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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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.

项目概要/摘要阿尔茨海默病 (AD) 和相关疾病 (ADRD) 是导致痴呆症的主要原因，影响着数百万人每年的人。尽管这是一个如此普遍的公共卫生问题，但对病因的了解这些疾病中的神经变性是有限的，并且阻碍了有效治疗的开发。异常神经元细胞周期折返 (CCR) 和核孔复合体 (NPC)/核细胞质转运缺陷已被分别确定为 AD 和 ADRD 神经变性的因果现象。然而，导致这些现象的机制以及它们之间的关系仍然未知。我们已经确定了一条涉及甘油磷酸二酯磷酸二酯酶 2（GDE2 或 GDPD5）的途径，该途径防止成年哺乳动物大脑中的 CCR 和 NPC/核细胞质运输缺陷，并在人类死后 AD 和 ALS 脑组织。 GDE2 是三种六跨膜酶之一，作用于细胞表面裂解将某些蛋白质束缚在细胞膜上的 GPI 锚。在胚胎发生过程中，GDE2 抑制祖细胞增殖，促进神经元分化。初步研究表明，损失成人大脑中的 GDE2 会导致 CCR 和 NPC 异常，从而导致明显的神经变性。因为 NPC 缺陷与缺乏 GDE2 的神经元中的 CCR 一致，并且 NPC 通常在有丝分裂中，我们假设 GDE2 对于维持神经元一生处于有丝分裂后状态很重要，并且 GDE2 缺失引起的异常 CCR 会导致 NPC 崩溃和神经退行性变。重要的是，GDE2 AD 和 ADRD 中的定位和功能被破坏，增加了 GDE2 功能障碍的可能性途径可能有助于这些疾病的发病机制。该提案将检验我们的中心假设三个目标。目标 1 将定义 GDE2 在维持神经元静止和衰老小鼠的 NPC 完整性，而目标 2 将定义 CCR 与 NPC 分解和确定导致 CCR 和 NPC 缺陷的机制。稍后的实验将确定这一点的相关性人类 AD 和 ADRD 中 CCR 和 NPC 破坏的途径（目标 3）。拟议的研究预计将确定对神经元生存至关重要的新生理途径，并将提供新的见解与人类疾病相关的神经变性的因果机制。此外，该项目还有巨大的培训潜力。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、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，在那里我将有很多机会接受高质量的指导和培训，培养我的书面和演讲技巧，并成长为一名导师和为更多初级科学家提供指导。总的来说，Kirschstein-NRSA 赠款将支持我的研究目标发现神经退行性变的分子基础以及我作为一名独立科学家的发展。