Kidney Microphysiological Analysis Platforms (MAP) to Explore SARS-CoV-2 Receptors and Inhibitors. A supplement to Parent Grant: Kidney Microphysiological Analysis Platforms (MAP) to Optimize Function

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Abstract Coronavirus disease 2019 (COVID-19) has reached pandemic proportions. Pulmonary and kidney disease are highly prevalent serious consequences of infection with SARS-CoV-2. Kidney Injury Molecule-1 (KIM-1) was identified by Drs Bonventre and Ichimura as the most upregulated protein in the injured kidney proximal tubule. KIM-1, also called TIM-1, is a receptor for hepatitis A, Ebola, Dengue and possibly SARS-CoV1 viruses. We hypothesize that KIM-1 is a receptor for SARS-CoV-2 both in renal tubule epithelial cells and in airway epithelial cells and that JB1, our newly discovered small molecule inhibitor of KIM-1, and/or nanodisc- incorporated KIM-1 ectodomain can be prophylactic and therapeutic agents for COVID-19. We also hypothesize that we can use the high-affinity binding of KIM-1 and ACE2 to the virus to create novel diagnostic devices for the virus. In Specific Aim 1 we will characterize the role of KIM-1 in promoting SARS-CoV-2 entry into kidney and lung epithelia using kidney microphysiological analysis platforms (MAPs) on chip and develop an ultrasensitive chip for the high-throughput evaluation of potential SARS-CoV-2 binding inhibitors. KIM-1 and ACE2 mediated endocytosis of SARS-CoV-2 biomimetic viruses (virosomes) will be compared in kidney and lung epithelial cells. We will evaluate the effects of KIM-1-mediated spike proteins or biomimetic virus cellular adhesion and uptake on production of paracrine factors which activate endothelial cells using a kidney-lung MAP. In order to understand binding and/or uptake kinetics of SARS-CoV-2 and characterize potential inhibitors we will develop an ultrasensitive nanoplasmonic triplets-based rapid lateral flow diagnostic chip for a rapid and sensitive inhibition assay using the kidney-lung MAP. This approach can also be used for point of care diagnostic testing for the virus. In Specific Aim 2 we will evaluate the efficacy of JB1, soluble KIM-1 ectodomain and nanodisc-incorporated KIM-1 or ACE2 to inhibit binding and internalization of SARS-CoV-2 biomimetic viruses by kidney and lung cells using an integrated lung-kidney MAP on chip. Potential inhibitors will be tested to evaluate whether they compete with S-protein and/or biomimetic virus binding and reduce IL-6 production. Binding affinity and kinetics between KIM-1 variants or ACE2 either as free ectodomains or incorporated into nanodiscs and the Spike protein will be measured using MicroScale Thermophoresis (MST) and Biolayer Interferometry.