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

IL-35抑制肠道微生物区系产生的尿毒症毒素加速的内皮细胞 激活 慢性肾脏疾病(CKD)影响着全球15%的成年人口。CKD推广 心血管发病率和死亡率。内皮细胞(EC)激活/功能障碍增加 而血管炎症在加速CKD的发展中起着关键作用 心血管疾病(CVD)，开始于CKD的早期阶段。新的疗法是 迫切需要抑制CKD加速的EC激活/功能障碍。EC功能障碍是 与一氧化氮(NO)的减少有关，而我们的新JBC论文显示EC 激活特征包括：1)细胞因子和趋化因子的分泌增加；2)上调 EC黏附分子；3)上调额外的DAMP受体；以及4)上调 T细胞共刺激/共抑制受体。旨在减少EC的新战略 活化/功能障碍和血管炎症可能在早期提供有效的治疗 慢性肾脏病分期。三甲胺-N-氧化物(TMAO)是一种肠道微生物区系，由胆碱产生 代谢物。TMAO是一种新发现的尿毒症毒素，在CKD和CKD中强烈升高。 与动脉粥样硬化性心血管疾病相关。TMAO诱导内皮细胞功能障碍和血管增加 炎症通过与G蛋白偶联受体结合，从而激活丝裂原激活 蛋白激酶和核因子-κB，增加循环细胞因子。我们和其他人报道说 白介素35(IL-35)是一种新的、强大的抗炎细胞因子，可抑制多种 发炎。CKD患者血浆IL-35水平升高。因此，中央 该方案的假设是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 这一假说通过以下三个目的：1)确定表达和抑制功能 TMAO诱导人和小鼠主动脉内皮细胞IL-35/IL-35R亚基的表达 (2)确定IL-35抑制CKD小鼠生长的分子机制 通过抑制线粒体活性氧(MtROS)的产生来激活EC， 抑制caspase-1(CASP1)典范/casp11非典范炎症小体信号，以及 抑制组蛋白3赖氨酸14乙酰化(H3K14ac)诱导EC激活基因表达； 3)确定两个IL-35亚基(p35和EBI3)和一个IL-35的抑制作用 慢性肾脏病小鼠模型中受体(IL-35R)亚单位(IL-12Rβ-2)的表达。《目标3》将通过 一种新的IL-35治疗方法(功能获得)在CKD小鼠模型中的应用，以及四种 功能性CKD模型包括p35-/-CKD小鼠、EBI-3-/-CKD小鼠、全局IL-12Rβ2-/-CKD小鼠 小鼠和EC特异性IL12Rb2-/-CKD小鼠确定IL-35的抑制作用 慢性肾脏病中内皮细胞功能障碍和血管炎症的信号转导。这件事的意义 建议是，这些研究的成功应该在实地产生重大影响，以及 提供了对IL-35抑制TMAO诱导的EC激活和 CKD加速血管炎症，这可能导致新的潜在开发 治疗CKD加速的心血管疾病。

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.