Screening and Immunization: A Novel Approach to Tau Antibody Therapeutics

描述(由申请人提供)：目前约有530万美国人患有阿尔茨海默病，在缺乏有效治疗的情况下，预计到2050年，这一数字将激增至1400万以上。目前针对A？A的免疫研究？病理学在稳定或恢复认知缺陷方面基本上没有成功，这表明它们没有针对合适的物种：tau。当患者表现出痴呆症的迹象时，tau病理可能是疾病的主要驱动因素，而不是A？目前tau抗体治疗策略的一个主要限制是依赖于线性表位结合。 用于筛选、选择和生产新型抗体。以这种方式，与生理相关的tau物种可能不会成为目标。我的中心假设是，可以通过两种不同的方法来改进抗tau抗体的开发领域：通过使用新的筛选技术来选择单抗，通过优化免疫抗原来生产单抗。这项建议的主要目标是：1)确定体外阻断tau接种的效果与体内功能恢复之间是否存在相关性，如钻石和Holtzman实验室(Yanamandra等人，正在出版)最近的一项研究所建议的那样；2)研究用于动物免疫的新型tau聚集体。目的1检验体外抑制tau接种是否能预测体内疗效。将使用两种不同的聚集检测方法对一组预先存在的抗体进行筛选，以确定它们抑制tau种子的能力。将在一系列有效性中选择六种抗体，用于体内注射中老年P301S小鼠。在6周或18周后 在治疗过程中，将对动物进行病理学(免疫组织化学和ELISA)、行为(恐惧条件作用)和突触(功能连接和光学内在信号成像)的评估，以确定tau种子抑制是否预示着功能恢复。目标2将从一组tau抗原中开发出新的抗tau抗体。不同大小的重组或人源性tau聚集体(大纤维、三聚体、~10聚体、~20聚体、~40聚体)将用于免疫小鼠，并对所产生的抗体进行抗原结合能力和tau种子抑制的筛选。对于每个抗原，将选择六种抗体进行纯化，并在本提案范围以外的未来研究中使用。总体而言，由于我们针对tau种子或种子活性的创新方法，而不是线性表位，预计这项提议将产生比目前可用的抗体更好的抗体。这些目标的实现将对未来AD新疗法的发展产生重大而直接的影响。

DESCRIPTION (provided by applicant): Approximately 5.3 million Americas currently suffer from Alzheimer's disease and, in the absence of effective treatments, this number is expected to balloon to over 14 million by the year 2050. Current immunization studies targeting A? pathology have been largely unsuccessful at stabilizing or restoring cognitive deficits, suggesting that they do not target the proper species: tau. By the time patients are showing signs of dementia, tau pathology is likely to be the principal driver of disease, not A?. A major limitation to current tau antibody therapeutic strategies is the reliance on linear epitope binding for screening, selection, and production of novel antibodies. In this manner, physiologically-relevant tau species may not be targeted. My central hypothesis is that the field of anti-tau antibody development can be improved by two separate means: by employing novel screening technology for selection of monoclonal antibodies and by optimizing immunization antigens for production of monoclonal antibodies. The main goals of this proposal are 1) to determine whether a correlation exists between efficacy of blocking tau seeding in vitro and functional recovery in vivo, as suggested in a recent study by the Diamond and Holtzman Laboratories (Yanamandra et al., in press) and 2) to investigate novel tau aggregates for animal immunization. Aim 1 will test if inhibition of tau seeding in vitro predicts in vivo efficacy. A lage panel of pre-existing antibodies will be screened for their capacity to inhibit tau seeding using two separate aggregation detection assays. Six antibodies will be chosen, over a range of efficacies, for in vivo administration into mid- to late-age P301S mice. After either 6 or 18 weeks of treatment, animals will be assessed pathologically (with immunohistochemistry and ELISA), behaviorally (with fear conditioning), and synaptically (with functional connectivity and optical intrinsic signal imaging) to determine if tau seeding inhibition is predictive of functional recovey. Aim 2 will develop novel anti-tau antibodies from a panel of tau antigens. Recombinant or human-derived tau aggregates of variable sizes (large fibrils, trimer, ~10mer, ~20mer, ~40mer) will be used to immunize mice, and resultant antibodies will be screened for antigen-binding capacity and tau seeding inhibition. Per antigen, six antibodies will be selected for purification and use in future studies outside the scope of this proposal. Cumulatively, it is expected that thi proposal will produce antibodies superior to those currently available because of our innovative approach for targeting tau seeds or seeding activity, rather than linear epitopes. The completion of these Aims will have a significant and direct impact on the development of future novel AD therapeutics.