Development of A Novel Class of Protein Conformation Selective Molecular Sensors

该项目将开发一种新型传感器，用于早期检测和跟踪神经退行性疾病，如阿尔茨海默病和帕金森病。特别是，这些传感器可以在症状出现前20年就检测到大脑中形成的蛋白质聚集体。检测和追踪这些蛋白质聚集体的能力不仅有助于更好地了解这些疾病，而且还将导致早期诊断，并有助于开发预防和治疗这些毁灭性疾病的疗法。蛋白质的错误折叠和聚集是神经退行性疾病（如阿尔茨海默病和帕金森病）的主要致病事件，特别是最具神经毒性的纤维前聚集构象，这可能是由于缺乏能够选择性和差异地靶向不同蛋白质聚集构象的分子探针。为了满足这一关键需求，提出了一种基于多种“荧光开启”机制的新型寡聚（对苯基乙炔）电解质（OPEs）的开发，用于直接传感广泛的淀粉样蛋白聚集体。与现有的临床应用有限的探针相比，例如基于硫黄素- t的探针，OPEs具有许多明显的优势，包括通用和高度可定制的结构和化学性质，最值得注意的是OPEs响应与配体相互作用的多种模式。具体而言，研究人员将合成并测试OPEs，用于阿尔茨海默病和帕金森病相关蛋白制备的蛋白聚集体的体外检测（目的1）。实验结果也将与密切相关的计算模型协同结合，以获得对OPE-蛋白质聚集相互作用以及OPE的传感机制的基本理解。这些见解将用于指导OPEs的合理设计和合成，脑组织切片淀粉样蛋白聚集体的体外检测（目标2）和阿尔茨海默病活体动物模型脑的体内检测（目标3）。该项目开发的分子传感器将为研究人员提供工具，同时动态跟踪体外和体内系统的蛋白质错误折叠和聚集过程，促进对主要神经退行性疾病的病因、诊断和治疗的研究。此外，拟议中的多学科研究将要求pi培养研究生和本科生掌握处理化学、生物、工程和医学之间的重要问题所需的现代方法。所获得的跨学科技能将为学生在学术、国家实验室和工业领域的职业生涯做好准备。pi还提出了一项雄心勃勃的计划，将“生物传感器设计挑战模块”设计并整合到新墨西哥大学各级化学工程本科核心课程中。这一教育努力的目标是双重的：1。提高学生留存率，尤其是少数族裔学生；通过在整个本科教育中提供解决开放式设计问题的机会，提高学生的成功。

PI: Chi, EvaProposal No: 1605225This project will develop a novel class of sensors for the early detection and tracking of neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. In particular, the sensors will detect the protein aggregates that form in the brain as early as two decades before the onset of symptoms. The ability to detect and track these protein aggregates will not only help in understanding the diseases better, but will also lead to early diagnosis and contribute towards the development of therapies to prevent and treat these devastating diseases. Misfolding and aggregation of proteins is a central pathogenic event in neurodegenerative disorders such as Alzheimer's and Parkinson's diseases, particularly the pre-fibrillar aggregate conformations that are the most neurotoxic, which is potentially due to a lack of molecular probes that could selectively and differentially target different protein aggregate conformations. To meet this critical need, development of a novel oligo(p-phenylene ethynylene) electrolytes (OPEs) for the direct sensing of the wide set of amyloid aggregates based on multiple "fluorescence turn-on" mechanisms is proposed. Compared to existing probes that are of limited clinical use, e.g., Thioflavin-T based probes, OPEs offer many distinct advantages, including versatile and highly tailorable structural and chemical properties, and most notably the multiple modes by which OPEs respond to interactions with ligands. Specifically, the researchers will synthesize and test OPEs for the in vitro detection of protein aggregates prepared from Alzheimer's and Parkinson's associated proteins (Objective 1). Experimental findings will also be synergistically combined with closely related computational modeling to gain a fundamental understanding of OPE-protein aggregate interactions as well as OPE's sensing mechanism. Such insights will be used to guide the rational design and synthesis of OPEs Ex vivo detection of amyloid aggregates in brain tissue sections (Objective 2) and in vivo detection in live Alzheimer's animal model brains (Objective 3). Molecular sensors developed in this project will give researchers the tool to simultaneously and dynamically track the protein misfolding and aggregation process for both in vitro and in vivo systems, facilitating research into the cause, diagnosis, and treatment of major neurodegenerative disorders. Additionally, the proposed multidisciplinary research will obligate the PIs to train graduate and undergraduates students in modern methodologies required to address important problems at the interface between chemistry, biology, engineering, and medicine. The acquired interdisciplinary skills will prepare students for careers in academe, national laboratories, and industry. The PIs also propose an ambitious plan to design and integrate "Biosensor Design Challenge Modules" into the core chemical engineering undergraduate curriculum at the University of New Mexico at all levels. The goal of this educational effort is two fold: 1. Improving student retention, particularly underrepresented minorities, and 2. Enhancing student success by providing opportunities for solving open-ended design problems throughout undergraduate education.