家。总体而言，Kirschstein-Nrsa Grant将支持我的研究针对 发现神经退行性的分子基础和我作为独立科学家的发展。