Disease-Modifying Effects of Filgastrim in a Mouse Model of AD

非加司亭对 AD 小鼠模型的疾病缓解作用

• 批准号: 7797233
• 负责人: JUAN R SANCHEZ-RAMOS
• 金额: --
• 依托单位: JAMES A. HALEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-10-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7797233
• 关键词: APP-PS1; Accounting; Aftercare; Age; Alternative Splicing; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-Protein; Amyloid deposition; Apoptosis; Area; Behavioral; Birth; Blood Cells; Bone Marrow; Bone Marrow Cells; Bone Marrow Stem Cell; Brain; Brain Diseases; Bromodeoxyuridine; CSF3 gene; Cells; Cessation of life; Clinical Trials; Cognition Disorders; Cognitive; Dementia; Dendritic Spines; Deposition; Development; Disease; Electrophysiology (science); Event; Generations; Golgi Apparatus; Granulocyte Colony-Stimulating Factor; Green Fluorescent Proteins; Healthcare Systems; Hematology; Hematopoietic Cell Growth Factors; Hematopoietic stem cells; Hippocampus (Brain); Immunohistochemistry; Impaired cognition; In Vitro; Infiltration; Laboratories; Learning; Literature; Long-Term Depression; Long-Term Potentiation; Measures; Mediating; Methods; Microglia; Mission; Molecular; Molecular Biology Techniques; Morphology; Mus; Neuraxis; Neurodegenerative Disorders; Neurons; Neutropenia; Outcome; Pathogenesis; Patients; Performance; Pharmaceutical Preparations; Phosphorylation; Physiological; Population; Prevalence; Production; Protein Isoforms; Protein Kinase C; RNA Splicing; Recovery; Reporting; Research Design; Research Project Grants; Schedule; Signal Pathway; Signal Transduction; Slice; Staining method; Stains; Stem cells; Structure; Synapses; Synaptophysin; Techniques; Testing; Therapeutic; Therapeutic Agents; Transgenic Mice; Transgenic Organisms; Translations; Variant; Veterans; Work; age related; brain repair; clinically relevant; cytokine; design; entorhinal cortex; granulocyte; immunoreactivity; improved; in vivo; light treatment; macrophage; mimetics; mouse model; nerve stem cell; neurofilament; neurogenesis; new therapeutic target; novel; oncology; pre-clinical research; progenitor; programs; public health relevance; receptor; relating to nervous system; restoration; socioeconomics; stem; synaptic function; tau Proteins; tau phosphorylation; trafficking

DESCRIPTION (provided by applicant): Filgastrim (granulocyte colony stimulating factor or G-CSF) is a multi-modal hematopoietic growth factor used routinely in the hematology/oncology setting but reports of its benefits for brain diseases are appearing in the literature. Preliminary studies in a mouse model of Alzheimer's disease (AD) in our laboratory suggests it has both pro-cognitive and disease-modifying effects. The mechanisms responsible for these profound effects are not clear and development of G-CSF as a therapeutic agent for patients with AD warrants pre-clinical research. Objectives: The proposed studies are designed to elucidate the cellular and molecular mechanisms by which G-CSF reduces amyloid deposition/levels, stimulates hippocampal neurogenesis and restores synaptic functional activity in a mouse model of AD. Aim 1: The hypothesis that enhanced trafficking of bone-marrow derived cells from blood to brain and/or increased microglial and phagocytic activity is responsible for the decrease in beta-amyloid (A-2) deposition will be tested in a chimeric tg AD mouse in which bone marrow derived cells express green fluorescent protein (GFP). Aim 2: To test the hypothesis that the pro-cognitive effects of G-CSF are mediated by direct actions on neural progenitor cells and neurons, the extent to which G-CSF promotes neurogenesis and/or promotes recovery of hippocampal synaptic function and structure will be tested with neuroanatomical and electrophysiological techniques. Aim 3: At an intracellular level, mechanistic studies will elucidate a novel signaling pathway triggered by G-CSF that involves switching to an alternatively spliced variant of protein kinase C (PKC-delta2) that stimulates hippocampal neurogenesis and inhibits poly- phosphorylation of Tau, both which are postulated to underlie improved hippocampal synaptic function. Methods: Cellular and molecular biology techniques, immunohistochemistry, hippocampal slice electrophysiology and behavioral analysis will be employed. Chimeric tg AD mice engrafted with green fluorescent protein expressing (GFP+) bone marrow cells will be generated to determine the extent to which peripherally derived microglia and other bone marrow-derived cells contribute to amyloid reduction and/or neurogenesis. Findings to date: Administration of G-CSF to cognitively-impaired tg AD mice (APP+PS1) reversed the impaired cognitive performance and significantly decreased A2 deposition in hippocampus and entorhinal cortex. G-CSF-treated tg AD mice also revealed a significant increase in total microgliosis and increased areas of synaptophysin immunostaining in hippocampal CA1 and CA3 regions. Additional preliminary results obtained from G-CSF administration to tg AD mice include stimulation of hippocampal neurogenesis and improvement of electrophysiological function in hippocampal slices. Clinical relevance: G-CSF is routinely and safely used to treat neutropenia and to stimulate hematopoietic stem cell generation in healthy bone marrow donors. The cognitive-enhancing and potential disease-modifying effects of G-CSF in a mouse model of AD provides a strong impetus for clinical trials to reverse or forestall progression of dementia in AD patients. Relevance to VA Mission: AD is an incurable, age-dependent dementing disease with a prevalence of 5 million in the US. As the veteran population ages, the prevalence of AD will increase, resulting in overwhelming demands on the VA health care system. From both humanitarian and socio-economic perspectives, there is an urgent need for disease-modifying medications that delay or reverse the progressive dementia of AD. PUBLIC HEALTH RELEVANCE: Filgastrim (G-CSF) is a multi-modal hematopoietic growth factor, which also has profound effects on the diseased central nervous system. Recent work in our laboratory has demonstrated that administration of G-CSF to cognitively-impaired Alzheimer's disease (AD) transgenic (tg) mice reversed cognitive performance and significantly decreased ¿-amyloid (A¿) deposition in hippocampus and entorhinal cortex. G-CSF-treated tg AD mice also revealed a significant increase in total microgliosis and increased areas of synaptophysin immunoreactivity in hippocamal CA1 and CA3 regions. The overall objective of this research project is to elucidate the disease-modifying effects of G-CSF in Tg APP/PS1 mice. We will study events triggered and mediated by G-CSF at the electrophysiological, cellular, and molecular levels that contribute to amyloid reduction, restoration of synaptic activity, hippocampal neurogenesis and cognitive improvement. Given that G-CSF is already utilized clinically to stimulate hematopoietic stem cell production, results from this research project will facilitate translation into clinical trials of G-CSF to reverse or forestall the progression of dementia in AD.

