CD73 on T Cells Orchestrates Cardiac Wound Healing After Myocardial Infarction by Purinergic Metabolic Reprogramming
CD73 on T Cells Orchestrates Cardiac Wound Healing After Myocardial Infarction by Purinergic Metabolic Reprogramming
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DOI：
10.1161/circulationaha.116.023365
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发表时间：
2017-07-18
期刊：
影响因子：
37.8
通讯作者：
Schrader, Juergen
中科院分区：
文献类型：
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作者：
Borg, Nadine;Alter, Christina;Schrader, Juergen
背景：心肌梗死后T细胞是正常愈合所必需的。然而，它们的有益作用机制尚不清楚。损伤细胞释放的致炎危险信号ATP被胞外核苷酸CD39和CD73降解为抗炎介质腺苷。在这里，我们研究了T细胞产生的CD73衍生的腺苷在缺血/再灌流后心脏重构中的作用，并确定了其作用机制。方法：在全脑CD73(-/-)和CD4-CD73(-/-)小鼠中诱导心肌缺血(50min后再灌注)。对组织损伤、T细胞嘌呤能信号、细胞因子和心功能(4周9.4T磁共振断层扫描)进行分析。结果：CD73(-/-)小鼠的功能参数变化与CD73整体基因敲除(KO)小鼠相同。T细胞在基因(定量聚合酶链式反应)和蛋白质(酶活性)水平显著上调，关键转运蛋白和酶(连接蛋白43、连接蛋白37、pannin-1、平衡核苷转运蛋白1、CD39、CD73、外核苷酸焦磷酸酶/磷酸二酯酶1和3、CD157、CD38)可加速释放和水解ATP、cAMP、AMP和NAD为腺苷。令人惊讶的是，T细胞(来自CD39(-/-)小鼠)上CD39的缺乏并没有改变ATP的水解，并且很可能涉及焦磷酸酶(外核苷酸焦磷酸酶/磷酸二酯酶1和3)。循环T细胞主要表达A(2a)受体(A(2a)R)转录本。心肌梗死后，心脏T细胞和髓样细胞均有A(2b)受体(A(2b)R)转录的诱导。因此，A(2a)R和A(2b)R信号可能参与了心肌梗死后的心肌反应。就T细胞而言，这与CD73缺乏时促炎症和促纤维化细胞因子(白介素2、干扰素和白介素17)的加速分泌有关。外周淋巴结T细胞产生的细胞因子受到A(2a)R激活(CGS-21680)的抑制。A(2b)R激动剂Bay 60-6583表现出非靶点效应。腺苷受体激动剂NECA抑制干扰素-γ和刺激白介素6的产生，两者均可被A(2b)R拮抗剂(PSB-603)拮抗。结论：T细胞表面CD73在心肌梗死后心脏创面愈合过程中起重要作用。其潜在机制涉及ATP/NAD和AMP水解酶的显著增加，其主要原因是焦磷酸酶和CD73的上调。我们还定义了A(2b)R/A(2a)R介导的由T细胞来源的CD73介导的促炎症/促纤维化细胞因子的金盏花环反馈抑制。
BACKGROUND: T cells are required for proper healing after myocardial infarction. The mechanism of their beneficial action, however, is unknown. The proinflammatory danger signal ATP, released from damaged cells, is degraded by the ectonucleotidases CD39 and CD73 to the anti-inflammatory mediator adenosine. Here, we investigate the contribution of CD73-derived adenosine produced by T cells to cardiac remodeling after ischemia/reperfusion and define its mechanism of action.METHODS: Myocardial ischemia (50 minutes followed by reperfusion) was induced in global CD73(-/-) and CD4-CD73(-/-) mice. Tissue injury, T-cell purinergic signaling, cytokines, and cardiac function (magnetic resonance tomography at 9.4 T over 4 weeks) were analyzed.RESULTS: Changes in functional parameters of CD4-CD73(-/-) mice were identical to those in global CD73 knockouts (KOs). T cells infiltrating the injured heart significantly upregulated at the gene (quantitative polymerase chain reaction) and protein (enzymatic activity) levels critical transporters and enzymes (connexin43, connexin37, pannexin-1, equilibrative nucleoside transporter 1, CD39, CD73, ecto-nucleotide pyrophosphatase/phosphodiesterases 1 and 3, CD157, CD38) for the accelerated release and hydrolysis of ATP, cAMP, AMP, and NAD to adenosine. It is surprising that a lack of CD39 on T cells (from CD39(-/-) mice) did not alter ATP hydrolysis and very likely involves pyrophosphatases (ecto-nucleotide pyrophosphatase/phosphodiesterases 1 and 3). Circulating T cells predominantly expressed A(2a) receptor (A(2a)R) transcripts. After myocardial infarction, A(2b) receptor (A(2b)R) transcription was induced in both T cells and myeloid cells in the heart. Thus, A(2a)R and A(2b)R signaling may contribute to myocardial responses after myocardial infarction. In the case of T cells, this was associated with an accelerated secretion of proinflammatory and profibrotic cytokines (interleukin-2, interferon-gamma, and interleukin-17) when CD73 was lacking. Cytokine production by T cells from peripheral lymph nodes was inhibited by A(2a)R activation (CGS-21680). The A(2b)R agonist BAY 60-6583 showed off-target effects. The adenosine receptor agonist NECA inhibited interferon-gamma and stimulated interleukin-6 production, each of which was antagonized by a specific A(2b)R antagonist (PSB-603).CONCLUSIONS: This work demonstrates that CD73 on T cells plays a crucial role in the cardiac wound healing process after myocardial infarction. The underlying mechanism involves a profound increase in the hydrolysis of ATP/NAD and AMP, resulting primarily from the upregulation of pyrophosphatases and CD73. We also define A(2b)R/A(2a)R-mediated autacoid feedback inhibition of proinflammatory/profibrotic cytokines by T cell-derived CD73.