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）是一种常规用于血液学/肿瘤学环境的多模式造血生长因子，但文献中出现了有关其对脑部疾病有益的报道。 在我们实验室对阿尔茨海默病（AD）小鼠模型的初步研究表明，它具有促认知和疾病修饰作用。 负责这些深刻的影响的机制尚不清楚，G-CSF作为AD患者的治疗剂的开发需要临床前研究。 目的：拟定的研究旨在阐明G-CSF降低AD小鼠模型中淀粉样蛋白沉积/水平、刺激海马神经发生和恢复突触功能活性的细胞和分子机制。 目标1：将在骨髓来源的细胞表达绿色荧光蛋白（GFP）的嵌合tg AD小鼠中测试骨髓来源的细胞从血液向脑的运输增强和/或小胶质细胞和吞噬细胞活性增加是β-淀粉样蛋白（A-2）沉积减少的原因的假设。 目标二：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这一假设，将用神经解剖学和电生理学技术测试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，机制研究将阐明由G-CSF触发的新的信号传导途径，其涉及转换为蛋白激酶C（PKC-δ 2）的可变剪接变体，其刺激海马神经发生并抑制Tau的多磷酸化，这两者被假定为改善海马突触功能的基础。 研究方法：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。 将产生植入表达绿色荧光蛋白（GFP+）的骨髓细胞的嵌合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。 迄今为止的发现：给予认知受损的tg AD小鼠（APP+PS1）G-CSF逆转了 认知能力受损，海马和内嗅皮层A2沉积显著减少。 G-CSF治疗的tg AD小鼠也显示了总的小胶质细胞增生和海马CA 1和CA 3区突触素免疫染色面积的增加显着增加。 从给予tg AD小鼠G-CSF获得的其他初步结果包括海马神经发生的刺激和海马切片电生理功能的改善。 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和刺激健康骨髓供体的造血干细胞生成。 G-CSF在AD小鼠模型中的认知增强和潜在的疾病修饰作用为临床试验提供了强大的动力，以逆转或阻止AD患者痴呆的进展。 与VA使命的相关性：AD是一种无法治愈的年龄依赖性痴呆疾病，在美国的患病率为500万。 随着退伍军人人口的老龄化，AD的患病率将增加，导致对VA医疗保健系统的压倒性需求。 从人道主义和社会经济的角度来看，迫切需要改善疾病的药物来延迟或逆转AD的进行性痴呆。 公共卫生相关性： Filgastrim（G-CSF）是一种多模式造血生长因子，对中枢神经系统疾病也有深远的影响。 我们实验室最近的工作表明，给予认知受损的阿尔茨海默病（AD）转基因（tg）小鼠G-CSF可逆转认知能力，并显著减少海马和内嗅皮质中的淀粉样蛋白（A <$）沉积。 G-CSF治疗的tg AD小鼠也显示了总的小胶质细胞增生和突触囊泡蛋白免疫反应性在海马CA 1和CA 3区的面积增加显着增加。 本研究项目的总体目标是阐明G-CSF在Tg APP/PS1小鼠中的疾病修饰作用。 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产生，该研究项目的结果将有助于将G-CSF转化为临床试验，以逆转或阻止AD中痴呆的进展。