Unraveling the antiviral activity of plitidepsin against SARS-CoV-2 by subcellular and morphological analysis
Por:
Sachse, M, Tenorio, R, Castro, IFD, Munoz-Basagoiti, J, Perez-Zsolt, D, Raich-Regu, D, Rodon, J, Losada, A, Avil, P, Cuevas, C, Paredes, R, Segales, J, Clotet, B, Vergara-Alert, J, Izquierdo-Useros, N and Risco, C
Publicada:
1 abr 2022
Ahead of Print:
1 mar 2022
Resumen:
The pandemic caused by the new coronavirus SARS-CoV-2 has made evident the need for broad-spectrum, efficient antiviral treatments to combat emerging and re-emerging viruses. Plitidepsin is an antitumor agent of marine origin that has also shown a potent pre-clinical efficacy against SARS-CoV-2. Plitidepsin targets the host protein eEF1A (eukaryotic translation elongation factor 1 alpha) and affects viral infection at an early, post-entry step. Because electron microscopy is a valuable tool to study virus-cell interactions and the mechanism of action of antiviral drugs, in this work we have used transmission electron microscopy (TEM) to evaluate the effects of plitidepsin in SARS-CoV-2 infection in cultured Vero E6 cells 24 and 48h post-infection. In the absence of plitidepsin, TEM morphological analysis showed double-membrane vesicles (DMVs), organelles that support coronavirus genome replication, single-membrane vesicles with viral particles, large vacuoles with groups of viruses and numerous extracellular virions attached to the plasma membrane. When treated with plitidepsin, no viral structures were found in SARS-CoV-2-infected Vero E6 cells. Immunogold detection of SARS-CoV-2 nucleocapsid (N) protein and double-stranded RNA (dsRNA) provided clear signals in cells infected in the absence of plitidepsin, but complete absence in cells infected and treated with plitidepsin. The present study shows that plitidepsin blocks the biogenesis of viral replication organelles and the morphogenesis of virus progeny. Electron microscopy morphological analysis coupled to immunogold labeling of SARS-CoV-2 products offers a unique approach to understand how antivirals such as plitidepsin work.
Filiaciones:
Sachse, M:
CSIC, Ctr Nacl Biotecnol, 28049 Madrid, Spain
Tenorio, R:
CSIC, Ctr Nacl Biotecnol, 28049 Madrid, Spain
Castro, IFD:
CSIC, Ctr Nacl Biotecnol, 28049 Madrid, Spain
Munoz-Basagoiti, J:
IrsiCaixa AIDS Res Inst, Badalona 08916, Spain
:
IrsiCaixa AIDS Res Inst, Badalona 08916, Spain
Raich-Regu, D:
IrsiCaixa AIDS Res Inst, Badalona 08916, Spain
Rodon, J:
Campus Univ Autonoma Barcelona UAB, Ctr Recerca Sanitat Anim CReSA, Unitat mixta Invest IRTA UAB Sanitat Anim, 08193 Bellaterra, Catalonia, Spain
Avil, P:
PharmaMar SA, Madrid 28770, Spain
Cuevas, C:
PharmaMar SA, Madrid 28770, Spain
:
IrsiCaixa AIDS Res Inst, Badalona 08916, Spain
Segales, J:
Campus Univ Autonoma Barcelona UAB, Ctr Recerca Sanitat Anim CReSA, IRTA, Programa Sanitat Anim, Bellaterra 08193, Catalonia, Spain
Univ Autonoma Barcelona UAB, Dept Sanitat & Anat Anim, Fac Vet, Campus UAB, Bellaterra 08193, Catalonia, Spain
:
IrsiCaixa AIDS Res Inst, Badalona 08916, Spain
Germans Trias i Pujol Res Inst IGTP, Can Ruti Campus, Badalona 08916, Spain
Univ Vic, Cent Univ Catalonia UV UCC, Barcelona 08500, Spain
Vergara-Alert, J:
Campus Univ Autonoma Barcelona UAB, Ctr Recerca Sanitat Anim CReSA, Unitat mixta Invest IRTA UAB Sanitat Anim, 08193 Bellaterra, Catalonia, Spain
:
IrsiCaixa AIDS Res Inst, Badalona 08916, Spain
Germans Trias i Pujol Res Inst IGTP, Can Ruti Campus, Badalona 08916, Spain
Risco, C:
CSIC, Ctr Nacl Biotecnol, 28049 Madrid, Spain
Green Published, Green Submitted, hybrid
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