CAREER: Quantitative Tools to Measure the Spatiotemporal Dynamics of Protein Signaling Complexes

尽管美国有超过1000万人患有学习和记忆障碍，但其中许多障碍的根本原因仍不清楚。这些疾病大多是由于蛋白质网络（称为蛋白质信号复合物）的调节中断而引起的，蛋白质网络控制着神经元相互交流的连接（称为突触）的强度。利用复杂的计算机建模和超分辨率显微镜，该项目寻求在时间和空间上对这些关键蛋白质复合物的动态组织产生重要的新分子水平理解。这种理解可能会导致设计新的治疗学习和记忆障碍的方法。此外，计划中的研究将为当前和未来的生物医学工程专业学生提供进入快速发展的生物技术和生物医学工程领域所需的多学科培训。通过参与跨学科工程设计团队、扩大本科生和研究生的跨学科研究机会以及参与创业活动的机会，学生的技能将得到提高。拟议的教育课程和外联活动预计将提高公众对应用STEM学科的认识，以满足当前的国家卫生需求。该项目的目标是定量描述决定神经元突触连接强度的蛋白质信号复合物的时空动态，这是由两个谷氨酸门控离子通道（AMPAR受体和NMDA受体）介导的。该研究的目的在于阐明蛋白质信号复合物的形成和调控机制是确定学习和记忆障碍药物干预目标的重要一步。研究计划围绕3个目标组织：1)通过文献挖掘，实验测量大量相互作用蛋白的蛋白质-蛋白质相互作用动力学，开发一种快速测量蛋白质结合动力学的新方法；2)构建NMDAR和AMPAR调控的计算模型，量化各种机制对NMDA和AMPAR定位的影响，识别需要进一步表征的参数；3)利用标记方法结合标准免疫标记技术定量测量蛋白质复合物的形成，利用单分子定位超分辨率显微镜对蛋白质复合物进行成像。教育计划围绕两个目标组织：1)创建一套基于探究的学习模块，向K-12学生教授交通的基本原理，这些模块将呈现给印第安纳州和俄亥俄州农村学校以及城市普渡理工高中的学生；2)通过为本科生和研究生设计和实施新的课程，为当前和未来的普渡大学学生在生物分子检测、生物技术和技术开发领域创造一个多学科的教育途径，这些课程侧重于生物分子工程和运输现象，特别强调工程、化学、生物学和发展扩大的研究和创业活动，提供经验学习的机会，补充技能的获得。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Though more than 10 million people in the USA suffer from learning and memory disorders, the underlying causes of many of these disorders are still not clear. Most of these disorders arise due to disruptions in the regulation of protein networks (called protein signaling complexes) that control the strength of the connections (called synapses) that allow neurons to communicate with each other. Using sophisticated computer modeling and super-resolution microscopes, this project seeks to produce significant new molecular-level understanding of the dynamic organization, in both time and space, of these critical protein complexes. This understanding could lead to the design of new treatments for learning and memory disorders. Furthermore, the research studies planned will provide current and future biomedical engineering students with the multidisciplinary training required to enter careers in the rapidly growing fields of biotechnology and biomedical engineering. Student skills will be enhanced by participation in cross-disciplinary engineering design teams, expanded interdisciplinary research opportunities for undergraduates and graduate students, and opportunities to engage in entrepreneurial activities. The proposed educational curriculum and outreach activities are expected to increase public awareness of the application of STEM disciplines to meet current national health needs.The project's objective is to quantitatively describe the spatiotemporal dynamics of protein signaling complexes that determine the strength of neuronal synaptic connections, which are mediated by two glutamate-gated ion channels (AMPAR receptors and NMDA receptors). The objective is driven by the rationale that elucidating the mechanisms of protein signaling complex formation and regulation is an important step toward identifying targets for pharmaceutical intervention in learning and memory disorders. The Research Plan is organized around 3 aims: 1) Experimentally measure protein-protein interaction dynamics of a large number of interacting proteins through literature mining and developing a new method to rapidly measure protein binding kinetics; 2) Construct computational models of NMDAR and AMPAR regulation to quantify the impact of various mechanisms on NMDA and AMPAR localization and identify parameters that need to be further characterized; 3) Quantitatively measure protein complex formation using labeling methods in conjunction with standard immune-labeling techniques to image protein complexes with single molecule localization super-resolution microscopy. The Educational Plan is organized around 2 aims: 1) Create a set of inquiry-based learning modules to teach fundamental principles of transport to K-12 students that will be presented to students in rural schools in Indiana and Ohio and the urban Purdue Polytechnic High School; 2) Create a multidisciplinary educational pathway for current and future Purdue students to obtain careers in fields of biomolecular detection, biotechnology, and technology development by designing and implementing new courses for undergraduate and graduate students that focus on biomolecular engineering and transport phenomena with special emphasis on the intersection of engineering, chemistry, and biology and developing expanded research and entrepreneurship activities that provide experiential learning opportunities that supplement skill attainment.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.