Development of A Novel Class of Protein Conformation Selective Molecular Sensors

PI：CHI，EvaProposal No：1605225该项目将开发一种新型传感器，用于早期检测和跟踪阿尔茨海默氏症和帕金森氏症等神经退行性疾病。特别是，传感器将检测到早在症状出现前20年就在大脑中形成的蛋白质聚集体。检测和跟踪这些蛋白质聚集体的能力不仅有助于更好地了解疾病，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治疗方法。蛋白质的错误折叠和聚集是阿尔茨海默病和帕金森病等神经退行性疾病的中心致病事件，特别是神经毒性最强的纤维前聚集体构象，这可能是由于缺乏能够选择性和差异性靶向不同蛋白质聚集体构象的分子探针。为了满足这一迫切的需求，开发了一种新型的寡聚(对苯乙炔)电解质(OPES)，用于基于多种“荧光开启”机制直接传感广泛的淀粉样聚集体。与现有的临床使用有限的探针(如基于硫代黄素-T的探针)相比，Opes具有许多独特的优势，包括多功能性和高度可定制的结构和化学性质，最显著的是Opes对与配体相互作用的多种反应模式。具体地说，研究人员将合成并测试OPES，用于体外检测阿尔茨海默氏症和帕金森氏症相关蛋白制备的蛋白质聚集体(目标1)。实验结果还将与密切相关的计算建模相结合，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。这些见解将被用来指导OPES的合理设计和合成，体外检测脑组织切片中的淀粉样聚集体(目标2)和活体检测阿尔茨海默病动物模型脑中的淀粉样聚集体(目标3)。该项目开发的分子传感器将为研究人员提供同时动态跟踪体外和体内系统蛋白质错误折叠和聚集过程的工具，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，拟议的多学科研究将使私人投资机构有义务培训研究生和本科生学习现代方法，以解决化学、生物、工程和医学之间的重要问题。所获得的跨学科技能将为学生在学术界、国家实验室和工业领域的职业生涯做好准备。PIS还提出了一项雄心勃勃的计划，即设计生物传感器设计挑战模块，并将其纳入新墨西哥大学各级核心化学工程本科课程。这一教育努力的目标有两个：1.提高学生的留存率，特别是代表人数较少的少数族裔；2.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.