Identification of activators of ERK5 transcriptional activity by high-throughput screening and the role of endothelial ERK5 in vasoprotective effects induced by statins and antimalarial agents.
Identification of activators of ERK5 transcriptional activity by high-throughput screening and the role of endothelial ERK5 in vasoprotective effects induced by statins and antimalarial agents.
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DOI：
10.4049/jimmunol.1400571
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发表时间：
2014-10-01
期刊：
影响因子：
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通讯作者：
Abe J
中科院分区：
文献类型：
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作者：
Le NT;Takei Y;Izawa-Ishizawa Y;Heo KS;Lee H;Smrcka AV;Miller BL;Ko KA;Ture S;Morrell C;Fujiwara K;Akaike M;Abe J
由于细胞外信号调节激酶5（ERK 5）抑制内皮炎症和功能障碍，激活ERK 5可能是一种新的方法来保护血管内皮细胞（EC）免受各种血管病理条件。我们已经确定了通过ERK 5激活保护EC的小分子，并确定了它们对预防心脏移植排斥反应的贡献。使用高通量筛选（HTS），我们鉴定了某些他汀类药物和抗疟疾剂，包括氯喹（CQ）、羟氯喹（HCQ）和奎纳克林（QC）作为强ERK 5“激活剂”。匹伐他汀增强ERK 5的转录活性和Kruppel样因子-2（KLF 2）在培养的人和牛EC中的表达，但这些作用被ERK 5的耗竭所消除。CQ和HCQ上调ERK 5激酶活性，抑制VCAM-1的表达，ERK 5依赖，但MEK 5和KLF 2/4的非依赖性方式。白细胞滚动和血管反应性被用来评估体内内皮功能，我们发现EC特异性ERK 5敲除（ERK 5-EKO）小鼠表现出白细胞滚动增加和血管反应性受损，这不能被匹伐他汀纠正。还通过将从野生型（WT）或ERK 5-EKO小鼠获得的心脏异位移植到同种异体移植受体小鼠中来检查内皮ERK 5在急性心脏同种异体移植物排斥中的作用。观察到炎性基因表达和CD 45阳性细胞浸润到移植物中的强烈增加。这些组织排斥反应在WT心脏中被匹伐他汀抑制，但在ERK 5-EKO心脏中不被抑制。我们的研究已经确定了他汀类药物和抗疟疾药物作为强ERK 5激活剂，并表明ERK 5激活可预防内皮炎症和功能障碍以及急性同种异体移植排斥反应。
Because extracellular signal-regulated kinase 5 (ERK5) inhibits endothelial inflammation and dysfunction, activating ERK5 might be a novel approach to protecting vascular endothelial cells (ECs) against various pathological conditions of the blood vessel. We have identified small molecules that protect ECs via ERK5 activation and determined their contribution to preventing cardiac allograft rejection. Using high throughput screening (HTS), we identified certain statins and anti-malarial agents including chloroquine (CQ), hydroxychloroquine (HCQ), and quinacrine (QC) as strong ERK5 “activators”. Pitavastatin enhanced ERK5 transcriptional activity and Kruppel-like factor-2 (KLF2) expression in cultured human and bovine ECs, but these effects were abolished by the depletion of ERK5. CQ and HCQ up-regulated ERK5 kinase activity and inhibited VCAM-1 expression in an ERK5-dependent but MEK5- and KLF2/4-independent manner. Leukocyte rolling and vascular reactivity were used to evaluate endothelial function in vivo, and we found that EC-specific ERK5 knockout (ERK5-EKO) mice exhibited increased leukocyte rolling and impaired vascular reactivity, which could not be corrected by pitavastatin. The role of endothelial ERK5 in acute cardiac allograft rejection was also examined by heterotopic grafting of the heart obtained from either wild type (WT) or ERK5-EKO mice into allomismatched recipient mice. A robust increase in both inflammatory gene expression and CD45-positive cell infiltration into the graft was observed. These tissue rejection responses were inhibited by pitavastatin in WT but not ERK5-EKO hearts. Our study has identified statins and anti-malarial drugs as strong ERK5 activators and shown that ERK5 activation is preventive of endothelial inflammation and dysfunction and acute allograft rejection.