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

合作研究：RAPID：定义肺部挥发性化合物特征的分子基础

• 批准号: 2031762
• 负责人: Abhinav Bhushan
• 金额: $ 12.49万
• 依托单位: Illinois Institute of Technology
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2031762&HistoricalAwards=false
• 关键词: Collaborative; Research; RAPID; Molecular; underpinnings

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.

该项目的主要基础研究目标是确定由感染支气管上皮细胞释放的挥发性有机化合物（VOC）的分子基础，并使用分析装置测量它们。目前尚不了解在病毒感染肺细胞中生成VOC信号的生物学机制的基本知识，并且缺乏工程工具和仪器，可以捕获和分析VOC以提供准确的分析信息。这个基础研究项目的预期结果可能会为快速测试的设计提供转化指南，这些测试可以检测美国人群中的SARS-COV-2感染。目前，使用鼻咽拭子的反向转录聚合酶链反应分析证实了Covid-19。鉴于当前的公共卫生紧急情况以及需要防止这种高度传染性病毒的进一步传播的需要，需要采用高度信心，可以动员大量的人，并立即确定需要确认性测试的人。一种有前途的方法是使用为筛查目的而响应感染而形成的挥发性有机化合物的模式。这种技术在病毒感染的快速无创诊断中是无价的。整合细胞生物学，生物医学工程和分析设备的项目中，多学科方法将增强对调节肺VOC的细胞机制的理解，并可能成为非侵入性诊断病毒感染的基础。这个跨学科项目还将在K-12，本科和研究生的生物学与工程交集中提供出色的教育和培训机会。首先，重要的细胞信号通路对VOC的合成的作用。这些途径将在细胞中扰动，然后将细胞感染SARS-COV-2病毒。将确定细胞反应，例如细胞因子释放和转录本的变化。其次，这些细胞将掺入微流体肺芯片装置中。这将有两个目的。第一，器官芯片将为细胞提供生理微环境，以改善其功能反应。二，微流体设备的少量将允许预集中和有效收集VOC。将使用（1）高分辨率气相色谱仪和（2）电子鼻传感器对VOC进行分析。气相色谱仪将通过高分辨率双柱设置进行设置，并带有正交柱涂层，可为VOC提供全面的识别。同时，将使用一个E-Nose传感器来测量VOC，该E-Nose传感器包括纳米复合材料传感器，该传感器根据VOC的吸附改变电阻率。机器学习将在VOC签名上使用，以确定非感染VOC签名的感染性。该平台将发现新科学以调节代谢反应，这将推动生物学和高级仪器发展的基本知识。该快速奖是由蜂窝动力学和功能计划在分子和细胞生物科学划分的划分中颁发的，使用冠状病毒援助，救济和经济安全（CARES）ACT的资金。该奖项反映了NSF的法定任务，并通过使用该基金会的知识分子功能和广泛的影响来评估NSF的法定任务。