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

项目摘要/摘要 阿尔茨海默病(AD)目前影响着约620万美国人，预计这一数字将增加到 到2060年达到1400万人，除非找到预防或延缓阿尔茨海默病发病的新疗法或干预措施。 利用免疫系统来预防或消除β和/或tau病变是一种很有前途的疾病- 改良治疗阿尔茨海默病的方法。目前，所有先前和正在进行的免疫治疗 研究只针对Aβ或tau蛋白的一个表位。因为大多数AD病例是多因素的，而且 散发性AD是由多种机制引起的，单个目标不太可能足以阻止或减少 AD的进展。基于Aβ和tau在AD发病机制中的作用，免疫治疗 同时靶向Aβ和tau的多个表位可能为AD药物提供一种有前途的方法 发展。拟议项目的长期目标是开发一种新的、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，针对多个 Aβ和tau表位同时与自发纳米脂质体抗原颗粒平台和 目的：探讨其对转基因小鼠阿尔茨海默病的治疗作用。我们的工作假设是 同时靶向Aβ和tau多个表位的免疫治疗是一种很有前途的治疗方法 开发有效的阿尔茨海默病疗法。为了验证这一新的假设，我们提出了三个具体目标：(1) 含三个tau和两个Aβ多肽的嵌合型纳米疫苗SNAP-AD的研制与鉴定 表位。POP BIO将制定和表征同时靶向的五价SNAP-AD疫苗 位于一个N-末端区域(Tau 6-18)、一个中间区(Tau 184-195)和一个C-末端区域(pS396/S404)的tau， 以及N端Aβ(1-14)和N端焦谷氨酸盐Aβ(PE3-14)，并开发了一种制造方法 具有可重复的批次到批次的表征和用于量化每个多肽和免疫原性的方案 脂类成分。(2)评估SNAP-AD的免疫干扰、偏向Th1的细胞反应、 神经炎症和抗体的持久性。免疫干扰将在5-plex中进行评估 免疫原，如果观察到，可能的分支方法将进行测试。细胞反应，Th1/Th2偏向 并对SNAP-AD诱导的抗体应答和抗体持久性进行了探讨。(3)确定SNAP- 采用3xTg-AD小鼠模型观察AD疗效。10月龄3xTg-AD小鼠和年龄匹配的野生型小鼠 将使用SNAP-AD进行免疫，并以四周为间隔加强两次。第三次服药后一个月 免疫、行为测试，包括一般行为、焦虑、活动和认知功能， 将进行，Aβ和tau病理将进行生化/免疫组化研究。这个 拟议的研究将揭示多价免疫疗法是否具有良好的治疗效果。 关于阿尔茨海默病的病理和认知障碍。这个项目的积极结果可能会导致一部小说 治疗阿尔茨海默病的疫苗，这将改善阿尔茨海默病患者的生活质量。

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.