Peptoid Antagonists of the ApoE/ABeta Interaction as a Novel Anti-ABeta Therapy.

项目总结： 这是K02 AG034176独立科学家职业发展奖的行政补充申请，该奖项受到超级风暴桑迪的影响。该奖项的研究计划致力于开发阻断Aü和载脂蛋白(Apo)E之间相互作用的类肽化合物，作为阿尔茨海默病(AD)的新疗法。职业发展计划概述了蛋白质化学、肽类化合物合成、药物设计和开发、体内微透析、获得进行AD临床试验的经验以及指导初级教员和博士生的培训。由于纽约大学医学院校区持续的洪水，我们的多个研究设施暂时关闭，一些仍然没有重新开放，这对校园的学术和培训活动造成了影响。暂时关闭包括我们的实验室和动物群体设施，那里被洪水淹没，五周内无法进入；因此，所有培育AD转基因(TG)小鼠以进行计划实验和正在进行的这些小鼠实验的努力都被暂停。尽管遭遇挫折，但该奖项的具体目标保持不变。目的概述apoE/A？结合的类肽拮抗剂的研究进展，它们是基于A？12-28p的结构设计的，A？12-28p是我们的初步研究中使用的一种合成肽，目的是证明药物阻断apoE/A？相互作用可防止记忆障碍并减少AD TG小鼠的A？斑块负荷(Sadowski等人)。PNAS2006年；第103：18787页)。目的II概述A？12-28P及其最适肽样衍生物对表达不同人载脂蛋白E亚型的AD TG小鼠的治疗，作为对表达小鼠载脂蛋白E的小鼠的初步研究。目的III研究阻断apoE/A？相互作用是否能阻止A？神经元内蓄积，改善脑内A？清除。本增刊的研究计划建立在朝着最初的具体目标努力取得的结果的基础上，并提出了恢复超级风暴桑迪造成的损失的研究。对于Aim I，我们有一个独特的带有肽类修饰的环状A？12-28p衍生物的文库，我们建议对其进行测试。对于AIM II，我们开发并鉴定了一种新的AD TG模型小鼠APPSW/PS1dE9 人载脂蛋白E亚型E2、E3和E4的小鼠载脂蛋白E基因的靶向替换的APOE-TR。在APPSW/PS1dE9/APOE2-tr和APPSW/PS1dE9/apoE4-tr小鼠中，A？12-28P处理都减少了脑A？蓄积。在本补充材料中，我们计划在APPSW/PS1dE9/apoE3-tr品系中描述A？12-28P处理的特征，以及在所有三个tr品系中使用最有希望的类肽化合物处理的效果。对于Aim III，我们建立了一个神经元-星形胶质细胞共培养系统来研究apoE/A？的体外相互作用，并计划研究不同的人类apoE亚型对神经元内A？病理的影响。此外，这一行政补充将有助于恢复我最初的职业发展计划，该计划在超级风暴过后被打乱，原因是我们校园的多个设施关闭，以及需要将个人精力从职业发展和培训活动转移到恢复我们实验室的研究能力上。

Project summary: This is an application for an administrative supplement to K02 AG034176 Independent Scientist Career Development Award, which was affected by the super storm Sandy. The research plan of this award pursues development of peptoid compounds blocking the interaction between Aß and apolipoprotein (apo) E as novel Alzheimer’s disease (AD) therapeutics. The career development plan outlines training in protein chemistry, synthesis of peptoid compounds, drug design and development, in vivo Aß microdialysis, gaining experience in conducting AD clinical trials, and mentoring junior faculty and PhD students. Due to flooding sustained by the NYU School Medicine campus multiple of our research facilities were temporarily closed and some still have not reopened, which compromised on campus academic and training activities. Temporary closure included our laboratory and our animal colony facility, which was flooded and became inaccessible for five weeks; therefore all efforts to breed AD transgenic (Tg) mice for planned experiments and ongoing experiments on these mice were suspended. Despite setbacks, the specific aims of the award remained unchanged. Aim I outlines development of peptoid antagonists of the apoE/Aß binding, which are designed based on the structure of Aß12- 28P, a synthetic peptide used in our preliminary study to demonstrate that pharmacological blocking of the apoE/Aß interaction prevents memory impairment and reduces Aßplaque load in AD Tg mice (Sadowski et al. PNAS 2006;103:18787). Aim II outlines treatment with Aß12-28P and its most optimal peptoid derivatives in AD Tg mice expressing various human apoE isoforms, as the preliminary study was done in mice expressing murine apoE. Aim III investigates whether blocking the apoE/Aß interaction prevents intraneuronal accumulation of Aß and improves brain Aß clearance. The research plan of this supplement builds on results, which were obtained working toward the original specific aims and proposes studies to restore losses caused by the super storm Sandy. For aim I we have a unique library of cyclic Aß12-28P derivatives with peptoid modifications which we propose to test. For aim II we have developed and characterized a novel AD Tg model mice APPSW/PS1dE9 apoE-TR with targeted replacement (TR) of the mouse apoE gene for human apoE isoforms E2, E3, and E4. Aß12-28P treatment in both APPSW/PS1dE9/apoE2-TR and APPSW/PS1dE9/apoE4-TR mice reduced brain Aß accumulation. In this supplement we plan to characterize Aß12-28P treatment in APPSW/PS1dE9/apoE3-TR line and effects of the treatment with the most promising peptoid compound in all three TR lines. For aim III we have developed a neuronal-astrocytic co-culture system to study the apoE/Aß interaction in vitro and we plan to investigate the effect of various human apoE isoforms on intraneuronal Aß pathology. Furthermore, this administrative supplement would aid to restore my original career development plan, which was disrupted in the aftermath of the super storm due to closure of multiple facilities in our campus and a need to divert personal effort from career development and training activities toward restoration of research capacity of our laboratory.