Pyroglutamate Amyloid-beta as an Immunotherapeutic Target for Alzheimer's Disease

焦谷氨酸淀粉样蛋白-β作为阿尔茨海默病的免疫治疗靶点

• 批准号: 8702980
• 负责人: CYNTHIA A LEMERE
• 金额: $ 33.89万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8702980
• 关键词: Acute; Affinity; Age; Age-Months; Aging; Alzheimer' s Disease; Amino Acids; Amyloid; Amyloid Proteins; Amyloid beta-Protein; Amyloid deposition; Animal Model; Antibodies; Behavioral; Bilateral; Binding; Biochemical; Blood Vessels; Brain; Canis familiaris; Carotid Artery Ulcerating Plaque; Cerebrum; Charge; Cognitive; Cognitive deficits; Collaborations; Cyclization; Data; Deposition; Disease Progression; Enzyme-Linked Immunosorbent Assay; Enzymes; Excision; Female; Flow Cytometry; Germany; Glutamic Acid; Goals; Human; Hybridomas; Image Cytometry; Immune response; Immunotherapeutic agent; Immunotherapy; Impaired cognition; In Vitro; Inflammation; Injection of therapeutic agent; Intraperitoneal Injections; Label; Learning; Left; Length; Life; Measures; Memory; Microglia; Monoclonal Antibodies; Mus; N-terminal; Neonatal; Nerve Degeneration; Neurodegenerative Disorders; Outcome Measure; Passive Immunization; Pathogenesis; Peptide antibodies; Peptides; Phagocytosis; Play; Prevention; Proteins; Pyroglutamate; Reagent; Reporting; Resistance; Role; Seeds; Staging; Technical Expertise; Testing; Therapeutic; Tissues; Transgenic Mice; Walkers; Water; aged; cellular imaging; cytokine; experience; extracellular; glutaminyl-peptide cyclotransferase; improved; in vivo; male; mouse model; neurotoxic; nonhuman primate; pathological aging; peptide A; prevent; response; synthetic peptide; therapeutic target; uptake

DESCRIPTION (provided by applicant): N-terminally-truncated and modified amyloid-beta (A¿) peptides are abundant in cerebral amyloid deposits in Alzheimer's disease (AD). Pyroglutamate A¿ is generated upon N-terminal truncation of A¿ followed by cyclization by glutaminyl cyclase to convert glutamic acid at residues 3 and 11 to pyroglutamate (A¿pE3 and A¿pE11). Both forms aggregate quickly, resist degradation, and are neurotoxic. It is unclear if either is present in initial A¿ deposition in plaques and blood vessels (i.e., acting as a seed for further deposition) or if they are modified later. However, Alzheimer's disease progression appears to correlate with the presence of A¿ pE peptide aggregates in brain. We hypothesize that pyroglutamate A¿ acts as a seed for A¿ deposition and accelerates inflammation, neurodegeneration and cognitive decline; therefore, targeted removal of this toxic species by immunotherapy will reduce A¿ deposition, inflammation and neuritic dystrophy, and protect against cognitive impairment without disturbing non-pathogenic A¿. We propose 4 Specific Aims. Aim 1: We will determine if intrahippocampal or intraperitoneal injections of A¿pE-containing mouse brain extracts enhance A¿ deposition, inflammation, neurodegeneration and cognitive decline in APP/PS1dE9 transgenic mice with aging in vivo. Aim 2: We will determine if early removal of pyroGlu A¿ prevents general A¿ plaque deposition and neuritic changes, and protects against cognitive decline by passively immunizing (i.p.) male APP/PS1dE9 tg mice with an anti-A¿N3pE mAb (07/1), an anti-A¿pE11 mAb, a general A¿ mAb (3A1), or PBS weekly from 4-12 mo of age, starting prior to plaque onset. Outcome measures: behavioral, biochemical, and neuropathological analyses. Aim 3: We will determine if removal of pyroGlu A¿ in late stage AD reduces total A¿ and neurodegeneration and improves cognitive deficits by passively immunizing (i.p.) female APP/PS1dE9 tg mice weekly from 12-16 mo of age, starting well after plaque onset. Antibodies and outcome measures are the same as in Aim 2. Aim 4: We will examine the immune response of microglia to pyroGlu A¿ mAbs in acute in vivo studies and in primary microglial cultures in vitro. Our collaborators at Probiodrug AG (Germany) will kindly provide us with 2 high-affinity, highly selective pyroGlu A¿ mAbs (anti- A¿ pE3 and anti-A¿pE11), synthetic A¿pE peptides, and brain extracts from their transgenic mouse models that accumulate pyroGlu-3 A¿ peptides. Our collaborators at the CND have generously provided the 3A1 general A¿ mAb hybridoma as a control. Importantly, my lab initiated this collaboration and has many years of experience investigating pyroGlu A¿ deposition, inflammation, and A¿ immunotherapy. The overall goal of our study is to determine whether pyroglutamate A¿ proteins (A¿pE3 and A¿pE11) are therapeutic targets for Alzheimer's disease and, whether clearance by immunotherapy specific for either pyroGlu A¿ species would be efficacious to prevent and/or treat AD.

