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: 8397542
• 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 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8397542
• 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; abstracting; 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; receptor; relating to nervous system; restoration; socioeconomics; stem; synaptic function; tau Proteins; tau phosphorylation; trafficking

Abstract/Summary 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-¿) 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 hippocamapal 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 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 immunostaining in hippocamal 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.

摘要/总结 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 常规用于血液学/肿瘤学环境，但有关其对脑部疾病有益的报道是 出现在文献中。 我们在阿尔茨海默病（AD）小鼠模型中进行的初步研究 实验室表明它具有促进认知和缓解疾病的作用。 机制 这些深远影响的原因尚不清楚，G-CSF 作为治疗剂的开发 对于 AD 患者来说，需要进行临床前研究。 目的：拟议的研究旨在通过以下方式阐明细胞和分子机制： 其中 G-CSF 减少淀粉样蛋白沉积/水平，刺激海马神经发生并恢复 AD 小鼠模型中的突触功能活动。 目标 1：加强贩运的假设 从血液到大脑的骨髓衍生细胞和/或小胶质细胞和吞噬细胞活性的增加 将在嵌合 tg AD 小鼠中测试导致 β-淀粉样蛋白 (A-¿) 沉积减少的原因 其中骨髓来源的细胞表达绿色荧光蛋白（GFP）。 目标 2：测试 假设 G-CSF 的促认知作用是通过对神经祖细胞的直接作用介导的 细胞和神经元，G-CSF 促进神经发生和/或促进恢复的程度 海马突触功能和结构将通过神经解剖学和 电生理学技术。 目标 3：在细胞内水平上，机制研究将阐明 由 G-CSF 触发的新信号通路，涉及切换到选择性剪接变体 蛋白激酶 C (PKC-delta2) 刺激海马神经发生并抑制多聚 Tau 磷酸化，这两者被认为是海马突触改善的基础 功能。 方法：细胞和分子生物学技术、免疫组织化学、海马 将采用切片电生理学和行为分析。 嵌合 tg AD 小鼠植入 将产生表达绿色荧光蛋白（GFP+）的骨髓细胞以确定 外周来源的小胶质细胞和其他骨髓来源的细胞在多大程度上有助于 淀粉样蛋白减少和/或神经发生。 迄今为止的发现：对认知受损的 tg AD 小鼠 (APP+PS1) 施用 G-CSF 可以逆转 认知能力受损并显着减少海马和 内嗅皮层。 G-CSF 治疗的 tg AD 小鼠还显示，总小胶质细胞增生和 海马 CA1 和 CA3 区域突触素免疫染色面积增加。 额外的 对 tg AD 小鼠施用 G-CSF 获得的初步结果包括刺激 海马神经发生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并刺激 健康骨髓捐赠者的造血干细胞生成。 认知增强和 G-CSF 在 AD 小鼠模型中的潜在疾病缓解作用为 逆转或预防 AD 患者痴呆进展的临床试验。 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，患病率为 5 万在美国。 随着退伍军人人口老龄化，AD 患病率将会增加，导致 对 VA 医疗保健系统的巨大需求。 从人道主义和社会经济两方面 从观点来看，迫切需要能够延迟或逆转疾病的疾病缓解药物 AD 进行性痴呆。