Mosaic Display of Multivalent Tau and A-Beta peptides on Immunogenic SNAP Liposomes

项目概要/摘要 阿尔茨海默氏病（AD）目前影响约620万美国人，预计这一数字将增加到 到2060年将达到1400万，除非发现预防或延迟AD发作的新治疗或干预措施。 利用免疫系统来预防或去除Aβ和/或tau病理是一种有希望的疾病- 改变治疗AD的治疗方法。目前，所有以前和正在进行的免疫治疗 研究仅针对Aβ或tau蛋白的一个表位。因为大多数AD病例是多因素的， 由于散发性AD是由多种机制引起的，因此单个靶点不太可能足以阻止或减少 AD的进展。基于Aβ和tau蛋白在AD发病机制中的作用， 同时靶向Aβ和tau的多个表位可能为AD药物提供有希望的方法 发展拟议项目的长期目标是开发一种新型有效的AD疫苗。 该项目的具体目标是开发一种多价“马赛克”疫苗，称为SNAP-AD，靶向多个 Aβ和tau的表位同时与自发纳米脂质体抗原颗粒（SNAP）平台， 研究其在转基因小鼠模型中治疗AD的疗效。我们的假设是 同时靶向Aβ和tau的多个表位的免疫疗法是一种有前途的方法， 开发有效的AD治疗剂。为了验证这一新的假设，我们提出了三个具体目标：（1） 配制并表征SNAP-AD，一种包含三个tau和两个Aβ肽的镶嵌纳米颗粒疫苗 表位POP BIO将同时针对SNAP-AD五价疫苗进行配制和表征 在一个N-末端区域（Tau 6-18）、一个中间区域（Tau 184-195）和一个C-末端区域（pS396/S404）处的tau， 以及N-末端Aβ（1-14）和N-末端焦谷氨酸Aβ（pE 3 -14），并开发了制备方法 具有可重复的批次间表征和定量每种肽和免疫原性的方案， 脂质成分(2)评估SNAP-AD的免疫干扰、Th 1偏向性细胞应答， 神经炎症和抗体的持久性。将在5重中评估免疫干扰 免疫原，如果观察到，将测试可能的衍生物方法。细胞应答，Th 1/Th 2偏倚 将探索由SNAP-AD诱导的抗体应答和抗体的持久性。(3)确定SNAP- 使用3xTg-AD小鼠模型的AD功效。10月龄3xTg-AD小鼠和年龄匹配的野生型小鼠 将用SNAP-AD免疫接种，并以四周间隔加强两次。第三剂后一个月 免疫，行为测试，包括一般行为，焦虑，运动活动和认知功能， 将进行生物化学/生物化学研究，并研究Aβ和tau病理学。的 提出的研究将揭示多价免疫疗法是否具有有希望的治疗效果 AD病理学和认知障碍的研究这个项目的积极成果可能会导致一部小说 用于治疗AD的疫苗，这将改善AD患者的生活质量。

Project Summary/Abstract Alzheimer’s disease (AD) currently affects ~6.2 million Americans, and this number is projected to increase to 14 million by 2060 unless novel treatments or interventions to prevent or delay the onset of AD are identified. Harnessing the immune system to prevent or remove Aβ and/or tau pathologies represents a promising disease- modifying therapeutic approach for the treatment of AD. Currently, all the previous and ongoing immunotherapy studies target only one epitope of either Aβ or tau protein. Because most AD cases are multifactorial, and sporadic AD is caused by multiple mechanisms, it is unlikely that a single target will be sufficient to stop or reduce the progression of AD. Based on the role of Aβ and tau in the pathogenesis of AD, immunotherapy simultaneously targeting multiple epitopes of both Aβ and tau may present a promising approach for AD drug development. The long-term objective of the proposed project is to develop a novel, effective vaccine for AD. The specific goal of this project is to develop a multivalent "mosaic" vaccine, termed SNAP-AD, targeting multiple epitopes of Aβ and tau simultaneously with the spontaneous nanoliposome antigen particle (SNAP) platform and to investigate its therapeutic effects for treating AD in a transgenic mouse model. Our working hypothesis is that immunotherapy simultaneously targeting the multiple epitopes of both Aβ and tau is a promising approach for developing effective AD therapeutics. To test this novel hypothesis, we propose three specific aims: (1) Formulate and characterize SNAP-AD, a mosaic nanoparticle vaccine comprising three tau and two Aβ peptide epitopes. POP BIO will formulate and characterize the pentavalent SNAP-AD vaccine simultaneously targeting tau at one N-terminal region (Tau 6-18), one mid-region (Tau 184-195), and one C-terminal region (pS396/S404), as well as N-terminal Aβ (1-14) and N-terminal pyroglutamate Aβ (pE3-14), and develop a manufacturing method with reproducible batch-to-batch characterization and the protocols to quantify each peptide and immunogenic lipid component. (2) Assess SNAP-AD for immune interference, Th1-biased cellular response, neuroinflammation, and antibodies’ durability. Immune interference will be assessed amongst the 5-plex immunogens, and if observed, possible ramification methods will be tested. Cellular responses, Th1/Th2 bias of the antibody response, and the antibodies’ durability induced by SNAP-AD will be probed. (3) Determine SNAP- AD efficacy using the 3xTg-AD mouse model. Ten-month-old 3xTg-AD mice and age-matched wild-type mice will be immunized with SNAP-AD and boosted twice with a four-week interval. One month after the third dose of immunization, behavioral tests, including general behavior, anxiety, locomotor activity, and cognitive function, will be conducted, and Aβ and tau pathology will be investigated both biochemically/immunohistochemically. The proposed studies will reveal whether the multivalent immunotherapy approach has promising therapeutic effects on AD pathologies and cognitive impairment. A positive outcome of this project would potentially lead to a novel vaccine for the treatment of AD, which will improve the quality of life of individuals with AD.