描述（由适用提供）：在阿尔茨海默氏病（AD）的脑淀粉样蛋白沉积中，N末端截短和改良的淀粉样蛋白β（A»）肽丰富。焦谷氨酸a是在A检授的N末端截断后生成的，然后通过谷氨酰胺基环酶环化，将3和11残留的谷氨酸转化为焦谷氨酸（a¿pe3和a¿pe11）。两种形式都会迅速聚集，抵抗降解，并且是神经毒性的。目前尚不清楚在斑块和血管中的初始A沉积中是否存在任何一个（即，作为种子的种子 进一步的沉积）或以后进行修改。然而，阿尔茨海默氏病的进展似乎与大脑中A Peperped骨料的存在相关。我们假设焦谷氨酸a已成为沉积物的种子，并加速了感染，神经退行性和认知能力下降。因此，通过免疫疗法靶向去除这种有毒物种将减少沉积，感染和神经性营养不良，并防止不干扰非致病性A。的认知障碍。我们提出了4个具体目标。 AIM 1：我们将确定对含Pe的小鼠脑提取物的汉皮门内或腹膜内注射是否会增强APP/PS1DE9转基因小鼠的沉积，炎症，神经变性和认知能力下降。目的2：我们将确定早期清除焦图是否可以防止一般A斑块沉积和神经元变化，并通过被动免疫（I.P.）App/ps1de9 tg小鼠防止认知下降，并使用抗A an 33pe mab（07/1），抗A anti-a¿年龄，从牙菌斑发作之前开始。结果指标：行为，生化和神经病理学分析。 AIM 3：我们将确定在晚期AD中除去焦点a。是否可以通过被动免疫（I.P.）女性App/PS1DE9 TG小鼠每周从12-16 MO降低了总体A和神经变性，并改善了认知缺陷，从12-16 Mo开始，在斑块发作后开始良好。抗体和结局指标与AIM 2相同。目标4：我们将检查小胶质细胞对急性体内研究中的小胶质细胞的免疫响应，并在体内研究和体外原发性小胶质细胞培养中。我们在Pribiodrug AG（德国）的合作者将为我们提供2种高亲和力，高度选择性的pyroglua¿mabs（抗a pe3和抗A pe11），合成pe肽，大脑从其转基因小鼠模型中提取pyroglu-3 a？pypypepides。我们在CND的合作者慷慨地为3A1 General提供了MAB杂交瘤作为对照。重要的是，我的实验室启动了这一合作，并具有多年的研究经验，这些经验研究了焦图，炎症和免疫疗法。我们研究的总体目的是确定焦谷氨酸A蛋白（a¿pe3和a¿pe11）是否是阿尔茨海默氏病的治疗靶标，以及是否有效地通过对任何一种pyroglu物种进行特定的免疫疗法来有效地预防和/或治疗AD。