Development of Novel Synthetic Proteomimetics for Mediating Tauopathy in Alzheimer's Disease

项目总结 阿尔茨海默病(AD)是一种无法治愈的进行性神经退行性疾病，严重影响3500万人 世界各地患有痴呆症和认知衰退的人。最近，微管相关蛋白Tau 被认为是阿尔茨海默病的重要诱因，因为它的错误折叠和聚集破坏了动态平衡和 导致广泛的神经元功能障碍。目前封顶Tau的努力仍然面临重大的临床挑战 来自生长或降解大缠结的聚集体：a)小分子方法具有非特异性结合 和由此产生的副作用，b)抗体触发免疫反应，需要对每个抗体进行修饰 患者，以及c)有希望的牛磺酸结合肽具有较差的细胞渗透性并且容易被降解。要克服 这些挑战，这项建议利用了一种新的方法，即类蛋白质聚合物(PLP)来保护Tau 用于持续递送的结合多肽的降解以防止和封堵原纤维的形成和 使用蛋白质水解靶向嵌合体(PROTAC)PLP方法降低较大聚集体的扩散。这 合成蛋白质组学将多肽包装在一起，作为高密度刷状聚合物，是模块化的，可扩展的， 并使用现代先进的聚合策略快速配制，以提供多价性、细胞渗透性、 和特定的靶标结合。与小分子相比，PLP表现出更长的循环半衰期和线性 多肽，维持强大的生物活性、细胞摄取和破坏蛋白质-蛋白质相互作用(PPI)。这里, 我们的目标是通过两种方式来靶向Tau：1.选择性地与聚集前的Tau结合以抑制聚集，以及2. 为了提高PROTACs酶的稳定性和交付能力，采用一种新的蛋白质仿生生物偶联策略。 利用开环歧化聚合(ROMP)技术，我将开发和优化一个Tau结合文库 PLPS和Tau PROTAC PLPS。所有的PLP都将在体外降解中进行严格的表征和评估 和稳定性研究，以及在相关Tau表达细胞中的疗效。我会直接比较一下表现 Tau PLP在细胞活力、细胞摄取、Tau结合以及对Tau聚集体形成的影响 天然多肽及其相应的线性PROTAC。这将与我的共同赞助人合作完成， 蛋白质病分子生物学专家理查德·森本教授说。我和我的赞助商/共同赞助商 我在西北大学设计了一个培训计划，以发展我作为化学生物学博士的独立研究技能 候选人。我将在西北大学进行这项跨学科的研究，利用 为这两个项目开发技能的核心设施(疾病细胞模型、化学合成、高级成像) 以及我未来在神经化学领域的独立研究生涯。我会同时提高我的口语和 书面科学交流，在专业发展研讨会上建立网络，并影响我的 社区。在教授的指导下Gianneschi和Morimoto，我将培养成为不可或缺的纳米医学的技能 研究员。在Kirchstein NRSA的支持下，我将识别和优化一种化学生物学工具或纳米疗法 在阿尔茨海默氏症中调节调节失调的Tau，以减轻我们世界老龄化人口的发病率。

PROJECT SUMMARY Alzheimer’s Disease (AD) is an incurable, progressive neurodegenerative disease drastically affecting 35 million people worldwide who suffer from dementia and cognitive decline. Recently, microtubule associated protein Tau is believed to be an instrumental inducer in AD, as its misfolding and aggregation disrupts homeostasis and results in widespread neuronal dysfunction. Significant clinical challenges remain in current efforts to cap Tau aggregates from growing or degrading large tangles: a) small molecule approaches have nonspecific binding and resultant side effects, b) antibodies trigger an immune response that necessitates modification for each patient, and c) promising Tau binding peptides have poor cell permeability and are easily degraded. To overcome these challenges, this proposal utilizes a new methodology, the Protein-Like Polymer (PLP) for protecting Tau binding peptides from degradation for their sustained delivery to prevent and cap protofibril formation and degrade larger aggregates from spreading using a proteolysis targeting chimera (PROTAC) PLP approach. This synthetic proteomimetic packages peptides together as high-density brush polymers that are modular, scalable, and rapidly formulated using modern, advanced polymerization strategies to offer multivalency, cell penetration, and specific target binding. PLPs exhibit extended circulation half-life compared to small molecules and linear peptides, maintaining strong bioactivity, cell uptake, and disrupting protein-protein interactions (PPIs). Here, we aim to target Tau in two ways: 1. selectively binding to pre-aggregative Tau to dampen aggregation, and 2. to improve PROTACs enzyme stability and deliverability using a novel proteomimetic bioconjugation strategy. Using Ring Opening Metathesis Polymerization (ROMP), I will develop and optimize a library of Tau binding PLPs and Tau PROTAC PLPS. All PLPs will be rigorously characterized and assessed in in vitro degradation and stability studies, as well as efficacy in relevant Tau expressing cells. I will directly compare the performance of the Tau PLPs in cell viability, cellular uptake, Tau binding, and effect on Tau aggregate formations to the native peptides and their corresponding linear PROTACs. This will be done in collaboration with my co-sponsor, Prof. Richard Morimoto, an expert in proteinopathy molecular biology. My sponsor/co-sponsor and I have devised a training plan at Northwestern to develop my independent resesarch skills as a Chemical Biology PhD candidate. I will conduct this interdisciplinary research at Northwestern, taking advantage of technical training at core facilities to develop skills for both this project (disease cell models, chemical synthesis, advanced imaging) and my future independent research career in neurochemistry. I will simultaneously be improving my oral and written science communication, networking at professional development workshops, and impacting my community. Under Profs. Gianneschi and Morimoto, I will cultivate the skills to be an indispensable nanomedicine researcher. With Kirchstein NRSA Support, I will identify and optimize a chemical biology tool or nanotherapeutic for mediating dysregulated Tau in Alzheimer’s Disease to allay the morbidity of our world’s aging population.