A Proteomics-Based Assessment of Inflammation Signatures in Endotoxemia.
A Proteomics-Based Assessment of Inflammation Signatures in Endotoxemia.
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基于蛋白质组学的内毒素血症炎症特征评估。
DOI：
10.1074/mcp.ra120.002305
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Mayr M
中科院分区：
文献类型：
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作者：
Burnap SA;Mayr U;Shankar-Hari M;Cuello F;Thomas MR;Shah AM;Sabroe I;Storey RF;Mayr M
我们之前已经证明，血浆五聚体 3 (PTX3) 的多聚体可以预测脓毒症患者的生存情况。为了表征脓毒症中血浆蛋白变化的释放动力学和细胞来源，从注射低剂量内毒素（脂多糖 [LPS]）的健康志愿者（n = 10；三个时间点）获得系列样品，并使用数据独立的采集 MS 进行分析。将人血浆蛋白质组反应与 LPS 诱导的小鼠内毒素血症模型进行比较。对人血浆的蛋白质组学分析揭示了中性粒细胞快速脱粒特征，随后急性期蛋白增加。注射 LPS 后，循环 PTX3 的变化与中性粒细胞衍生蛋白的增加相关。对小鼠血浆蛋白质组的时程分析显示，LPS 治疗后，多聚体 PTX3 呈时间依赖性增加，同时中性粒细胞衍生的髓过氧化物酶 (MPO) 也增加。在两种基因小鼠模型中探讨了氧化诱导的 PTX3 多聚化机制：MPO 全局敲除 (KO) 小鼠和 LysM Cre Nox2 KO 小鼠，其中 NADPH 氧化酶 2 (Nox2) 仅在骨髓细胞中缺乏。 Nox2 是负责中性粒细胞氧化爆发的酶。野生型和 MPO 或 LysM Cre Nox2 KO 小鼠之间血浆多聚体 PTX3 的增加没有显着差异。因此，PTX3可能已经以多聚体形式储存和释放。通过体内中性粒细胞耗竭和多重血管蛋白质组学，证实主动脉内的 PTX3 多聚体沉积具有中性粒细胞依赖性。对注射 LPS 的小鼠主动脉进行的蛋白质组学分析显示，PTX3 是上调最多的蛋白质，其中多聚体 PTX3 早在 LPS 后 2 小时就与其他中性粒细胞衍生的蛋白质一起沉积。总之，LPS 注射后多聚体 PTX3 的增加与血浆和主动脉中中性粒细胞相关蛋白的变化相关。 PTX3 的多聚化不需要 MPO 和髓系 Nox2；相反，中性粒细胞外渗是 LPS 诱导的多聚体 PTX3 在主动脉沉积的原因。 pentraxin-3 (PTX3) 多聚体的形成与脓毒症的结局有关。 PTX3 血浆水平与中性粒细胞衍生蛋白的增加相关。中性粒细胞衍生的髓过氧化物酶和 NADPH 氧化酶 2 不负责 PTX3 氧化。主动脉内 PTX3 多聚体沉积是中性粒细胞依赖性的。利用蛋白质组学方法绘制了人类和小鼠响应内毒素的循环蛋白质组变化。多聚体 pentraxin-3 (PTX3) 血浆和主动脉水平模拟了已知中性粒细胞来源的蛋白质响应内毒素而观察到的变化。遗传小鼠模型排除了髓过氧化物酶和 NADPH 氧化酶 2 作为 PTX3 氧化驱动因素的可能性；然而，中性粒细胞外渗会促进 PTX3 在脉管系统内沉积。结果揭示了全身炎症反应与多聚体 PTX3 的中性粒细胞依赖性血管沉积之间的联系。
We have previously shown that multimers of plasma pentraxin-3 (PTX3) were predictive of survival in patients with sepsis. To characterize the release kinetics and cellular source of plasma protein changes in sepsis, serial samples were obtained from healthy volunteers (n = 10; three time points) injected with low-dose endotoxin (lipopolysaccharide [LPS]) and analyzed using data-independent acquisition MS. The human plasma proteome response was compared with an LPS-induced endotoxemia model in mice. Proteomic analysis of human plasma revealed a rapid neutrophil degranulation signature, followed by a rise in acute phase proteins. Changes in circulating PTX3 correlated with increases in neutrophil-derived proteins following LPS injection. Time course analysis of the plasma proteome in mice showed a time-dependent increase in multimeric PTX3, alongside increases in neutrophil-derived myeloperoxidase (MPO) upon LPS treatment. The mechanisms of oxidation-induced multimerization of PTX3 were explored in two genetic mouse models: MPO global knock-out (KO) mice and LysM Cre Nox2 KO mice, in which NADPH oxidase 2 (Nox2) is only deficient in myeloid cells. Nox2 is the enzyme responsible for the oxidative burst in neutrophils. Increases in plasma multimeric PTX3 were not significantly different between wildtype and MPO or LysM Cre Nox2 KO mice. Thus, PTX3 may already be stored and released in a multimeric form. Through in vivo neutrophil depletion and multiplexed vascular proteomics, PTX3 multimer deposition within the aorta was confirmed to be neutrophil dependent. Proteomic analysis of aortas from LPS-injected mice returned PTX3 as the most upregulated protein, where multimeric PTX3 was deposited as early as 2 h post-LPS along with other neutrophil-derived proteins. In conclusion, the rise in multimeric PTX3 upon LPS injection correlates with neutrophil-related protein changes in plasma and aortas. MPO and myeloid Nox2 are not required for the multimerization of PTX3; instead, neutrophil extravasation is responsible for the LPS-induced deposition of multimeric PTX3 in the aorta. Multimer formation of pentraxin-3 (PTX3) has been linked to outcome in sepsis. PTX3 plasma levels have been correlated to increases in neutrophil-derived proteins. Neutrophil-derived myeloperoxidase and NADPH oxidase 2 are not responsible for PTX3 oxidation. PTX3 multimer deposition within the aorta is neutrophil dependent. Circulating proteome alterations, in both humans and mice, in response to endotoxin were mapped utilizing proteomic approaches. Multimeric pentraxin-3 (PTX3) plasma and aortic levels mimicked changes observed in proteins of known neutrophil origin in response to endotoxin. Genetic mouse models ruled out myeloperoxidase and NADPH oxidase 2 as drivers of PTX3 oxidation; however, neutrophil extravasation promotes PTX3 deposition within the vasculature. The results reveal a link between the systemic inflammatory response and the neutrophil-dependent vascular deposition of multimeric PTX3.