Lung-on-a-Chip Disease Models for Efficacy Testing (COVID-19 Competitive Revision)

PROJECT SUMMARY This COMPETITIVE REVISION application is being submitted to expand the scope of our ongoing NIHgrant UH3HL141797 in order to leverage our human organ-on-a-chip (Organ Chip) microfluidic culture devicesfor the rapid development and assessment of potential therapeutic agents for COVID-19. Our ongoing UH3grant supports the development of human Lung Chips as in vitro preclinical tools for rapid discovery of newtherapeutics for viral pandemics caused by influenza. In recent studies, we showed that highly differentiatedhuman cells in our Lung Chips, as well as human intestinal cells within Intestine Chips we developed, expresshigh levels of ACE2 and TMPRSS2 that mediate SARS-CoV-2 virus (CoV2) infection. We also were able toinfect these Organ Chips with CoV2 spike protein-expressing viral pseudoparticles (CoV2pp) that closely mimicthe effects of native CoV2 virus when tested against multiple FDA approved drugs in cell-based assays.Human Lung Chips were also shown to be more stringent models for assessing potential COVID19 inhibitoryactivity as only a subset of these drugs significantly inhibited entry of the CoV2pp when administered underflow on-chip at their maximum concentration (Cmax) in human blood reported in clinical studies. Here, wepropose to use human Intestine and Lung Chips in combination with computational discovery and syntheticchemistry approaches to develop broad-spectrum coronavirus therapeutics that would both help infectedCOVID19 patients now, and allow us to be prepared to prevent infections by related pandemic viruses thatemerge in the future. In preliminary studies, multiple novel compounds designed with our computational toolsexhibited significant inhibitory activities when tested against both CoV2pp and native CoV2 virus in cell basedassays. Thus, our Specific Aims include: 1) to use computational and synthetic chemistry approaches tocreate new compounds that are predicted to inhibit infection by CoV2 virus and related coronaviruses, 2) toprioritize active molecules by analyzing their structure-activity relationships in cell-based assays infected withnative CoV2 and related coronaviruses, 3) to identify lead compounds and effective doses based on inhibitionof infection and host inflammatory responses in human Organ Chips using native coronaviruses, and 4) tocarry out pharmacokinetic studies in mice coupled with iterative chemical synthesis and testing in cell-basedassays to optimize the pharmaceutical properties and safety of the lead compounds, while retaining efficacy.Through this effort, we will identify new compounds that demonstrate broad spectrum inhibiting activitiesagainst CoV2 as well as related coronaviruses, and generate pharmacokinetic data necessary to move thesedrugs into animal validation studies and, eventually, human clinical trials. This work will also further establishthe value of human Organ Chips as preclinical tools for accelerating drug development.