Collaborative Research:RAPID:Molecular underpinnings that define volatile compound signature of the lung

该项目的主要基础研究目标是确定受感染的支气管上皮细胞释放的挥发性有机化合物(VOCs)的分子基础，并使用分析设备进行测量。目前尚不清楚在病毒感染的肺细胞中产生VOC信号的生物学机制的基本知识，也缺乏能够捕获和分析VOCs以提供准确分析信息的工程工具和仪器。这一基础研究项目的预期结果可能会为设计能够检测美国人口中SARS-CoV-2感染的快速检测提供翻译指导。目前，新冠肺炎是通过鼻咽拭子的逆转录-聚合酶链式反应分析确认的。鉴于当前的公共卫生紧急情况和防止这种高传染性病毒进一步传播的需要，需要具有高度可信度的护理点筛查方法，这些方法可以动员起来对大量人群进行筛查，并能够立即识别需要进行确证检测的人。一种有希望的方法是利用人体内对感染做出反应而形成的挥发性有机化合物的模式进行筛查。这种技术在病毒感染的快速、非侵入性诊断中将是无价的。在这个整合了细胞生物学、生物医学工程和分析设备的项目中，多学科的方法将增强对调节肺VOCs的细胞机制的理解，并可能成为病毒感染的非侵入性诊断的基础。这个跨学科的项目还将为K-12、本科生和研究生提供生物和工程交叉的优秀教育和培训机会。首先，我们将研究重要的细胞信号通路在VOCs合成中的作用。这些途径将在细胞中受到干扰，之后细胞将感染SARS-CoV-2病毒。将确定细胞反应，如细胞因子的释放和转录本的变化。其次，这些细胞将被整合到微流控芯片肺设备中。这将有两个目的。首先，器官芯片将为细胞提供一个生理微环境，这将改善它们的功能反应。第二，微流控装置的小体积将允许对VOCs进行预浓缩和高效收集。挥发性有机化合物将使用(1)高分辨率气相色谱仪和(2)电子鼻传感器进行分析。该气相色谱仪将配备高分辨率双柱装置和正交柱涂层，这将提供对VOCs的全面鉴定。同时，将使用电子鼻传感器测量VOCs，该传感器由纳米复合材料传感器组成，可以根据VOCs的吸附改变电阻率。机器学习将用于VOC签名，以确定感染性VOC签名与非传染性VOC签名。这一平台将发现代谢反应调节的新科学，这将推动生物学基础知识和先进仪器的发展。这个快速奖项是由分子和细胞生物科学部门的细胞动力学和功能计划利用冠状病毒援助、救济和经济安全(CARE)法案的资金颁发的。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。

The main basic research objectives of this project are to determine the molecular basis for the volatile organic compounds (VOCs) that are released by infected bronchial epithelial cells and to measure them using analytical devices. The fundamental knowledge of the biological mechanisms that generate VOC signals in viral infected lung cells is not currently understood and there is a lack of engineering tools and instrumentation that can capture and analyze the VOCs to provide accurate analytical information. Expected results from this basic research project might provide translational guidance for design of rapid tests that can detect SARS-CoV-2 infection in the US population. Currently, COVID-19 is confirmed using reverse-transcription polymerase chain reaction analysis of nasopharyngeal swabs. Given the current public health emergency and the need to prevent further spread of this highly contagious virus, point of care screening methods are needed that have a high level of confidence, can be mobilized to screen large numbers of people, and can immediately identify persons who require confirmatory testing. One promising approach is to use the pattern of volatile organic compounds that are formed in the body in response to infection for screening purposes. Such a technology would be invaluable in rapid, non-invasive diagnosis of viral infections. The multidisciplinary approach in this project of integrating cell biology, biomedical engineering, and analytical devices will enhance understanding of the cellular mechanisms that regulate lung VOCs and may become the foundation for non-invasive diagnosis of viral infections. This interdisciplinary project will also provide an outstanding educational and training opportunity at the intersection of biology and engineering for K-12, undergraduate, and graduate students.First, the role of important cell signaling pathways on the synthesis of VOCs will be examined. These pathways will be perturbed in the cells after which the cells will be infected with the SARS-CoV-2 virus. The cellular response such as cytokine release and change in transcripts will be determined. Second, the cells will be incorporated in a microfluidic lung-on-a-chip device. This will serve two purposes. One, the organ-chip will provide the cells with a physiological microenvironment which will improve their functional response. Two, the small volume of the microfluidic device will allow pre-concentration and efficient collection of the VOCs. The VOCs will be analyzed using (1) a high-resolution gas chromatograph instrument and (2) an e-nose sensor. The gas chromatograph will be setup with high-resolution dual-column setup with orthogonal column coatings which will provide a comprehensive identification of the VOCs. In parallel, the VOCs will be measured using an e-nose sensor that comprises nanocomposite sensors which change resistivity based on adsorption of VOCs. Machine learning will be used on the VOC signatures to determine an infectious from a non-infectious VOC signature. This platform will uncover new science for regulation of metabolic response which will drive fundamental knowledge of biology and development of advanced instrumentation. This RAPID award is made by the Cellular Dynamics and Function Program in the Division of Molecular and Cellular Biosciences, using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.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.