The role of N-Acylethanolamine Acid Amidase in the replication and maturation of ss(+) RNA viruses.

Single-stranded positive-sense RNA viruses are known to manipulate host lipids to create membrane vesicles within their replicate. In this study, we investigated the role of N-Acylethanolamine acid amidase (NAAA) in viral replication and its potential as a target for antiviral therapy. Using CRISPR/Cas9 gene editing and pharmacological NAAA inhibitors, we assessed the impact of NAAA on the replication of Flaviviruses, including Zika virus (ZIKV), Yellow Fever virus, and Dengue virus, as well as the new Coronavirus SARS-CoV-2. Our findings demonstrated that NAAA ablation and pharmacological inhibition with ARN726 led to a significant reduction in ZIKV replication in A549 cells. The potent NAAA inhibitor, ARN726, also effectively inhibited ZIKV replication in human neuronal stem cells and impaired ZIKV maturation by disrupting the cleavage of prM by Furin protease. Additionally, NAAA inhibitors showed promising antiviral activity against Yellow Fever and Dengue viruses, resulting in a decrease in viral replication. Furthermore, our study revealed that NAAA inhibition significantly reduced SARS-CoV-2 replication in both 2D cell culture and a 3D model using Human Precision-Cut Lung slides. Moreover, we also observed that NAAA inhibition displayed anti-inflammatory properties in human peripheral blood mononuclear cells (PBMC) derived from healthy donors. The treatment with NAAA inhibitor ARN726 resulted in a significant reduction in the release of pro-inflammatory cytokines TNF-α and IL-8, indicating its potential to mitigate the inflammation associated with viral infections. In conclusion, our study highlights the potential of NAAA inhibitors as a dual approach to combat viral infections caused by Flaviviruses and SARS-CoV-2, while also reducing inflammation associated with these infections.