Understanding the fundamental biochemical and metabolic requirements for the replication of coronaviruses within infected cells is of notable interest for the development of broad-based therapeutic strategies, given the likelihood of emergence of new pandemic-potential virus species, as well as future variants of SARS-CoV-2. Here we demonstrate members of the glutaminase family of enzymes (GLS and GLS2), which catalyze the hydrolysis of glutamine to glutamate (i.e., the first step in glutamine metabolism), play key roles in coronavirus replication in host cells. Using a range of human seasonal and zoonotic coronaviruses, we show three examples where GLS expression increases during coronavirus infection of host cells, and another where GLS2 is upregulated. The viruses hijack the metabolic machinery responsible for glutamine metabolism to generate the building blocks for biosynthetic processes and satisfy the bioenergetic requirements demanded by the 'glutamine addiction' of virus-infected cells. We demonstrate that genetic silencing of glutaminase enzymes reduces coronavirus infection and that newer members of two classes of allosteric inhibitors targeting these enzymes, designated as SU1, a pan-GLS/GLS2 inhibitor, and UP4, a specific GLS inhibitor, block viral replication in epithelial cells. Moreover, treatment of SARS-CoV-2 infected K18-human ACE2 transgenic mice with SU1 resulted in their complete survival compared to untreated control animals, which succumbed within 10 days post-infection. Overall, these findings highlight the importance of glutamine metabolism for coronavirus replication in human cells and mice and show that glutaminase inhibitors can block coronavirus infection and thereby may represent a novel class of broad-based anti-viral drug candidates.

中文翻译：

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谷氨酰胺代谢对于冠状病毒在宿主细胞和小鼠中的复制至关重要。


鉴于可能出现新的大流行潜在病毒物种以及 SARS-CoV-2 的未来变体，了解冠状病毒在感染细胞内复制的基本生化和代谢要求对于制定基础广泛的治疗策略具有重要意义。在这里，我们展示了谷氨酰胺酶家族（GLS 和 GLS2）的成员，它们催化谷氨酰胺水解为谷氨酸（即谷氨酰胺代谢的第一步），在宿主细胞中的冠状病毒复制中起关键作用。使用一系列人类季节性和人畜共患冠状病毒，我们展示了三个例子，其中 GLS 表达在冠状病毒感染宿主细胞期间增加，另一个例子 GLS2 上调。病毒劫持了负责谷氨酰胺代谢的代谢机制，以产生生物合成过程的组成部分，并满足病毒感染细胞的“谷氨酰胺成瘾”所要求的生物能量需求。我们证明谷氨酰胺酶的遗传沉默减少了冠状病毒感染，并且针对这些酶的两类变构抑制剂的新成员，称为 SU1（一种泛 GLS/GLS2 抑制剂）和 UP4（一种特异性 GLS 抑制剂），阻断了病毒在上皮细胞中的复制。此外，与未治疗的对照动物相比，用 SU1 治疗 SARS-CoV-2 感染的 K18-人 ACE2 转基因小鼠导致它们在感染后 10 天内死亡，从而完全存活。总体而言，这些发现强调了谷氨酰胺代谢对冠状病毒在人类细胞和小鼠中复制的重要性，并表明谷氨酰胺酶抑制剂可以阻断冠状病毒感染，因此可能代表了一类新型基础广泛的抗病毒候选药物。