Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.
Targeting Glutamine Metabolism Ameliorates Autoimmune Hepatitis via Inhibiting T Cell Activation and Differentiation.
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靶向谷氨酰胺代谢通过抑制 T 细胞激活和分化来改善自身免疫性肝炎。
DOI：
10.3389/fimmu.2022.880262
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发表时间：
2022
影响因子：
7.3
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中科院分区：
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自身免疫性肝炎（AIH）是由针对肝细胞的T细胞介导的级联反应介导的，肝脏内持续的炎症最终可导致肝硬化。靶向谷氨酰胺代谢对T细胞活化和分化具有影响。然而，谷氨酰胺代谢阻断对AIH的影响尚不清楚。本研究采用谷氨酰胺酶拮抗剂6-重氮-5-氧代-L-正亮氨酸（DON）和其前体药物2-（2-氨基-4-甲基戊酰胺基）-DON（JHU083）分别进行体外和体内试验，探讨其对AIH小鼠模型谷氨酰胺代谢阻断的潜在治疗作用及其分子机制。在给予伴刀豆球蛋白A（ConA）之前，用JHU083或溶剂治疗AIH小鼠，并检查疾病严重程度。然后检测来自溶媒-WT、JHU083-AIH和溶媒-AIH小鼠的T细胞的活化和分化[包括Th1/Th17细胞和细胞毒性T淋巴细胞（CTL）]。此外，在体外T细胞的活化和分化进行了测量，使用分离的脾细胞刺激刀豆蛋白A与或没有DON。采用流式细胞术、qRT-PCR和ELISA检测T细胞的活化和分化。Western blotting检测mTOR和70 kDa核糖体蛋白S6激酶（P70 S6K）的磷酸化水平。JHU083和DON在体内外均能显著抑制T细胞活化，抑制Th1/Th17细胞和CTL的分化。此外，我们证明谷氨酰胺代谢阻断通过降低氨基酸转运蛋白溶质载体家族7成员5（SLC7A5）的mRNA表达和减轻mTOR信号转导的激活来抑制T细胞活化和分化。我们证明，针对谷氨酰胺代谢是AIH和其他T细胞介导疾病患者的潜在新治疗策略。从机制上讲，我们证明谷氨酰胺代谢阻断抑制T细胞活化，抑制Th1/Th17细胞和CTL的分化。
Autoimmune hepatitis (AIH) is mediated by a cascade of T cell-mediated events directed at liver cells and persistent inflammation within the liver can eventually result in liver cirrhosis. Targeting glutamine metabolism has an impact on T cell activation and differentiation. However, the effect of glutamine metabolism blocking upon AIH remains unknown. We use glutaminase antagonist 6-diazo-5-oxo-L-norleucine (DON) for in vitro assays and its prodrug 2-(2-amino-4-methylpentanamido)-DON (JHU083) for in vivo assays to investigate the potential therapeutic effect and molecular mechanism of glutamine metabolism blocking in an AIH murine model. AIH mice were treated with JHU083 or vehicle before concanavalin A (ConA) administration, and disease severity was examined. Then activation and differentiation [including Th1/Th17 cells and cytotoxic T lymphocytes (CTL)] of T cells from Vehicle-WT, JHU083-AIH and Vehicle-AIH mice were tested. Furthermore, in vitro T cell activation and differentiation were measured using separated splenocytes stimulated with ConA with or without DON. The activation and differentiation of T cells were tested using flow cytometry, qRT-PCR and ELISA. Phosphorylation level of mammalian target of rapamycin (mTOR) and 70 kDa ribosomal protein S6 kinase (P70S6K) were examined by western blotting. JHU083 and DON significantly suppressed the activation of T cells and inhibited the differentiation of Th1/Th17 cells and CTL in vivo and in vitro. Besides, we demonstrated that glutamine metabolism blocking inhibited T cells activation and differentiation through decreasing the mRNA expression of amino acid transporter solute carrier family 7 member 5 (SLC7A5) and mitigating the activation of mTOR signaling. We proved that targeting glutamine metabolism represents a potential new treatment strategy for patients with AIH and other T cell-mediated disease. Mechanistically, we demonstrated that glutamine metabolism blocking inhibits T cells activation and suppresses the differentiation of Th1/Th17 cells and CTL.