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

项目摘要 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 全世界患有痴呆症和认知能力下降的人。最近，微管相关蛋白Tau 被认为是AD中的工具诱导物，因为其错误折叠和聚集破坏体内平衡， 导致广泛的神经元功能障碍。目前限制Tau的努力仍面临重大临床挑战 生长或降解大缠结产生的聚集体：a）小分子方法具有非特异性结合 和由此产生的副作用，B）抗体触发免疫应答， c）有希望的Tau结合肽具有差的细胞渗透性并且容易降解。克服 面对这些挑战，该提案利用了一种新的方法，即蛋白质样聚合物（PLP）来保护Tau 结合肽免于降解，用于它们的持续递送以防止和覆盖原纤维形成， 使用蛋白质水解靶向嵌合体（PROTAC）PLP方法降解散布的较大聚集体。这 合成的蛋白质模拟物将肽包装在一起作为高密度刷状聚合物，其是模块化的，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，细胞渗透， 和特异性靶结合。PLP与小分子和线性PLP相比，表现出延长的循环半衰期。 肽，保持强的生物活性，细胞摄取，并破坏蛋白质-蛋白质相互作用（PPI）。在这里， 我们的目标是以两种方式瞄准Tau：1.选择性结合预聚集Tau以抑制聚集，和2. 使用新的蛋白质模拟物生物缀合策略来改善PROTAC酶的稳定性和递送能力。 使用开环易位聚合（ROMP），我将开发和优化Tau结合文库， PLPs和Tau PROTAC PLPS。所有PLP将在体外降解中进行严格表征和评估 和稳定性研究，以及在相关Tau表达细胞中的功效。我会直接比较性能 在细胞活力、细胞摄取、Tau结合和对Tau聚集体形成的影响方面， 天然肽及其相应的线性PROTAC。这将与我的共同赞助者合作进行， 教授理查德·森本，蛋白质病分子生物学专家。我的赞助商/共同赞助商和我有 作为一名化学生物学博士，我在西北大学设计了一个培训计划，以发展我的独立研究技能 候选人我将在西北大学进行这项跨学科研究，利用技术培训， 核心设施，以发展技能，这两个项目（疾病细胞模型，化学合成，先进的成像） 以及我未来在神经化学方面的独立研究事业我将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Profs. Gianneschi和Morimoto，我将培养的技能是不可或缺的纳米医学 研究员通过Kirchstein NRSA支持，我将识别和优化化学生物学工具或纳米技术 在阿尔茨海默病中介导Tau失调，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.