Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds

Research output: Working paper › Preprint › Research

Standard

Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds. / Ramirez, Santseharay; Fernandez-Antunez, Carlota; Pham, Long V.; Ryberg, Line A.; Feng, Shan; Pedersen, Martin S.; Mikkelsen, Lotte S.; Belouzard, Sandrine; Dubuisson, Jean; Gottwein, Judith M.; Fahnøe, Ulrik; Bukh, Jens.

bioRxiv, 2020.

Research output: Working paper › Preprint › Research

Harvard

Ramirez, S, Fernandez-Antunez, C, Pham, LV, Ryberg, LA, Feng, S, Pedersen, MS, Mikkelsen, LS, Belouzard, S, Dubuisson, J, Gottwein, JM, Fahnøe, U & Bukh, J 2020 'Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds' bioRxiv. https://doi.org/10.1101/2020.10.04.325316

APA

Ramirez, S., Fernandez-Antunez, C., Pham, L. V., Ryberg, L. A., Feng, S., Pedersen, M. S., Mikkelsen, L. S., Belouzard, S., Dubuisson, J., Gottwein, J. M., Fahnøe, U., & Bukh, J. (2020). Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds. bioRxiv. https://doi.org/10.1101/2020.10.04.325316

Vancouver

Ramirez S, Fernandez-Antunez C, Pham LV, Ryberg LA, Feng S, Pedersen MS et al. Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds. bioRxiv. 2020. https://doi.org/10.1101/2020.10.04.325316

Author

Ramirez, Santseharay ; Fernandez-Antunez, Carlota ; Pham, Long V. ; Ryberg, Line A. ; Feng, Shan ; Pedersen, Martin S. ; Mikkelsen, Lotte S. ; Belouzard, Sandrine ; Dubuisson, Jean ; Gottwein, Judith M. ; Fahnøe, Ulrik ; Bukh, Jens. / Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds. bioRxiv, 2020.

Bibtex

@techreport{433d23e533034ce1ace7d7243c3f0f5f,
title = "Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds",
abstract = "Efforts to mitigate COVID-19 include screening of existing antiviral molecules that could be re-purposed to treat SARS-CoV-2 infections. Although SARS-CoV-2 propagates efficiently in African green monkey kidney (Vero) cells, antivirals such as nucleos(t)ide analogs (nucs) often exhibit decreased activity in these cells due to inefficient metabolization. Limited SARS-CoV-2 replication and propagation occurs in human cells, which are the most relevant testing platforms. By performing serial passages of a SARS-CoV-2 isolate in the human hepatoma cell line clone Huh7.5, we selected viral populations with improved viability in human cells. Culture adaptation led to the emergence of a significant number of high frequency changes (>90% of the viral population) in the region coding for the spike glycoprotein, including a deletion of nine amino acids in the N-terminal domain and 3 amino acid changes (E484D, P812R, and Q954H). We demonstrated that the Huh7.5-adapted virus exhibited a >3-Log10 increase in infectivity titers (TCID50) in Huh7.5 cells, with titers of ~8 Log10TCID50/mL, and >2-Log10 increase in the human lung cancer cell line Calu-1, with titers of ~6 Log10TCID50/mL. Culture adaptation in Huh7.5 cells further permitted efficient infection of the otherwise SARS-CoV-2 refractory human lung cancer cell line A549, with titers of ~6 Log10TCID50/mL. The enhanced ability of the virus to replicate and propagate in human cells permitted screening of a panel of nine nucs, including broad-spectrum compounds. Remdesivir, EIDD-2801 and to a limited extent galidesivir showed antiviral effect across these human cell lines, whereas sofosbuvir, uprifosbuvir, valopicitabine, mericitabine, ribavirin, and favipiravir had no apparent activity.",
author = "Santseharay Ramirez and Carlota Fernandez-Antunez and Pham, {Long V.} and Ryberg, {Line A.} and Shan Feng and Pedersen, {Martin S.} and Mikkelsen, {Lotte S.} and Sandrine Belouzard and Jean Dubuisson and Gottwein, {Judith M.} and Ulrik Fahn{\o}e and Jens Bukh",
year = "2020",
doi = "10.1101/2020.10.04.325316",
language = "English",
publisher = "bioRxiv",
type = "WorkingPaper",
institution = "bioRxiv",

}

RIS

TY - UNPB

T1 - Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds

AU - Ramirez, Santseharay

AU - Fernandez-Antunez, Carlota

AU - Pham, Long V.

