CAREER: Extracellular Hydrogen Peroxide and Nitric Oxide Detection and Quantification Via Biocompatible Carbon Nanotubes

活性氧和氮物种(ROS和RNS)是细胞信号传递的重要组成部分，已知它们在健康和疾病/炎症中发挥重要作用--既有有益的，也有有害的；然而，它们在健康和疾病状态下的浓度和机制在很大程度上是未知的。这个职业项目将通过使用纳米传感器来量化ROS/RNS浓度来促进对ROS和RNS水平如何影响细胞和组织功能的理解。最终，了解ROS/RNS水平与疾病细胞的关系，并使用新的传感器随后量化这些浓度，可以快速检测受感染的个人，从而改善诊断和健康结果。与这项研究相结合的是旨在向奥舍终身学习研究所(OLLI)的农村中学生、本科生和研究生以及50岁以上的成年人传授纳米技术和传感器的教育努力。大学生将在直接为研究目标做出贡献的同时接受培训、指导和实践经验，并将通过协助外展计划提高他们的科学交流技能，在教育和研究目标之间建立互惠关系。中学和OLLI项目的参与者将增强他们对纳米技术和传感器的理解，并能够与他人分享他们新学到的知识，将该项目的影响扩大到直接参与该项目的个人。研究人员的长期职业目标是开发和使用单壁碳纳米管(SWNT)传感器来了解生物现象，同时为下一代在纳米技术和传感器开发方面的职业生涯做好准备。为了实现这一目标，本项目致力于通过开发同时定量测定过氧化氢(H_2O_2)和一氧化氮(NO)的单壁碳纳米管传感器系统，促进对细胞信号转导中ROS和RNS的理解。研究目标包括：(1)开发一种可同时定量测定过氧化氢和一氧化氮浓度的无菌平台系统；(2)确定与乳腺、皮肤和淋巴细胞相关的细胞外过氧化氢和一氧化氮浓度分布；以及(3)评估过氧化氢和一氧化氮在细胞增殖、炎症标志物释放和细胞迁移中的重要性。假设病变细胞的过氧化氢和一氧化氮水平明显较高，增殖率、迁移率和炎症因子可以随着活性物质的增加(导致增加)或清除(导致减少)而增加或减少。这个程序代表了一种新的和变革性的方法来定量H_2O_2和NO，并将阐明以前无法获得的关于细胞信号转导的基本事实。此外，这项研究代表了碳纳米管领域的一个新里程碑，该领域在很大程度上专注于开发新的传感器，而不是使用这些独特的传感器来了解生物系统的功能。随着更多传感器的开发，这项研究的结果将成为未来检测其他小分子的路线图。该项目由生物传感计划和既定的刺激竞争研究计划(EPSCoR)联合资助。该奖项反映了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。

Reactive oxygen and nitrogen species (ROS and RNS) are essential components in cell signaling and they are known to play an important role in health and disease/inflammation—both helpful and harmful; however, their concentrations and mechanisms during healthy and diseased states are largely unknown. This CAREER project will advance the understanding of how ROS and RNS levels impact the function of cells and tissues using nano-scale sensors to quantify ROS/RNS concentrations. Ultimately, understanding how ROS/RNS levels correlate with diseased cells and using novel sensors to subsequently quantify these concentrations could provide quick detection of infected individuals, thus leading to improved diagnostics and health outcomes. Integrated with this research are educational efforts that aim to teach rural middle school students, undergraduate and graduate students, and adults over 50 at the Osher Lifelong Learning Institute (OLLI) about nanotechnology and sensors. University students will receive training, mentorship, and hands-on experience while directly contributing to the research objectives and will enhance their science communication skills by assisting with the outreach programs, creating a reciprocal relationship between the education and research objectives. Participants in the middle school and OLLI programs will enhance their understanding of nanotechnology and sensors and be able to share their newly acquired knowledge with others, extending the project’s impact beyond the individuals directly participating in the programs.The investigator’s long-term career goal is to develop and use single walled carbon nanotube (SWNT) sensors to learn about biological phenomena while preparing the next generation for careers in nanotechnology and sensor development. Towards this goal, this project focuses on advancing the understanding of ROS and RNS in cell signaling by developing a single walled carbon nanotube sensor system that quantifies hydrogen peroxide (H2O2) and nitric oxide (NO) simultaneously. Research objectives include to: (1) develop a sterile platform system that can simultaneously quantify H2O2 and NO concentrations; (2) identify the concentration profiles of extracellular H2O2 and NO associated with breast, skin, and lymphocyte cells; and (3) evaluate the importance of H2O2 and NO in cellular proliferation, release of inflammatory markers, and cell migration. It is hypothesized that diseased cells will have significantly higher levels of H2O2 and NO and that proliferation rate, migration, and inflammatory factors can be increased or decreased with the addition (causing an increase) or scavenging (causing a decrease) of reactive species. This program represents a novel and transformative approach for quantifying H2O2 and NO and will illuminate basic facts about cell signaling that were previously unattainable. Furthermore, this research represents a new milestone in the field of carbon nanotubes, which has largely focused on developing new sensors rather than using these unique sensors to understand how biological systems function. Findings from this research will serve as a roadmap for detecting other small molecules in the future as additional sensors are developed.This project is jointly funded by the Biosensing Program and the Established Program to Stimulate Competitive Research (EPSCoR).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.