Fu Jiayao, Shi Huan, Wang Baoli, Zhan Tianle, Shao Yanxiong, Ye Lei, Wu Shufeng, Yu Chuangqi, Zheng Lingyan
Department of Oral Surgery, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Clinical Research Center of Oral Disease, Shanghai, China; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai, China.
Department of Oral Surgery, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Clinical Research Center of Oral Disease, Shanghai, China; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai, China.
J Autoimmun. 2020 Feb;107:102358. doi: 10.1016/j.jaut.2019.102358. Epub 2019 Nov 19.
The hyperproliferation and hyperactivation of CD4 T cells in salivary gland tissue is a hallmark of Sjögren's syndrome (SS). However, the role of long noncoding RNAs (lncRNAs) in the pathological process of SS and CD4 T cell activation has not been fully elucidated. Here, we reported that lncRNA PVT1 was involved in the glycolytic metabolism reprogramming and proliferation upon CD4 T cell activation. Expression of PVT1 was positively related with CD4 T cell activation both in SS patients and Ex vivo antigen simulation. Depletion of PVT1 decreased the proliferation of murine CD4 T cells and Jurkat T cells upon activation. We also showed that expression of the transcription factor Myc is regulated by PVT1 under antigen simulation. Depletion of PVT1 significantly decreased the expression of glycolytic genes, as well as several pivotal glycolytic proteins that were directly transcribed by Myc. Measurement of glucose content and lactate secretion indicated a defected lactate secretion and glucose uptake in PVT1-depleted T cells. Additionally, the real-time extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) measurement also affirmed that PVT1 maintains glycolytic levels, glycolytic capacity under stress and ECAR/OCR ratios during T cell activation. Polarizing assays indicate that PVT1 depletion defected the function of Th1 effector cells as well as down-regulated Myc expression and glycolytic levels. Furthermore, we observed increased glycolytic levels in CD4 T cells from SS-like NOD/Ltj mice. Treatment with 2-deoxy-d-glucose (2-DG), an inhibitor of glycolysis, significantly decreased the extent of lymphocyte infiltration and CD4 T cell numbers and attenuated the defect of salivary flow in the lesioned submandibular glands of NOD/Ltj mice. Thus, our study demonstrated that lncRNA PVT1, which was upregulated in the CD4T cells of SS patients, could maintain the expression of Myc, thus controlling the proliferation and effector functions of CD4 T cells through regulating the reprogramming of glycolysis. Inhibition of glycolysis could attenuate the proliferation of CD4 T cells and the SS-like autoimmune response. Our study provides a novel mechanistic function of lncRNA PVT1 in the pathogenesis of SS.
唾液腺组织中CD4 T细胞的过度增殖和过度活化是干燥综合征（SS）的一个标志。然而，长链非编码RNA（lncRNA）在SS病理过程和CD4 T细胞活化中的作用尚未完全阐明。在此，我们报道lncRNA PVT1参与CD4 T细胞活化后的糖酵解代谢重编程和增殖。在SS患者以及体外抗原模拟中，PVT1的表达均与CD4 T细胞活化呈正相关。PVT1的缺失降低了活化后小鼠CD4 T细胞和Jurkat T细胞的增殖。我们还表明，在抗原模拟下，转录因子Myc的表达受PVT1调控。PVT1的缺失显著降低了糖酵解基因以及由Myc直接转录的几种关键糖酵解蛋白的表达。葡萄糖含量和乳酸分泌的测定表明，PVT1缺失的T细胞中乳酸分泌和葡萄糖摄取存在缺陷。此外，实时细胞外酸化率（ECAR）和耗氧率（OCR）的测量也证实，PVT1在T细胞活化过程中维持糖酵解水平、应激状态下的糖酵解能力以及ECAR/OCR比值。极化分析表明，PVT1的缺失损害了Th1效应细胞的功能，并下调了Myc表达和糖酵解水平。此外，我们观察到类似SS的NOD/Ltj小鼠的CD4 T细胞中糖酵解水平升高。用糖酵解抑制剂2-脱氧-D-葡萄糖（2-DG）处理显著降低了淋巴细胞浸润程度和CD4 T细胞数量，并减轻了NOD/Ltj小鼠病变下颌下腺唾液分泌的缺陷。因此，我们的研究表明，在SS患者的CD4 T细胞中上调的lncRNA PVT1可以维持Myc的表达，从而通过调节糖酵解重编程来控制CD4 T细胞的增殖和效应功能。抑制糖酵解可以减弱CD4 T细胞的增殖和类似SS的自身免疫反应。我们的研究为lncRNA PVT1在SS发病机制中的新机制功能提供了依据。
