TY - JOUR
T1 - RK-33, a small molecule inhibitor of host RNA helicase DDX3, suppresses multiple variants of SARS-CoV-2
AU - Vesuna, Farhad
AU - Akhrymuk, Ivan
AU - Smith, Amy
AU - Winnard, Paul T.
AU - Lin, Shih Chao
AU - Panny, Lauren
AU - Scharpf, Robert
AU - Kehn-Hall, Kylene
AU - Raman, Venu
N1 - Funding Information:
We wish to acknowledge Robert O’Malley and Bob Cole at the Johns Hopkins University School of Medicine Mass Spectrometry and Proteomics Core for performing the proteomics experiments and Co-Diagnostics, Inc. (Salt Lake City, UT, USA) for providing Logix Smart COVID-19 real-time PCR reagents.
Funding Information:
This work was supported by the NIH (Grant R01CA207208) and FAMRI for funding to VR.
Publisher Copyright:
Copyright © 2022 Vesuna, Akhrymuk, Smith, Winnard, Lin, Panny, Scharpf, Kehn-Hall and Raman.
PY - 2022/8/25
Y1 - 2022/8/25
N2 - SARS-CoV-2, the virus behind the deadly COVID-19 pandemic, continues to spread globally even as vaccine strategies are proving effective in preventing hospitalizations and deaths. However, evolving variants of the virus appear to be more transmissive and vaccine efficacy toward them is waning. As a result, SARS-CoV-2 will continue to have a deadly impact on public health into the foreseeable future. One strategy to bypass the continuing problem of newer variants is to target host proteins required for viral replication. We have used this host-targeted antiviral (HTA) strategy that targets DDX3X (DDX3), a host DEAD-box RNA helicase that is usurped by SARS-CoV-2 for virus production. We demonstrated that targeting DDX3 with RK-33, a small molecule inhibitor, reduced the viral load in four isolates of SARS-CoV-2 (Lineage A, and Lineage B Alpha, Beta, and Delta variants) by one to three log orders in Calu-3 cells. Furthermore, proteomics and RNA-seq analyses indicated that most SARS-CoV-2 genes were downregulated by RK-33 treatment. Also, we show that the use of RK-33 decreases TMPRSS2 expression, which may be due to DDX3s ability to unwind G-quadraplex structures present in the TMPRSS2 promoter. The data presented support the use of RK-33 as an HTA strategy to control SARS-CoV-2 infection, irrespective of its mutational status, in humans.
AB - SARS-CoV-2, the virus behind the deadly COVID-19 pandemic, continues to spread globally even as vaccine strategies are proving effective in preventing hospitalizations and deaths. However, evolving variants of the virus appear to be more transmissive and vaccine efficacy toward them is waning. As a result, SARS-CoV-2 will continue to have a deadly impact on public health into the foreseeable future. One strategy to bypass the continuing problem of newer variants is to target host proteins required for viral replication. We have used this host-targeted antiviral (HTA) strategy that targets DDX3X (DDX3), a host DEAD-box RNA helicase that is usurped by SARS-CoV-2 for virus production. We demonstrated that targeting DDX3 with RK-33, a small molecule inhibitor, reduced the viral load in four isolates of SARS-CoV-2 (Lineage A, and Lineage B Alpha, Beta, and Delta variants) by one to three log orders in Calu-3 cells. Furthermore, proteomics and RNA-seq analyses indicated that most SARS-CoV-2 genes were downregulated by RK-33 treatment. Also, we show that the use of RK-33 decreases TMPRSS2 expression, which may be due to DDX3s ability to unwind G-quadraplex structures present in the TMPRSS2 promoter. The data presented support the use of RK-33 as an HTA strategy to control SARS-CoV-2 infection, irrespective of its mutational status, in humans.
KW - DDX3 inhibitor
KW - RK-33
KW - RNA helicase
KW - SARS-CoV-2
KW - TMPRSS2
KW - host-targeted antiviral
KW - viral isolates
UR - http://www.scopus.com/inward/record.url?scp=85138011649&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85138011649&partnerID=8YFLogxK
U2 - 10.3389/fmicb.2022.959577
DO - 10.3389/fmicb.2022.959577
M3 - Article
C2 - 36090095
AN - SCOPUS:85138011649
SN - 1664-302X
VL - 13
JO - Frontiers in Microbiology
JF - Frontiers in Microbiology
M1 - 959577
ER -