PDF(Pubmed)
Abstract：
The global impact of emerging viral infections emphasizes the urgent need for effective broad-spectrum antivirals. The cellular organelle, lipid droplet (LD), is utilized by many types of viruses for replication, but its reduction does not affect cell survival. Therefore, LD is a potential target for developing broad-spectrum antivirals. In this study, we found that 2-bromopalmitate (2 BP), a previously defined palmitoylation inhibitor, depletes LD across all studied cell lines and exerts remarkable antiviral effects on different coronaviruses. We comprehensively utilized 2 BP, alongside other palmitoylation inhibitors such as cerulenin and 2-fluoro palmitic acid (2-FPA), as well as the enhancer palmostatin B and evaluated their impact on LD and the replication of human coronaviruses (hCoV-229E, hCoV-Oc43) and murine hepatitis virus (MHV-A59) at non-cytotoxic concentrations. While cerulenin and 2-FPA exhibited moderate inhibition of viral replication, 2 BP exhibited a much stronger suppressive effect on MHV-A59 replication, although they share similar inhibitory effects on palmitoylation. As expected, palmostatin B significantly enhanced viral replication, it failed to rescue the inhibitory effects of 2 BP, whereas it effectively counteracted the effects of cerulenin and 2-FPA. This suggests that the mechanism that 2 BP used to inhibit viral replication is beyond palmitoylation inhibition. Further investigations unveil that 2 BP uniquely depletes LDs, a phenomenon not exhibited by 2-FPA and cerulenin. Importantly, the depletion of LDs was closely associated with the inhibition of viral replication because the addition of oleic acid to 2 BP significantly rescued LD depletion and its inhibitory effects on MHV-A59. Our findings indicate that the inhibitory effects of 2 BP on viral replication primarily stem from LD disruption rather than palmitoylation inhibition. Intriguingly, fatty acid (FA) assays demonstrated that 2 BP reduces the FA level in mitochondria while concurrently increasing FA levels in the cytoplasm. These results highlight the crucial role of LDs in viral replication and uncover a novel biological activity of 2 BP. These insights contribute to the development of broad-spectrum antiviral strategies.
OBJECTIVE: In our study, we conducted a comparative investigation into the antiviral effects of palmitoylation inhibitors including 2-bromopalmitate (2-BP), 2-fluoro palmitic acid (2-FPA), and cerulenin. Surprisingly, we discovered that 2-BP has superior inhibitory effects on viral replication compared to 2-FPA and cerulenin. However, their inhibitory effects on palmitoylation were the same. Intrigued by this finding, we delved deeper into the underlying mechanism of 2-BP\'s potent antiviral activity, and we unveiled a novel biological activity of 2-BP: depletion of lipid droplets (LDs). Importantly, we also highlighted the crucial role of LDs in viral replication. Our insights shed new light on the antiviral mechanism of LD depletion paving the way for the development of broad-spectrum antiviral strategies by targeting LDs.
OBJECTIVE: In our study, we conducted a comparative investigation into the antiviral effects of palmitoylation inhibitors including 2-bromopalmitate (2-BP), 2-fluoro palmitic acid (2-FPA), and cerulenin. Surprisingly, we discovered that 2-BP has superior inhibitory effects on viral replication compared to 2-FPA and cerulenin. However, their inhibitory effects on palmitoylation were the same. Intrigued by this finding, we delved deeper into the underlying mechanism of 2-BP\'s potent antiviral activity, and we unveiled a novel biological activity of 2-BP: depletion of lipid droplets (LDs). Importantly, we also highlighted the crucial role of LDs in viral replication. Our insights shed new light on the antiviral mechanism of LD depletion paving the way for the development of broad-spectrum antiviral strategies by targeting LDs.
摘要：
新出现的病毒感染的全球影响强调迫切需要有效的广谱抗病毒药物。细胞器，脂滴（LD），被许多类型的病毒用于复制，但其减少并不影响细胞存活。因此,LD是开发广谱抗病毒药物的潜在目标。在这项研究中,我们发现2-溴棕榈酸酯（2BP），先前定义的棕榈酰化抑制剂，在所有研究的细胞系中消耗LD，并对不同的冠状病毒发挥显着的抗病毒作用。我们综合利用了2个BP，与其他棕榈酰化抑制剂一起，如天红素和2-氟棕榈酸（2-FPA），以及增强子palmostatinB，并评估了它们对LD和人冠状病毒复制的影响（hCoV-229E，hCoV-Oc43）和非细胞毒性浓度的鼠肝炎病毒（MHV-A59）。而cerulenin和2-FPA显示病毒复制的中度抑制，2BP对MHV-A59复制表现出更强的抑制作用，尽管它们对棕榈酰化具有相似的抑制作用。不出所料,palmostatinB显着增强病毒复制，它未能挽救2BP的抑制作用，而它有效地抵消了cerulenin和2-FPA的作用。这表明2BP用于抑制病毒复制的机制超出了棕榈酰化抑制。进一步的调查揭示了2BP独特地消耗了LDs，2-FPA和蓝蛋白未表现出的现象。重要的是,LDs的耗竭与病毒复制的抑制密切相关,因为向2BP中添加油酸显著挽救了LD耗竭及其对MHV-A59的抑制作用.我们的发现表明，2BP对病毒复制的抑制作用主要源于LD破坏，而不是棕榈酰化抑制。有趣的是，脂肪酸（FA）测定表明，2BP降低线粒体中的FA水平，同时增加细胞质中的FA水平。这些结果突出了LDs在病毒复制中的关键作用，并揭示了2BP的新生物活性。这些见解有助于广谱抗病毒策略的发展。
目的：在我们的研究中，我们对棕榈酰化抑制剂的抗病毒作用进行了比较研究，包括2-溴棕榈酸酯（2-BP），2-氟棕榈酸(2-FPA),和cerulenin。令人惊讶的是,我们发现2-BP对病毒复制的抑制作用优于2-FPA和cerulenin.然而,它们对棕榈酰化的抑制作用相同。对这一发现感兴趣，我们深入研究了2-BP有效抗病毒活性的潜在机制，我们揭示了2-BP的新生物活性：脂滴（LDs）的消耗。重要的是,我们还强调了LD在病毒复制中的关键作用.我们的见解对LD耗竭的抗病毒机制有了新的认识，为通过靶向LD开发广谱抗病毒策略铺平了道路。
目的：在我们的研究中，我们对棕榈酰化抑制剂的抗病毒作用进行了比较研究，包括2-溴棕榈酸酯（2-BP），2-氟棕榈酸(2-FPA),和cerulenin。令人惊讶的是,我们发现2-BP对病毒复制的抑制作用优于2-FPA和cerulenin.然而,它们对棕榈酰化的抑制作用相同。对这一发现感兴趣，我们深入研究了2-BP有效抗病毒活性的潜在机制，我们揭示了2-BP的新生物活性：脂滴（LDs）的消耗。重要的是,我们还强调了LD在病毒复制中的关键作用.我们的见解对LD耗竭的抗病毒机制有了新的认识，为通过靶向LD开发广谱抗病毒策略铺平了道路。
