AAV-mediated muscle-directed gene transfer for Alzheimer's disease

AAV 介导的肌肉定向基因转移治疗阿尔茨海默病

• 批准号: 7433188
• 负责人: Ken-ichiro Fukuchi
• 金额: $ 19.37万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7433188
• 关键词: AN-1792; APP gene; Adverse effects; Age; Alzheimer' s Disease; Alzheimer' s disease model; Amino Acids; Amyloid; Amyloid deposition; Antibodies; Appendix; Attenuated; Behavioral; Biochemical; Blood Circulation; Blood Vessels; Brain; Cerebrum; Clinical Trials; Cognitive deficits; Dementia; Deposition; Elderly; Encephalitis; Epitopes; Etiology; Fc Receptor; Goals; Human; Immune; Immune response; Immunization; Immunotherapy; Inflammatory Response; Injection of therapeutic agent; Knock-out; Learning; Mediating; Memory; Memory impairment; Modality; Monitor; Mus; Muscle; Mutation; Needles; Neurodegenerative Disorders; Neurofibrillary Tangles; Passive Immunization; Pathogenesis; Pathology; Patients; Peptides; Peripheral; Play; Prevention; Production; Protein Overexpression; Recombinant adeno-associated virus (rAAV); Reporting; Role; Safety; Screening procedure; Senile Plaques; Serum; Skeletal Muscle; T-Lymphocyte; Therapeutic Effect; Thigh structure; Topical application; Toxic effect; Transgenic Mice; amyloid peptide; base; cell mediated immune response; familial Alzheimer disease; gene therapy; immunoreactivity; improved; mouse model; mutant; neuron loss; novel; peptide A; presenilin-1; prevent; prophylactic; vector

DESCRIPTION (provided by applicant): Alzheimer's Disease (AD) is the most common cause of dementia after the age of 60. The pathological hallmarks of Alzheimer's disease include deposition of amyloid ¿-peptide (A¿) in neuritic plaques and cerebral blood vessels, neurofibrillary tangles, and loss of neurons. Increasing evidence supports the notion that A¿ and its precursor play important roles in the pathogenesis of Alzheimer's disease. Immunization of mouse models of Alzheimer's disease with synthetic A¿ reduces A¿ deposits and attenuates their memory and learning deficits. Recent clinical trials, however, were halted due to brain inflammation, presumably induced by T-cell mediated and/or Fc-mediated immune responses. Peripheral administration of antibodies against A¿ also induced clearance of preexisting amyloid plaques in AD mouse models, indicating that an active T-cell-mediated immune response is unnecessary. Topical application of the F(ab')2 without Fc of antibodies against A¿ led to clearance of amyloid deposits in an AD mouse model, indicating that non-Fc-mediated mechanisms are also involved in the clearance. We have demonstrated that human single chain antibody (scFv) against A¿ was also effective in reducing brain A¿ load in an AD mouse model. Although these passive immunization modalities may be effective in treating AD patients without inducing side effects such as inflammatory responses, such modalities suffer from repeated administrations of antibodies, leading to a large financial and physical burden to AD patients. We hypothesize that scFvs against A¿ are effective and safe in treating AD patients as well as AD mouse models. We propose to use novel immune gene therapy for Alzheimer's disease, whereby recombinant adeno-associated virus (rAVV) vectors encoding anti-A¿ scFvs are directed to skeletal muscles in order to deliver the scFvs into the circulation and, probably, into the brain. This study will serve as a proof of principle to demonstrate if this gene therapy modality can deliver anti-A¿ scFvs into the circulation enough to reduce brain A¿ load and improve learning and memory deficits in an AD mouse model. In Specific Aim 1, we will evaluate prophylactic effects of muscle directed gene therapy modality by an rAAV vector encoding anti-A¿ scFv on prevention of A¿ deposits and behavioral deficits in an AD mouse model. In Aim 2, we will evaluate therapeutic effects of muscle-directed gene therapy modality by an rAAV vector encoding anti-A¿ scFv on clearance of A¿ deposits as well as improvement of behavioral deficits in an AD mouse model. We utilize neuropathological, biochemical, immunological, and behavioral analyses to determine the prophylactic and therapeutic effects and safety of the modality. The long-term goal is to establish the logical basis for developing safe and effective gene therapy modalities for AD utilizing rAAV and scFv.

描述（由申请人提供）：阿尔茨海默病（AD）是60岁以后痴呆症的最常见原因。阿尔茨海默病的病理特征包括淀粉样肽(A)在神经斑块和脑血管中的沉积、神经原纤维缠结和神经元的丧失。越来越多的证据支持这一观点，即A¿及其前体在阿尔茨海默病的发病机制中发挥重要作用。用合成A¿免疫阿尔茨海默病小鼠模型可减少A¿沉积并减轻其记忆和学习缺陷。然而，最近的临床试验因可能由t细胞介导和/或fc介导的免疫反应引起的脑炎症而暂停。在AD小鼠模型中，外周给药抗A¿抗体也诱导了预先存在的淀粉样斑块的清除，这表明活跃的t细胞介导的免疫反应是不必要的。在AD小鼠模型中，局部应用不含Fc抗体的F(ab)2可清除淀粉样蛋白沉积，这表明非Fc介导的机制也参与了清除。我们已经证明，抗A¿的人单链抗体（scFv）在AD小鼠模型中也能有效减少大脑A¿负荷。虽然这些被动免疫模式在治疗AD患者方面可能是有效的，而且不会引起炎症反应等副作用，但这种模式受到反复给药抗体的影响，给AD患者带来了巨大的经济和身体负担。我们假设抗A¿的scFvs在治疗AD患者和AD小鼠模型中是有效和安全的。我们建议使用新的免疫基因疗法治疗阿尔茨海默病，通过重组腺相关病毒（rAVV）载体编码抗-a¿scFvs，从而将scFvs传递到骨骼肌，从而将scFvs输送到血液循环中，甚至可能进入大脑。这项研究将作为原理证明，证明这种基因治疗方式是否可以将抗- a - scFvs输送到血液循环中，以减少大脑a -负荷，改善AD小鼠模型的学习和记忆缺陷。在Specific Aim 1中，我们将通过编码抗A¿scFv的rAAV载体评估肌肉定向基因治疗方式对AD小鼠模型中A¿沉积和行为缺陷的预防作用。在Aim 2中，我们将通过编码抗A¿scFv的rAAV载体评估肌肉定向基因治疗方式对AD小鼠模型中A¿沉积物的清除和行为缺陷的改善的治疗效果。我们利用神经病理学、生化、免疫学和行为分析来确定该方法的预防和治疗效果以及安全性。长期目标是为利用rAAV和scFv开发安全有效的AD基因治疗模式建立逻辑基础。