Department of Pathology, Michigan Medicine, University of Michigan, Ann Arbor, Michigan, United States of America.
Mary H. Weiser Food Allergy Center, Michigan Medicine, University of Michigan, Ann Arbor, Michigan, United States of America.
PLoS Pathog. 2020 Feb 27;16(2):e1008319. doi: 10.1371/journal.ppat.1008319. eCollection 2020 Feb.
Respiratory syncytial virus (RSV) is the major cause of lower respiratory tract infection in children worldwide. Sirtuin 1 (SIRT1), a NAD+ dependent deacetylase, has been associated with induction of autophagy, reprogramming cellular metabolism, and regulating immune mediators. In this study, we investigated the role of SIRT1 in bone marrow dendritic cell (BMDC) function during RSV infection. SIRT1 deficient (SIRT1 -/-) BMDC showed a defect in mitochondrial membrane potential (Δ⍦m) that worsens during RSV infection. This defect in Δ⍦m caused the generation of elevated levels of reactive oxygen species (ROS). Furthermore, the oxygen consumption rate (OCR) was reduced as assessed in Seahorse assays, coupled with lower levels of ATP in SIRT1-/- DC. These altered responses corresponded to altered innate cytokine responses in the SIRT1-/- DC in response to RSV infection. Reverse Phase Protein Array (RPPA) functional proteomics analyses of SIRT1-/- and WT BMDC during RSV infection identified a range of differentially regulated proteins involved in pathways that play a critical role in mitochondrial metabolism, autophagy, oxidative and ER stress, and DNA damage. We identified an essential enzyme, acetyl CoA carboxylase (ACC1), which plays a central role in fatty acid synthesis and had significantly increased expression in SIRT1-/- DC. Blockade of ACC1 resulted in metabolic reprogramming of BMDC that ameliorated mitochondrial dysfunction and reduced pathologic innate immune cytokines in DC. The altered DC responses attenuated Th2 and Th17 immunity allowing the appropriate generation of anti-viral Th1 responses both in vitro and in vivo during RSV infection thus reducing the enhanced pathogenic responses. Together, these studies identify pathways critical for appropriate DC function and innate immunity that depend on SIRT1-mediated regulation of metabolic processes.
呼吸道合胞病毒(RSV)是全球儿童下呼吸道感染的主要原因。Sirtuin 1(SIRT1)是一种 NAD+依赖性去乙酰化酶,与自噬的诱导、细胞代谢的重编程和免疫调节剂的调节有关。在这项研究中,我们研究了 SIRT1 在 RSV 感染期间骨髓树突状细胞(BMDC)功能中的作用。SIRT1 缺陷(SIRT1-/-)BMDC 显示线粒体膜电位(Δ⍦m)缺陷,在 RSV 感染过程中恶化。这种 Δ⍦m 缺陷导致活性氧(ROS)水平升高。此外,在 Seahorse 测定中评估的耗氧量(OCR)降低,同时 SIRT1-/- DC 中的 ATP 水平降低。这些改变的反应与 SIRT1-/- DC 对 RSV 感染的固有细胞因子反应的改变相对应。在 RSV 感染期间,对 SIRT1-/-和 WT BMDC 的反向蛋白质阵列(RPPA)功能蛋白质组学分析确定了一系列差异调节的蛋白质,这些蛋白质涉及在线粒体代谢、自噬、氧化和 ER 应激以及 DNA 损伤中起关键作用的途径。我们鉴定了一种必需酶,乙酰辅酶 A 羧化酶(ACC1),它在脂肪酸合成中起核心作用,并且在 SIRT1-/- DC 中表达显著增加。ACC1 阻断导致 BMDC 的代谢重编程,改善线粒体功能障碍并减少 DC 中的病理性固有免疫细胞因子。改变的 DC 反应减弱了 Th2 和 Th17 免疫,从而在 RSV 感染期间在体外和体内都适当产生抗病毒 Th1 反应,从而减轻增强的致病反应。总之,这些研究确定了适当的 DC 功能和固有免疫所必需的途径,这些途径依赖于 SIRT1 介导的代谢过程调节。