tetano
Editor, Senior Moderator
Antiviral Res
. 2024 Aug 22:105992.
doi: 10.1016/j.antiviral.2024.105992. Online ahead of print. Anti-SARS-CoV-2 gapmer antisense oligonucleotides targeting the main protease region of viral RNA
Masako Yamasaki[SUP] 1 [/SUP], Wakana Saso[SUP] 2 [/SUP], Takuya Yamamoto[SUP] 3 [/SUP], Masayoshi Sato[SUP] 3 [/SUP], Hiroko Takagi[SUP] 3 [/SUP], Tetsuya Hasegawa[SUP] 4 [/SUP], Yuji Kozakura[SUP] 5 [/SUP], Hiroyuki Yokoi[SUP] 6 [/SUP], Hirofumi Ohashi[SUP] 7 [/SUP], Kana Tsuchimoto[SUP] 7 [/SUP], Rina Hashimoto[SUP] 8 [/SUP], Shuetsu Fukushi[SUP] 9 [/SUP], Akihiko Uda[SUP] 10 [/SUP], Masamichi Muramatsu[SUP] 11 [/SUP], Kazuo Takayama[SUP] 8 [/SUP], Ken Maeda[SUP] 10 [/SUP], Yoshimasa Takahashi[SUP] 7 [/SUP], Tsuyoshi Nagase[SUP] 3 [/SUP], Koichi Watashi[SUP] 12 [/SUP]
Affiliations
Given the worldwide risk for the outbreak of emerging/re-emerging respiratory viruses, establishment of new antiviral strategies is greatly demanded. In this study, we present a scheme to identify gapmer antisense oligonucleotides (ASOs) targeting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA that efficiently inhibit viral replication. We synthesized approximately 300 gapmer ASOs designed to target various SARS-CoV-2 RNA regions and evaluated their activity in cell-based assays. Through a multistep screening in cell culture systems, we identified that ASO#41, targeting the coding region for viral main protease, reduced SARS-CoV-2 RNA levels in infected cells and inhibited virus-induced cytopathic effects. Antiviral effect of ASO#41 was also observed in iPS cell-derived human lung organoids. ASO#41 depleted intracellular viral RNAs during genome replication in an endogenous RNaseH-dependent manner. ASO#41 showed a wide range of antiviral activity against SARS-CoV-2 variants of concern including Alpha, Delta, and Omicron. Intranasal administration to mice exhibited intracellular accumulation of ASO#41 in the lung and significantly reduced the viral infectious titer, with milder body weight loss due to SARS-CoV-2 infection. Further chemical modification with phosphoryl guanidine-containing backbone linkages provided an elevation of anti-SARS-CoV-2 activity, with 23.4 nM of 50% antiviral inhibitory concentration, one of the strongest anti-SARS-CoV-2 ASOs reported so far. Our study presents an approach to identify active ASOs against SARS-CoV-2, which is potentially useful for establishing an antiviral strategy by targeting genome RNA of respiratory viruses.
Keywords: RNaseH; SARS-CoV-2; antisense; coronavirus; gapmer.
. 2024 Aug 22:105992.
doi: 10.1016/j.antiviral.2024.105992. Online ahead of print. Anti-SARS-CoV-2 gapmer antisense oligonucleotides targeting the main protease region of viral RNA
Masako Yamasaki[SUP] 1 [/SUP], Wakana Saso[SUP] 2 [/SUP], Takuya Yamamoto[SUP] 3 [/SUP], Masayoshi Sato[SUP] 3 [/SUP], Hiroko Takagi[SUP] 3 [/SUP], Tetsuya Hasegawa[SUP] 4 [/SUP], Yuji Kozakura[SUP] 5 [/SUP], Hiroyuki Yokoi[SUP] 6 [/SUP], Hirofumi Ohashi[SUP] 7 [/SUP], Kana Tsuchimoto[SUP] 7 [/SUP], Rina Hashimoto[SUP] 8 [/SUP], Shuetsu Fukushi[SUP] 9 [/SUP], Akihiko Uda[SUP] 10 [/SUP], Masamichi Muramatsu[SUP] 11 [/SUP], Kazuo Takayama[SUP] 8 [/SUP], Ken Maeda[SUP] 10 [/SUP], Yoshimasa Takahashi[SUP] 7 [/SUP], Tsuyoshi Nagase[SUP] 3 [/SUP], Koichi Watashi[SUP] 12 [/SUP]
Affiliations
- PMID: 39181215
- DOI: 10.1016/j.antiviral.2024.105992
Given the worldwide risk for the outbreak of emerging/re-emerging respiratory viruses, establishment of new antiviral strategies is greatly demanded. In this study, we present a scheme to identify gapmer antisense oligonucleotides (ASOs) targeting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA that efficiently inhibit viral replication. We synthesized approximately 300 gapmer ASOs designed to target various SARS-CoV-2 RNA regions and evaluated their activity in cell-based assays. Through a multistep screening in cell culture systems, we identified that ASO#41, targeting the coding region for viral main protease, reduced SARS-CoV-2 RNA levels in infected cells and inhibited virus-induced cytopathic effects. Antiviral effect of ASO#41 was also observed in iPS cell-derived human lung organoids. ASO#41 depleted intracellular viral RNAs during genome replication in an endogenous RNaseH-dependent manner. ASO#41 showed a wide range of antiviral activity against SARS-CoV-2 variants of concern including Alpha, Delta, and Omicron. Intranasal administration to mice exhibited intracellular accumulation of ASO#41 in the lung and significantly reduced the viral infectious titer, with milder body weight loss due to SARS-CoV-2 infection. Further chemical modification with phosphoryl guanidine-containing backbone linkages provided an elevation of anti-SARS-CoV-2 activity, with 23.4 nM of 50% antiviral inhibitory concentration, one of the strongest anti-SARS-CoV-2 ASOs reported so far. Our study presents an approach to identify active ASOs against SARS-CoV-2, which is potentially useful for establishing an antiviral strategy by targeting genome RNA of respiratory viruses.
Keywords: RNaseH; SARS-CoV-2; antisense; coronavirus; gapmer.