Ischemic Cerebroprotection Conferred by Myeloid Lineage-Restricted or Global CD39 Transgene Expression.
Ischemic Cerebroprotection Conferred by Myeloid Lineage-Restricted or Global CD39 Transgene Expression.
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DOI：
10.1161/circulationaha.116.023301
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发表时间：
2017-06-13
期刊：
影响因子：
37.8
通讯作者：
Pinsky DJ
中科院分区：
文献类型：
--
作者：
Baek AE;Sutton NR;Petrovic-Djergovic D;Liao H;Ray JJ;Park J;Kanthi Y;Pinsky DJ
缺血事件后的脑组织损伤可因炎症和血栓形成而加剧。细胞外ATP和ADP水平升高与细胞损伤、炎症和血栓形成相关。外核苷三磷酸二磷酸水解酶-1（CD 39）是一种在白细胞和内皮细胞质膜上表达的酶，通过磷酸化ATP/ADP抑制血小板活化和白细胞浸润。为了研究增加的CD 39在体内脑缺血模型中的作用，我们开发了表达人CD 39（hCD 39）的转基因（TG）小鼠。在启动子和hCD 39转录起始位点之间插入终止序列，产生其中可以使用Cre重组酶小鼠组织特异性控制hCD 39表达的小鼠。我们产生的小鼠表达hCD 39全球或仅在骨髓系细胞。大脑中动脉（MCA）闭塞造成脑缺血。在48小时后通过MRI定量CT体积。与野生型（WT）小鼠相比，整体和TG hCD 39-和髓系（LysM）CD 39-过表达小鼠（TG n=9，LysM n=6）均表现出显著更小的脑梗死体积。通过流式细胞术分析缺血和对侧半球的白细胞。虽然对侧半球具有相等数量的巨噬细胞和中性粒细胞，但来自TG小鼠的缺血半球具有较少的浸润（n=4）。与WT小鼠（n=6）相比，TG小鼠显示出较少的神经功能缺损。这是第一个报告的转基因过表达的CD 39小鼠赋予保护性表型中风后，减少白细胞浸润，梗死体积较小，减少神经功能缺损。CD 39的过度表达，无论是整体还是在髓系细胞中，都能淬灭缺血后的白细胞隔离，并减少卒中诱导的神经损伤。
Cerebral tissue damage after an ischemic event can be exacerbated by inflammation and thrombosis. Elevated extracellular ATP and ADP levels are associated with cellular injury, inflammation and thrombosis. Ectonucleoside triphosphate diphosphohydrolase-1 (CD39), an enzyme expressed on the plasmalemma of leukocytes and endothelial cells, suppresses platelet activation and leukocyte infiltration by phosphohydrolyzing ATP/ADP. To investigate the effects of increased CD39 in an in vivo cerebral ischemia model, we developed a transgenic (TG) mouse expressing human CD39 (hCD39). A floxed-stop sequence was inserted between the promoter and the hCD39 transcriptional start site, generating a mouse in which the expression of hCD39 can be controlled tissue-specifically, using Cre recombinase mice. We generated mice that express hCD39 globally or in myeloid-lineage cells only. Cerebral ischemia was induced by middle cerebral artery (MCA) occlusion. Infarct volumes were quantified by MRI after 48 hours. Both global and TG hCD39- and myeloid lineage (LysM) CD39-overexpressing mice (TG n=9, LysM n=6) demonstrated significantly smaller cerebral infarct volumes compared to wild type (WT) mice. Leukocytes from ischemic and contralateral hemispheres were analyzed by flow cytometry. While contralateral hemispheres had equal numbers of macrophages and neutrophils, ischemic hemispheres from TG mice had less infiltration (n=4). TG mice showed less neurologic deficit compared to WT mice (n=6). This is the first report of transgenic overexpression of CD39 in mice imparting a protective phenotype following stroke, with reduced leukocyte infiltration, smaller infarct volumes, and decreased neurological deficit. CD39 overexpression, either globally or in myeloid lineage cells, quenches post-ischemic leukosequestration and reduces stroke-induced neurological injury.