Identification of druggable host dependency factors shared by multiple SARS-CoV-2 variants of concern

Abstract: Identification of druggable host dependency factors shared by multiple SARS-CoV-2 variants of concern A N U SC RI PT 1 Department of Molecular Medicine, University of Padua, Padua 35121, Italy 2 Department of Biology, Armenise/Harvard Pluripotent Stem Cell Biology Laboratory, University of Padua, Padua 35131, Italy 3 Department. of Surgery, Oncology and Gastroenterology, University of Padua, Padua 35128, Italy 4 Veneto Institute of Oncology IOV-IRCCS, Padua 35128, Italy 5 Department of Comparative Biomedicine and Food Science, University of Padua, Padua 35020, Italy 6 Telethon Institute of Genetics and Medicine (TIGEM), Armenise/Harvard Laboratory of Integrative Genomics, Pozzuoli 80078, Italy 7 Department of Translational Medicine, Federico II University, Naples 80138, Italy 8 School for Advanced Studies, Genomics and Experimental Medicine Program, University of Naples “Federico II”, Naples 80138, Italy 9 Microbiology and Virology Unit, Padua University Hospital, Padua 35128, Italy † O RI G IN A L U N ED IT ED M These authors contributed equally to this work. * Correspondence to: Sara N. Richter, E-mail: sara.richter@unipd.it; Marco Montagner, Email: marco.montagner@unipd.it; Graziano Martello, E-mail: graziano.martello@unipd.it © The Author(s) 2024. Published by Oxford University Press on behalf of Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Ilaria Frasson1,†, Linda Diamante1,2,†, Manuela Zangrossi1,†, Elena Carbognin2, Anna Dalla Pietà3, Alessandro Penna3, Antonio Rosato3,4, Ranieri Verin5, Filippo Torrigiani5, Cristiano Salata1, Marìa Paula Dizanzo1, Lorenzo Vaccaro6,7, Davide Cacchiarelli6,7,8, Sara N. Richter1,9,*, Marco Montagner1,*, Graziano Martello2,* Abstract The high mutation rate of SARS-CoV-2 leads to the emergence of multiple variants, some of which are resistant to vaccines and drugs targeting viral elements. Targeting host dependency factors, e.g. cellular proteins required for viral replication, would variants induce conserved cellular responses and exploit the same core host factors. To this end, we compared three variants of concern and found that the host transcriptional response was conserved, differing only in kinetics and magnitude. PT Through CRISPR screening, we identified host genes required for infection by each RI variant. Most of the genes were shared by multiple variants. We validated our hits with small molecules and repurposed Food and Drug Administration-approved drugs. SC All the drugs were highly active against all the variants tested, including new variants that emerged during the study (Delta and Omicron). Mechanistically, we identified U reactive oxygen species production as a key step in early virus replication. N Antioxidants such as N-acetyl cysteine (NAC) were effective against all the variants A in both human lung cells and a humanised mouse model. Our study supports the use M of available antioxidant drugs, such as NAC, as a general and effective..