Evaluation of SARS-CoV-2 entry, inflammation and new therapeutics in human lung tissue cells

The development of physiological models that reproduce SARS-CoV-2 infection in primary human cells will be instrumental to identify host-pathogen interactions and potential therapeutics. Here, using cell suspensions directly from primary human lung tissues (HLT), we have developed a rapid platform for the identification of viral targets and the expression of viral entry factors, as well as for the screening of viral entry inhibitors and anti-inflammatory compounds. The direct use of HLT cells, without long-term cell culture and in vitro differentiation approaches, preserves main immune and structural cell populations, including the most susceptible cell targets for SARS-CoV-2; alveolar type II (AT-II) cells, while maintaining the expression of proteins involved in viral infection, such as ACE2, TMPRSS2, CD147 and AXL. Further, antiviral testing of 39 drug candidates reveals a highly reproducible method, suitable for different SARS-CoV-2 variants, and provides the identification of new compounds missed by conventional systems, such as VeroE6. Using this method, we also show that interferons do not modulate ACE2 expression, and that stimulation of local inflammatory responses can be modulated by different compounds with antiviral activity. Overall, we present a relevant and rapid method for the study of SARS-CoV-2.
This work was primarily supported by a grant from the Health Department of the Government of Catalonia (DGRIS 1_5) to M.G and MJ.B. This work was additionally supported in part by the Spanish Health Institute Carlos III (ISCIII, PI17/01470 to M.G; PI19CIII/00004 to J.A; PI21CIII/00025 to J.G-P and COV20-00679 (MPY 222-20) to J.G-P), the Spanish Secretariat of Science and Innovation and FEDER funds (grant RTI2018-101082-B-I00 [MINECO/FEDER]) to MJ.B, the Spanish AIDS network Red Temática Cooperativa de Investigación en SIDA and the European Regional Development Fund (ERDF) (RD16/0025/0007 to MJ.B and RD16CIII/0002/0001 to J.A), the Fundació La Marató TV3 (grants 201805-10FMTV3 and 202104FMTV3 to MJ.B; 201814-10FMTV3 and 202112FMTV3 to M.G), the Gilead fellowships GLD19/00084 to M.G and GLD18/00008 to MJ.B and the Becas Taller Argal 2020 to MJ.B. Salary for MJ.B is supported by the Miguel Servet program funded by the Spanish Health Institute Carlos III (CP17/00179). N.M and D.P are supported by a Ph.D. fellowship from the Vall d’Hebron Institut de Recerca (VHIR).The funders had no role in study design, data collection and analysis, the decision to publish, or preparation of the manuscript.