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

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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.