Transgenic Mice, Inflammation & the Alzheimer Phenotype

转基因小鼠，炎症

基本信息

批准号：
8060487
负责人：
MARCIA N GORDON
金额：
$ 28.68万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-06-01 至 2013-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8060487
关键词：
Adverse reactions Alzheimer&apos s Disease Amyloid Amyloid deposition Area Blood Blood Cells Blood Circulation Bone Marrow Bone Marrow Stem Cell Brain Cell Count Cell Separation Cells Central Nervous System Diseases Chimera organism DNA Data Dependovirus Deposition Development Disease Progression Dose Effectiveness Engineering Enzyme Gene Enzymes Event Gene Delivery Gene Transduction Agent Genome Goals Green Fluorescent Proteins Growth Factor Gene Harvest Home environment Homing Human ITGAM gene Immune Immunotherapy Infiltration Inflammation Inflammatory Injection of therapeutic agent Intravenous Kinetics Knowledge Lead Longevity Magnetism Marrow Methods Minor Mitotic Molecular Mus Neprilysin Nerve Growth Factors Neuraxis Neurodegenerative Disorders Neurosurgical Procedures Operative Surgical Procedures Outcome Pathology Patients Pattern Peptide Hydrolases Phenotype Pilot Projects Population Procedures Process Property Proteins Reaction Relative (related person)Research Risk Role Senile Plaques Site Source Specific qualifier value Spleen Stimulus Testing Therapeutic Tissues Transfection Transgenic Mice Viral Vector Withdrawal base cell type design gene therapy inhibitor/antagonist macrophage monocyte mouse model nerve injury relating to nervous system research study response small molecule success therapeutic gene trafficking tumorigenesis vector

项目摘要

DESCRIPTION (provided by applicant): Alzheimer's disease (AD) has, thus far, resisted therapeutic approaches designed to slow disease progression. Reducing brain amyloid and its consequences is a therapeutic strategy in active development. One proposed means of limiting the damage caused by amyloid is to increase the rate of amyloid clearance using gene therapy. Neprilysin is a major amyloid degrading protease, and a prime candidate for use in anti-amyloid gene therapy. At present, the introduction of therapeutic genes to the brain has been limited to neurosurgical procedures which inject viral vectors directly into the brain. In addition to surgical risks, only small portions of the brain are impacted by these methods. Although many CNS disorders will likely benefit from this approach, the broad distribution of pathology in AD makes delivery of therapeutics by intraparenchymal injections challenging. As an alternative delivery method, we propose to evaluate the use of monocytes, cells normally found in blood, as vectors to transport therapeutic genes to the brain. There are several advantages of monocyte gene therapy. Neurodegenerative disorders cause neural injury and localized inflammation. Circulating monocytes naturally home to these sites of inflammation and concentrate in these areas. This is precisely where the therapeutic gene can be most effective. The therapy is reversible in the event an adverse reaction ensues due to the limited lifespan of the transfected monocytes. The patient's own cells may be harvested for use, reducing the possibility of immune reactions. The monocytes are generally post-mitotic and the transfection method does not integrate new DNA into the genome, reducing the risk of oncogenesis. The procedures involved would be relatively minor, involving intravenous blood withdrawal and reinfusion. In this application we intend to prove the feasibility of using monocyte gene therapy in a transgenic mouse model of amyloid deposition. We will exploit our prior efforts to understand the role of brain macrophages in clearing amyloid plaques, and our preliminary data showing the benefits of neprilysin gene therapy using transfected monocytes. We will identify the best monocyte fraction to use for monocyte gene therapy of the CNS. We will determine the kinetics of monocyte trafficking into the CNS of amyloid depositing mice to develop optimal dosing strategies. We will specify which of several possible mechanisms are active in clearing the amyloid deposits using a genetically engineered, secreted form of neprilysin. Finally, we will test several other amyloid degrading proteases to evaluate whether they may be useful candidates for monocyte gene therapy for AD. Success in the mouse model may rapidly lead to tests of similar gene therapy approaches in AD patients. Alzheimer's disease (AD) has, thus far, resisted therapeutic approaches designed to slow disease progression. Reducing brain amyloid and its consequences is a therapeutic strategy in active development. One proposed means of limiting the damage caused by amyloid is to increase the rate of amyloid clearance using gene therapy. In this application we intend to prove the feasibility of using monocytes, cells which naturally home to sites of inflammation, as carriers for gene therapy in a transgenic mouse model of amyloid deposition.

描述（由申请人提供）：迄今为止，阿尔茨海默病（AD）一直对旨在减缓疾病进展的治疗方法产生抵制。减少大脑淀粉样蛋白及其后果是积极发展中的一种治疗策略。一种限制淀粉样蛋白造成的损害的建议方法是使用基因疗法提高淀粉样蛋白的清除率。脑啡肽酶是一种主要的淀粉样蛋白降解蛋白酶，也是用于抗淀粉样蛋白基因治疗的主要候选酶。目前，将治疗基因引入大脑仅限于将病毒载体直接注入大脑的神经外科手术。除了手术风险之外，这些方法仅影响大脑的一小部分。尽管许多中枢神经系统疾病可能会受益于这种方法，但 AD 病理学的广泛分布使得通过实质内注射进行治疗具有挑战性。作为一种替代的递送方法，我们建议评估单核细胞（通常存在于血液中的细胞）作为将治疗基因转运到大脑的载体的使用。单核细胞基因治疗有几个优点。神经退行性疾病会导致神经损伤和局部炎症。循环单核细胞自然地驻留在这些炎症部位并集中在这些区域。这正是治疗基因最有效的地方。如果由于转染的单核细胞的寿命有限而发生不良反应，则该疗法是可逆的。可以采集患者自身的细胞进行使用，从而减少免疫反应的可能性。单核细胞通常是有丝分裂后的，转染方法不会将新的DNA整合到基因组中，从而降低了肿瘤发生的风险。所涉及的程序相对较小，包括静脉抽血和回输。在本申请中，我们打算证明在淀粉样蛋白沉积的转基因小鼠模型中使用单核细胞基因治疗的可行性。我们将利用我们之前的努力来了解脑巨噬细胞在清除淀粉样斑块中的作用，以及我们的初步数据显示使用转染的单核细胞进行脑啡肽酶基因治疗的益处。我们将确定用于中枢神经系统单核细胞基因治疗的最佳单核细胞分数。我们将确定单核细胞转运至淀粉样蛋白沉积小鼠中枢神经系统的动力学，以制定最佳剂量策略。我们将具体说明几种可能的机制中哪一种在使用基因工程、分泌形式的脑啡肽酶清除淀粉样蛋白沉积物方面发挥着积极作用。最后，我们将测试其他几种淀粉样蛋白降解蛋白酶，以评估它们是否可能成为 AD 单核细胞基因治疗的有用候选者。小鼠模型的成功可能会迅速导致在 AD 患者中进行类似基因治疗方法的测试。迄今为止，阿尔茨海默病（AD）一直对旨在减缓疾病进展的治疗方法产生抵制。减少大脑淀粉样蛋白及其后果是积极发展中的一种治疗策略。一种限制淀粉样蛋白造成的损害的建议方法是使用基因疗法提高淀粉样蛋白的清除率。在此应用中，我们打算证明使用单核细胞（自然定位于炎症部位的细胞）作为淀粉样蛋白沉积转基因小鼠模型中基因治疗的载体的可行性。