描述（由申请人提供）： Filgastrim（粒细胞集落刺激因子或 G-CSF）是一种多模式造血生长因子，常用于血液学/肿瘤学领域，但文献中也出现了其对脑部疾病有益的报道。 我们实验室对阿尔茨海默病 (AD) 小鼠模型的初步研究表明，它具有促进认知和缓解疾病的作用。 造成这些深远影响的机制尚不清楚，G-CSF 作为 AD 患者治疗剂的开发值得临床前研究。 目的：拟议的研究旨在阐明 G-CSF 减少 AD 小鼠模型中淀粉样蛋白沉积/水平、刺激海马神经发生并恢复突触功能活动的细胞和分子机制。 目标 1：骨髓来源细胞从血液到大脑的运输增强和/或小胶质细胞和吞噬细胞活性增加是导致 β-淀粉样蛋白 (A-2) 沉积减少的原因，这一假设将在嵌合 tg AD 小鼠中进行测试，其中骨髓来源细胞表达绿色荧光蛋白 (GFP)。 目标 2：为了检验 G-CSF 的促认知作用是通过对神经祖细胞和神经元的直接作用介导的假设，将使用神经解剖学和电生理学技术来测试 G-CSF 促进神经发生和/或促进海马突触功能和结构恢复的程度。 目标 3：在细胞内水平上，机制研究将阐明由 G-CSF 触发的一种新的信号传导途径，该途径涉及转换为蛋白激酶 C (PKC-delta2) 的选择性剪接变体，刺激海马神经发生并抑制 Tau 多磷酸化，这两者被认为是改善海马突触功能的基础。 方法：将采用细胞和分子生物学技术、免疫组织化学、海马切片电生理学和行为分析。 将产生植入绿色荧光蛋白表达（GFP+）骨髓细胞的嵌合tg AD小鼠，以确定外周来源的小胶质细胞和其他骨髓来源的细胞对淀粉样蛋白减少和/或神经发生的贡献程度。 迄今为止的研究结果：对认知受损的 tg AD 小鼠 (APP+PS1) 施用 G-CSF 可逆转受损的认知能力，并显着减少海马和内嗅皮质中的 A2 沉积。 G-CSF 治疗的 tg AD 小鼠还显示出海马 CA1 和 CA3 区域小神经胶质细胞总数显着增加，突触素免疫染色面积增加。 对 tg AD 小鼠施用 G-CSF 获得的其他初步结果包括刺激海马神经发生和改善海马切片的电生理功能。 临床相关性：G-CSF 常规且安全地用于治疗中性粒细胞减少症并刺激健康骨髓捐赠者的造血干细胞生成。 G-CSF 在 AD 小鼠模型中的认知增强和潜在的疾病缓解作用为逆转或预防 AD 患者痴呆进展的临床试验提供了强大的推动力。 与 VA 使命的相关性：AD 是一种无法治愈的、与年龄相关的痴呆症，在美国有 500 万人患病。 随着退伍军人人口老龄化，阿尔茨海默病的患病率将会增加，导致对退伍军人管理局医疗保健系统的巨大需求。 从人道主义和社会经济角度来看，迫切需要能够延缓或逆转 AD 进行性痴呆的疾病缓解药物。 公共卫生相关性： Filgastrim (G-CSF) 是一种多模式造血生长因子，对患病的中枢神经系统也有深远的影响。 我们实验室最近的工作表明，对认知受损的阿尔茨海默病 (AD) 转基因 (tg) 小鼠施用 G-CSF 可逆转认知能力，并显着减少海马和内嗅皮层中的 β-淀粉样蛋白 (A¿) 沉积。 G-CSF 治疗的 tg AD 小鼠还显示，海马 CA1 和 CA3 区域的小胶质细胞总数显着增加，突触素免疫反应区域也增加。 该研究项目的总体目标是阐明 G-CSF 对 Tg APP/PS1 小鼠的疾病缓解作用。 我们将在电生理、细胞和分子水平上研究 G-CSF 触发和介导的事件，这些事件有助于淀粉样蛋白减少、突触活性恢复、海马神经发生和认知改善。 鉴于 G-CSF 已在临床上用于刺激造血干细胞的产生，该研究项目的结果将有助于转化为 G-CSF 的临床试验，以逆转或预防 AD 痴呆的进展。