Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate.
Glutaminase 1 regulates the release of extracellular vesicles during neuroinflammation through key metabolic intermediate alpha-ketoglutarate.
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DOI：
10.1186/s12974-018-1120-x
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发表时间：
2018-03-14
影响因子：
9.3
通讯作者：
Zheng JC
中科院分区：
文献类型：
--
作者：
Wu B;Liu J;Zhao R;Li Y;Peer J;Braun AL;Zhao L;Wang Y;Tong Z;Huang Y;Zheng JC
细胞外囊泡（EV）在中枢神经系统的细胞间通讯中是重要的，并且它们的释放在神经炎症期间增加。我们以前的数据表明，在HIV-1感染和神经胶质细胞免疫激活期间，EV的释放增加。然而，感染和炎症增加EV释放的分子机制仍然未知。在目前的研究中，我们研究了辅酶A酶1（GLS 1）介导的辅酶A胺解和关键代谢中间体α-酮戊二酸的产生对EV释放的作用。人单核细胞衍生的巨噬细胞原代培养物和BV 2小胶质细胞系用于代表CNS中的先天免疫细胞。透射电子显微镜，纳米粒子跟踪分析，和Western印迹被用来确定EV的调节。使用腺病毒载体在体外和转基因小鼠模型在体内进行GLS 1过表达。通过ANOVA对数据进行统计学评价，然后对配对观察结果进行Bonferroni事后检验。我们的数据显示，在GLS 1过表达的HeLa细胞中，EV的释放增加。在HIV-1感染的巨噬细胞和免疫激活的小胶质细胞BV 2细胞中，用双-2-（5-苯乙酰氨基-1，2，4-噻二唑-2-基）乙基硫醚（BPTES）或CB 839（两种特异性GLS抑制剂）处理显著降低EV释放，表明GLS 1在EV释放中的关键作用。此外，添加α-酮戊二酸或神经酰胺可在BPTES治疗期间挽救EV释放，表明α-酮戊二酸和神经酰胺是GLS抑制剂的关键下游效应物。这些发现与GLS 1转基因小鼠脑组织的研究进一步证实。GLS 1转基因小鼠的EV水平显著高于对照小鼠，表明GLS 1增加了体内EV释放。这些结果表明，GLS 1介导的β-氨基分解及其下游α-酮戊二酸的产生在调节HIV-1感染和免疫激活过程中EV的释放中是必不可少的。这些新的机制调节可能有助于理解谷氨酰胺代谢如何在神经炎症期间塑造EV生物发生和释放。本文的在线版本（10.1186/s12974-018-1120-x）包含补充材料，可供授权用户使用。
Extracellular vesicles (EVs) are important in the intercellular communication of the central nervous system, and their release is increased during neuroinflammation. Our previous data demonstrated an increased release of EVs during HIV-1 infection and immune activation in glial cells. However, the molecular mechanism by which infection and inflammation increase EV release remains unknown. In the current study, we investigated the role of glutaminase 1 (GLS1)-mediated glutaminolysis and the production of a key metabolic intermediate α-ketoglutarate on EV release. Human monocyte-derived macrophage primary cultures and a BV2 microglia cell line were used to represent the innate immune cells in the CNS. Transmission electron microscopy, nanoparticle tracking analysis, and Western blots were used to determine the EV regulation. GLS1 overexpression was performed using an adenovirus vector in vitro and transgenic mouse models in vivo. Data were evaluated statistically by ANOVA, followed by the Bonferroni post-test for paired observations. Our data revealed an increased release of EVs in GLS1-overexpressing HeLa cells. In HIV-1-infected macrophages and immune-activated microglia BV2 cells, treatment with bis-2-(5-phenylacetamido-1,2,4-thiadiazol-2-yl)ethyl sulfide (BPTES) or CB839, two specific GLS inhibitors, significantly decreased EV release, suggesting a critical role of GLS1 in EV release. Furthermore, addition of α-ketoglutarate or ceramide rescued EV release during BPTES treatment, implicating α-ketoglutarate and ceramide as critical downstream effectors for GLS inhibitors. These findings were further corroborated with the investigation of brain tissues in GLS1-transgenic mice. The EV levels were significantly higher in GLS1 transgenic mice than those in control mice, suggesting that GLS1 increases EV release in vivo. These findings suggest that GLS1-mediated glutaminolysis and its downstream production of α-ketoglutarate are essential in regulating EV release during HIV-1 infection and immune activation. These new mechanistic regulations may help understand how glutamine metabolism shapes EV biogenesis and release during neuroinflammation. The online version of this article (10.1186/s12974-018-1120-x) contains supplementary material, which is available to authorized users.
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影响因子：
4.2
作者：
Dinkins MB;Dasgupta S;Wang G;Zhu G;Bieberich E
通讯作者：
Bieberich E
影响因子：
--
作者：
Erickson JW;Cerione RA
通讯作者：
Cerione RA
影响因子：
4.7
作者：
Delint-Ramirez, Ilse;Salcedo-Tello, Pamela;Bermudez-Rattoni, Federico
通讯作者：
Bermudez-Rattoni, Federico
影响因子：
5.7
作者：
Gross, Matt I.;Demo, Susan D.;Bennett, Mark K.
通讯作者：
Bennett, Mark K.
影响因子：
3.3
作者：
Jiang, ZG;Piggee, C;Markey, SP
通讯作者：
Markey, SP