CAREER: An engineered nanosensor to measure in vivo protease activity in traumatic brain injury

创伤性脑损伤（TBI）每年影响数百万美国人。创伤性脑损伤治疗发展的主要挑战是对创伤性脑损伤引发的进行性疾病的不完全了解。因此，CAREER奖的目标是设计测量TBI相关活性的技术，并利用该技术了解TBI的分子机制。对TBI相关分子机制的新认识可以指导TBI和其他脑部疾病的新疗法的发展。该研究将为不同群体的本科生和研究生提供基于研究的培训机会，为他们进入科学技术工程和数学（STEM）劳动力做好准备。研究中的想法和概念将与高中教师合作纳入课程计划，以促进STEM领域的广泛参与。在TBI的新疗法和临床管理发展中存在的一个挑战是缺乏可以衡量疾病严重程度和预测长期结果的生物标志物。蛋白酶是TBI疾病进展的关键分子，是TBI治疗抑制的候选分子。因此，测量脑外伤相关蛋白酶活性可能对了解长期生物学结果和更好地了解疾病至关重要。然而，目前还没有工具来测量脑外伤后活生物体的蛋白酶活性。本CAREER提案的目标是设计技术来测量体内蛋白酶活性，并研究蛋白酶活性在TBI疾病进展和治疗中的作用。作为第二个目标，该技术还将被开发为一种诊断方法，以补充现有的TBI诊断范例。即将开发的“纳米传感器”技术是基于纳米级材料的能力，它可以通过受损的血脑屏障被动地积聚到受伤的大脑中，并对脑外伤后蛋白酶活性升高做出反应。纳米传感器的设计标准包括检测多种蛋白酶的能力，可从生物液体中测量，并具有主动靶向性。该技术将用于研究蛋白酶活性，以应对不同程度的创伤性脑损伤和存在的药物抑制。纳米传感器还将被设计成与基于纸张的检测相结合，以获得低资源、快速的读数，作为一种诊断工具，补充临床现有的成像和行为诊断。这项研究将产生一种多路和靶向的蛋白酶活性纳米传感器。由于蛋白酶在包括传染病在内的多种疾病中发挥着不可或缺的作用，因此测量蛋白酶活性的工具和对蛋白酶在疾病中的活性的更深入的了解具有广泛应用的潜力。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。

Traumatic brain injury (TBI) affects millions of Americans annually. A major challenge in the development of TBI therapies is an incomplete understanding of the progressive disease that is initiated by TBI. Therefore, the goal of this CAREER award is to engineer technology to measure TBI-associated activity and use this technology to understand the molecular mechanisms that underlie TBI. New understanding of TBI-associated molecular mechanisms can guide the development of new therapies for TBI and other brain disorders. The research will provide research-based training opportunities for a diverse group of undergraduate and graduate students to prepare them to enter the science technology engineering and mathematics (STEM) workforce. Ideas and concepts from the research will be incorporated into lesson plans in collaboration with high school teachers in order to promote broad participation in STEM fields. One existing challenge in the development of new therapeutics and clinical management of TBI is a lack of biomarkers that can measure the severity of disease and predict long-term outcomes. Proteases are key molecules in TBI disease progression and are candidates for therapeutic inhibition in TBI. Therefore, the measurement of TBI-associated protease activity may be critical to understand long-term biological outcomes and provide a greater understanding of disease. However, currently, there are no tools to measure protease activity in a living organism after TBI. The goal of this CAREER proposal is to engineer technology to measure protease activity in vivo and study the role of protease activity in TBI disease progression and therapeutic treatment. As a second goal, the technology will also be developed as a diagnostic to complement existing diagnostic paradigms in TBI. The proposed ‘nanosensor’ technology that will be developed is based on the ability of nanometer scaled materials to passively accumulate into the injured brain across the damaged blood-brain barrier and respond to elevated protease activity after TBI. Design criteria that will be engineered into the nanosensor include the ability to detect multiple proteases, be measured from biological fluids, and be actively targeted. The technology will be used to study protease activity in response to varying severities of TBI and in the presence of pharmacologic inhibition. The nanosensor will also be designed to integrate with a paper-based assay for a low-resource, rapid readout as a diagnostic tool that complements existing imaging and behavioral diagnostics in the clinic. This research will yield a multiplexed and targeted nanoscale sensor of protease activity. Due to the integral role proteases have across multiple diseases, including infectious diseases, tools to measure protease activity and a greater understanding of protease activity in disease have the potential to be broadly applicable.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.