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。脑外伤治疗发展中的一个主要挑战是对由脑外伤引发的进行性疾病的不完全理解。因此，这项职业奖的目标是设计技术来测量与脑损伤相关的活动，并使用这项技术来了解脑损伤背后的分子机制。对脑损伤相关分子机制的新认识可以指导脑损伤和其他脑部疾病的新疗法的发展。这项研究将为不同的本科生和研究生群体提供以研究为基础的培训机会，为他们进入科学、技术、工程和数学(STEM)大军做准备。这项研究的想法和概念将与高中教师合作纳入教案，以促进在STEM领域的广泛参与。在开发新的治疗方法和临床治疗方面存在的一个挑战是缺乏能够衡量疾病严重程度和预测长期结果的生物标记物。蛋白水解酶是脑损伤疾病进展的关键分子，也是治疗脑损伤的候选药物。因此，测量TBI相关的蛋白水解酶活性对于了解长期生物学结果和更好地了解疾病可能是至关重要的。然而，目前还没有工具来测量颅脑损伤后活的生物体中的蛋白酶活性。这项职业计划的目标是设计技术来测量体内的蛋白酶活性，并研究蛋白酶活性在脑外伤疾病进展和治疗中的作用。作为第二个目标，这项技术也将被开发为一种诊断方法，以补充脑外伤现有的诊断范例。拟议中的“纳米传感器”技术是基于纳米材料能够通过受损的血脑屏障被动积聚到受损的大脑中，并对脑损伤后升高的蛋白酶活性做出反应的能力。将被设计成纳米传感器的设计标准包括检测多种蛋白酶的能力，从生物液中测量的能力，以及主动定位的能力。这项技术将用于研究在不同程度的脑损伤和存在药物抑制的情况下的蛋白酶活性。该纳米传感器还将被设计为与纸基分析相结合，以实现低资源、快速读出，作为临床上现有成像和行为诊断的补充诊断工具。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。由于蛋白水解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，因此测量蛋白水解酶活性的工具和对疾病中蛋白水解酶活性的更深入了解具有广泛应用的潜力。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.