IL-35 inhibits gut microbiota-produced uremic toxin-accelerated endothelial cell activation

IL-35抑制肠道菌群产生的尿毒症毒素加速的内皮细胞 激活 慢性肾脏病（CKD）影响着全球15%的成年人口。CKD促进 心血管疾病发病率和死亡率。内皮细胞（EC）活化/功能障碍增加 和血管炎症在CKD的发展中起着至关重要的作用 心血管疾病（CVD），开始于CKD的早期阶段。新的治疗方法 迫切需要抑制CKD加速的EC活化/功能障碍。EC功能障碍是 与减少一氧化氮（NO）的产生有关，而我们新的JBC论文显示EC 活化特征包括：1）增加细胞因子和趋化因子的分泌; 2）上调 EC粘附分子的上调; 3）额外的DAMP受体的上调;和4）EC粘附分子的上调。 T细胞共刺激/共抑制受体。旨在减少EC的新策略 激活/功能障碍和血管炎症可能在早期提供有效的治疗。 CKD的阶段。三甲基胺-N-氧化物（TMAO）是一种肠道微生物群产生的胆碱衍生物， 代谢物。TMAO是一种新发现的尿毒症毒素，在CKD患者中强烈升高， 与动脉粥样硬化性心血管疾病有关。TMAO诱导EC功能障碍和增加的血管张力。 炎症通过结合G蛋白偶联受体，从而激活丝裂原活化 蛋白激酶和NF-κB，以及增加循环细胞因子。我们和其他人报告说， 白细胞介素-35（IL-35）是一种新的和强大的抗炎细胞因子，其抑制多种炎症反应， 炎症CKD患者血浆IL-35水平升高。因此中央 该建议假设IL-35抑制尿毒症毒素TMAO诱导的EC 活化/功能障碍和CKD加速的血管炎症。因此，我们将研究 该假说通过以下三个目的：1）确定表达和抑制功能 TMAO诱导的人和小鼠主动脉内皮细胞中IL-35/IL-35 R亚基的表达 2）确定IL-35抑制CKD小鼠MREC的分子机制， 通过抑制线粒体活性氧（mtROS）的产生激活EC， 抑制半胱天冬酶-1（caspase 1）典型/caspase 11非典型炎性体信号传导，和 抑制组蛋白3赖氨酸14乙酰化（H3 K14 ac）诱导的EC活化基因表达;以及 3)为了确定两种IL-35亚基（p35和EBI 3）和一种IL-35亚基（p35和EBI 3）的抑制作用， 受体（IL-35 R）亚单位（IL-12 R β2）。目标3将通过 在CKD小鼠模型中使用新的IL-35疗法（功能获得），以及四种IL-35丧失。 功能性CKD模型，包括p35-/- CKD小鼠、EBI-3-/- CKD小鼠、整体IL-12 R β2-/- CKD 小鼠和EC特异性IL 12 Rb 2-/- CKD小鼠，以确定IL-35的抑制作用 CKD中EC功能障碍和血管炎症的信号传导。其意义 建议是，这些研究的成功应该在该领域产生重大影响， 提供了IL-35抑制TMAO诱导的EC活化的新机制见解， CKD-加速血管炎症，这可能导致新的潜在发展， CKD加速的CVD的治疗。

IL-35 inhibits gut microbiota-produced uremic toxin-accelerated endothelial cell activation Chronic kidney disease (CKD) affects 15% of the adult population worldwide. CKD promotes cardiovascular morbidity and mortality. Increased endothelial cell (EC) activation/dysfunction and vascular inflammation play a critical role the development of CKD-accelerated cardiovascular disease (CVD), which begins in the early stages of CKD. New therapies are urgently needed to inhibit EC activation/dysfunction accelerated by CKD. EC dysfunction is associated with reduced nitric oxide (NO) production whereas our new JBC paper showed EC activation features include: 1) increased secretion of cytokines and chemokines; 2) upregulation of EC adhesion molecules; 3) upregulation of additional DAMP receptors; and 4) upregulation of T cell co-stimulation/co-inhibition receptors. Novel strategies targeted at reducing EC activation/dysfunction and vascular inflammation may provide effective treatment in the early stages of CKD. Trimethylamine-N-Oxide (TMAO) is a gut microbiota generated, choline-derived metabolite. TMAO is a newly identified as a uremic toxin, which is strongly elevated in CKD and associated with atherosclerotic CVDs. TMAO induces EC dysfunction and increased vascular inflammation via binding to G-protein coupled receptor, thereby activating mitogen‐activated protein kinase and NF‐κB, and increasing circulating cytokines. We and others reported that interleukin-35 (IL-35) is a new and powerful anti-inflammatory cytokine that inhibits various inflammation. Plasma IL-35 levels are increased in CKD patients. Therefore, the central hypothesis of this proposal is that IL-35 suppresses uremic toxin TMAO-induced EC activation/dysfunction and CKD-accelerated vascular inflammation. Therefore, we will examine this hypothesis via following three aims: 1) To determine expression and suppressive function of IL-35/IL-35R subunits in TMAO-induced human aortic ECs (HAECs) and mouse aortic ECs (MRECs) from CKD mice; 2) To determine the molecular mechanisms, by which IL-35 inhibits EC activation via inhibiting mitochondrial reactive oxygen species (mtROS) generation, suppressing caspase-1(casp1) canonical/casp11 non-canonical inflammasome signaling, and inhibiting histone 3 lysine 14 acetylation (H3K14ac) induced EC activation gene expression; and 3) To determine the suppressing roles of two IL-35 subunits (p35, and EBI3) and an IL-35 receptor (IL-35R) subunit (IL12Rβ2) in CKD mouse model. The aim 3 will be explored through the use of a novel IL-35 therapy (gain of function) in CKD mouse model, and four loss of function CKD models including p35-/- CKD mice, EBI-3-/- CKD mice, global IL-12Rβ2-/- CKD mice and EC-specific IL12Rb2-/- CKD mice to determine the suppressive roles of IL-35 signaling in EC dysfunction and vascular inflammation in CKD. The significance of this proposal is that the success of these studies should have a major impact in the field, and provide novel mechanistic insights into IL-35 in suppressing TMAO-induced EC activation and CKD-accelerated vascular inflammation, which could lead to the potential development of novel therapeutics for CKD-accelerated CVDs.