AU - Ryberg, Line A.

AU - Feng, Shan

AU - Pedersen, Martin S.

AU - Mikkelsen, Lotte S.

AU - Belouzard, Sandrine

AU - Dubuisson, Jean

AU - Gottwein, Judith M.

AU - Fahnøe, Ulrik

AU - Bukh, Jens

PY - 2020

Y1 - 2020

N2 - Efforts to mitigate COVID-19 include screening of existing antiviral molecules that could be re-purposed to treat SARS-CoV-2 infections. Although SARS-CoV-2 propagates efficiently in African green monkey kidney (Vero) cells, antivirals such as nucleos(t)ide analogs (nucs) often exhibit decreased activity in these cells due to inefficient metabolization. Limited SARS-CoV-2 replication and propagation occurs in human cells, which are the most relevant testing platforms. By performing serial passages of a SARS-CoV-2 isolate in the human hepatoma cell line clone Huh7.5, we selected viral populations with improved viability in human cells. Culture adaptation led to the emergence of a significant number of high frequency changes (>90% of the viral population) in the region coding for the spike glycoprotein, including a deletion of nine amino acids in the N-terminal domain and 3 amino acid changes (E484D, P812R, and Q954H). We demonstrated that the Huh7.5-adapted virus exhibited a >3-Log10 increase in infectivity titers (TCID50) in Huh7.5 cells, with titers of ~8 Log10TCID50/mL, and >2-Log10 increase in the human lung cancer cell line Calu-1, with titers of ~6 Log10TCID50/mL. Culture adaptation in Huh7.5 cells further permitted efficient infection of the otherwise SARS-CoV-2 refractory human lung cancer cell line A549, with titers of ~6 Log10TCID50/mL. The enhanced ability of the virus to replicate and propagate in human cells permitted screening of a panel of nine nucs, including broad-spectrum compounds. Remdesivir, EIDD-2801 and to a limited extent galidesivir showed antiviral effect across these human cell lines, whereas sofosbuvir, uprifosbuvir, valopicitabine, mericitabine, ribavirin, and favipiravir had no apparent activity.

AB - Efforts to mitigate COVID-19 include screening of existing antiviral molecules that could be re-purposed to treat SARS-CoV-2 infections. Although SARS-CoV-2 propagates efficiently in African green monkey kidney (Vero) cells, antivirals such as nucleos(t)ide analogs (nucs) often exhibit decreased activity in these cells due to inefficient metabolization. Limited SARS-CoV-2 replication and propagation occurs in human cells, which are the most relevant testing platforms. By performing serial passages of a SARS-CoV-2 isolate in the human hepatoma cell line clone Huh7.5, we selected viral populations with improved viability in human cells. Culture adaptation led to the emergence of a significant number of high frequency changes (>90% of the viral population) in the region coding for the spike glycoprotein, including a deletion of nine amino acids in the N-terminal domain and 3 amino acid changes (E484D, P812R, and Q954H). We demonstrated that the Huh7.5-adapted virus exhibited a >3-Log10 increase in infectivity titers (TCID50) in Huh7.5 cells, with titers of ~8 Log10TCID50/mL, and >2-Log10 increase in the human lung cancer cell line Calu-1, with titers of ~6 Log10TCID50/mL. Culture adaptation in Huh7.5 cells further permitted efficient infection of the otherwise SARS-CoV-2 refractory human lung cancer cell line A549, with titers of ~6 Log10TCID50/mL. The enhanced ability of the virus to replicate and propagate in human cells permitted screening of a panel of nine nucs, including broad-spectrum compounds. Remdesivir, EIDD-2801 and to a limited extent galidesivir showed antiviral effect across these human cell lines, whereas sofosbuvir, uprifosbuvir, valopicitabine, mericitabine, ribavirin, and favipiravir had no apparent activity.

U2 - 10.1101/2020.10.04.325316

DO - 10.1101/2020.10.04.325316

M3 - Preprint

BT - Efficient culture of SARS-CoV-2 in human hepatoma cells enhances viability of the virus in human lung cancer cell lines permitting the screening of antiviral compounds

PB - bioRxiv

ER -

ID: 327472238