Lipid droplets fuel SARS-CoV-2 replication and production of inflammatory mediators.
Lipid droplets fuel SARS-CoV-2 replication and production of inflammatory mediators.
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DOI：
10.1371/journal.ppat.1009127
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发表时间：
2020-12
期刊：
影响因子：
6.7
通讯作者：
Bozza PT
中科院分区：
文献类型：
--
作者：
Dias SSG;Soares VC;Ferreira AC;Sacramento CQ;Fintelman-Rodrigues N;Temerozo JR;Teixeira L;Nunes da Silva MA;Barreto E;Mattos M;de Freitas CS;Azevedo-Quintanilha IG;Manso PPA;Miranda MD;Siqueira MM;Hottz ED;Pão CRR;Bou-Habib DC;Barreto-Vieira DF;Bozza FA;Souza TML;Bozza PT
病毒是专性细胞内寄生虫，利用宿主代谢机制来满足其生物合成需要。因此，确定病毒复制所必需的宿主途径可能导致治疗干预的潜在目标。SARS-CoV-2支持其在宿主细胞内复制的机制和途径尚不完全清楚。脂滴是一种具有脂质代谢、能量稳态和细胞内运输等功能的细胞器，在感染和炎症中具有多种作用。在这里，我们描述了来自COVID-19患者的单核细胞与SARS-CoV-2阴性供者相比，LD积累增加。在体外，通过检测单核细胞中CD36、SREBP-1、PPARγ和DGAT-1的表达，观察到SARS-CoV-2感染可调节脂质合成和摄取途径，并触发不同人类细胞系中LD的形成。在感染的Vero细胞中发现lld与SARS-CoV-2蛋白和双链rna密切相关。对SARS-CoV-2感染的Vero细胞的电镜分析显示，病毒颗粒与ld共定位，提示ld可能作为组装平台。A922500抑制DGAT-1对LD形成的药理学调节显著抑制了SARS-CoV-2的复制，并减少了促炎反应介质的产生。总之，我们证明了脂质代谢重编程和LD形成在SARS-CoV-2复制和发病机制中的重要作用，为COVID-19的治疗策略开辟了新的机会。尽管在了解SARS-CoV2诱导疾病的机制和制定控制COVID-19大流行的策略方面做出了巨大的科学努力，但SARS-CoV2生物学和发病机制的许多方面仍然难以理解。一些RNA病毒能够调节宿主脂质代谢并招募ld以增强其复制/颗粒组装能力，其机制因病毒和感染的宿主细胞而异。SARS-CoV-2探索的支持其在宿主细胞内复制的机制和途径在很大程度上仍然未知。在这里，我们证明脂滴（ld）参与SARS-CoV2感染，有利于病毒复制和增强炎症介质的产生。SARS-CoV2感染增加了脂质代谢调节关键蛋白的表达和每个细胞的ld数量。此外，我们在感染细胞中发现了SARS-CoV2和/或其与lld相关的成分，提示lld是作为复制室的一部分被招募的。此外，药物抑制DGAT-1 （LD形成的关键酶）可减少SARS-CoV2的复制、炎症介质的产生和细胞死亡。我们的发现有助于揭示SARS-CoV-2利用细胞代谢和细胞器协调病毒复制周期和宿主免疫不同步骤的复杂机制，为SARS-CoV-2抗病毒药物的开发开辟了新的视角。
Viruses are obligate intracellular parasites that make use of the host metabolic machineries to meet their biosynthetic needs. Thus, identifying the host pathways essential for the virus replication may lead to potential targets for therapeutic intervention. The mechanisms and pathways explored by SARS-CoV-2 to support its replication within host cells are not fully known. Lipid droplets (LD) are organelles with major functions in lipid metabolism, energy homeostasis and intracellular transport, and have multiple roles in infections and inflammation. Here we described that monocytes from COVID-19 patients have an increased LD accumulation compared to SARS-CoV-2 negative donors. In vitro, SARS-CoV-2 infection were seen to modulate pathways of lipid synthesis and uptake as monitored by testing for CD36, SREBP-1, PPARγ, and DGAT-1 expression in monocytes and triggered LD formation in different human cell lines. LDs were found in close apposition with SARS-CoV-2 proteins and double-stranded (ds)-RNA in infected Vero cells. Electron microscopy (EM) analysis of SARS-CoV-2 infected Vero cells show viral particles colocalizing with LDs, suggestive that LDs might serve as an assembly platform. Pharmacological modulation of LD formation by inhibition of DGAT-1 with A922500 significantly inhibited SARS-CoV-2 replication as well as reduced production of mediators pro-inflammatory response. Taken together, we demonstrate the essential role of lipid metabolic reprograming and LD formation in SARS-CoV-2 replication and pathogenesis, opening new opportunities for therapeutic strategies to COVID-19. In spite of the enormous scientific efforts to understand mechanisms of SARS-CoV2-induced disease and to develop strategies to control COVID-19 pandemic, many aspects of SARS-CoV2 biology and pathogenesis remain elusive. Several RNA viruses are able to modulate the host lipid metabolism and to recruit LDs to enhance their replication/particle assembling capacity through mechanisms that vary according to the virus and the host cell infected. The mechanisms and pathways explored by SARS-CoV-2 to support its replication within host cells are still largely unknown. Here we demonstrated that lipid droplets (LDs) participate in SARS-CoV2 infection favoring virus replication and heightening inflammatory mediator production. SARS-CoV2 infection increased the expression of key proteins in the regulation of lipid metabolism and the amounts of LDs per cell. In addition, we have found SARS-CoV2 and/or its components associated with LDs in infected cells, suggestive that LDs are recruited as part of replication compartment. Moreover, pharmacological inhibition of DGAT-1, a key enzyme for LD formation, reduces SARS-CoV2 replication, inflammatory mediator production and cell death. Our findings contribute to unveil the complex mechanism by which SARS-CoV-2 make use of cellular metabolism and organelles to coordinate different steps of the viral replication cycle and host immunity, opening new perspectives for SARS-CoV2 antiviral development.
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DOI：
10.1590/0074-02760150433
发表时间：
2016-06-01
期刊：
Memórias do Instituto Oswaldo Cruz
影响因子：
--
作者：
Barth, Ortrud Monika;Silva, Marcos Alexandre Nunes da;Barreto-Vieira, Debora Ferreira
通讯作者：
Barreto-Vieira, Debora Ferreira
影响因子：
5.4
作者：
Carvalho, Filomena A.;Carneiro, Fabiana A.;Santos, Nuno C.
通讯作者：
Santos, Nuno C.
影响因子：
5.9
作者：
Crawford, Sue E.;Desselberger, Ulrich
通讯作者：
Desselberger, Ulrich
DOI：
10.1016/j.bbalip.2013.10.008
发表时间：
2014-01-01
影响因子：
4.8
作者：
Almeida, Patricia E.;Roque, Natalia R.;Bozza, Patricia T.
通讯作者：
Bozza, Patricia T.
影响因子：
15.9
作者：
Febbraio, M;Hajjar, DP;Silverstein, RL
通讯作者：
Silverstein, RL