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Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses
Published on: April 4, 2019
Profiling Ribonucleotide and Deoxyribonucleotide Pools Perturbed by Remdesivir in Human Bronchial Epithelial Cells
Yan Li1, Hui-Xia Zhang1, Wen-Di Luo1
1State Key Laboratory of Quality Research in Chinese Medicines, Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Guangdong-Hong Kong-Macao Joint Laboratory of Respiratory Infectious Disease (Macau University of Science and Technology), Taipa, Macau, China.
Remdesivir impacts cellular nucleotide pools, inhibiting RNA and DNA synthesis. Exacerbating this imbalance shows promise for enhancing antiviral efficacy against SARS-CoV-2 and treating COVID-19.
Area of Science:
- Biochemistry
- Virology
- Cell Biology
Background:
- Remdesivir (RDV) shows moderate efficacy against SARS-CoV-2, but limited survival rates necessitate improved treatments for COVID-19.
- RDV, a nucleotide analog, affects host cell nucleotide biosynthesis, impacting viral replication.
Purpose of the Study:
- To investigate the effects of RDV on cellular nucleotide pools, RNA transcription, and DNA replication.
- To explore strategies for enhancing RDV's antiviral efficacy by manipulating nucleotide pools.
Main Methods:
- Liquid chromatography-mass spectrometry to measure cellular ribonucleotides (RNs) and deoxyribonucleotides (dRNs).
- Click chemistry to monitor RNA and DNA synthesis.
- Cell viability assays to determine IC50 values.
Main Results:
- RDV treatment led to S-phase arrest and suppressed RNA/DNA synthesis in BEAS-2B cells.
- Significant increases in nucleotide abundance were observed, with notable imbalance between purine and pyrimidine ribonucleotides.
- Potential inhibition of CTP synthase by RDV was indicated by disequilibrium between CTP and cytidine monophosphate.
Conclusions:
- RDV perturbs cellular nucleotide pools, inhibiting viral RNA replication.
- Targeting and exacerbating nucleotide pool imbalances could enhance RDV efficacy for COVID-19 treatment.
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