MicroRNA-18a Improves Human Cerebral Arteriovenous Malformation Endothelial Cell Function
MicroRNA-18a Improves Human Cerebral Arteriovenous Malformation Endothelial Cell Function
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DOI：
10.1161/strokeaha.113.003578
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发表时间：
2014-01-01
期刊：
影响因子：
8.3
通讯作者：
Hofman, Florence M.
中科院分区：
文献类型：
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作者：
Ferreira, Raquel;Santos, Tiago;Hofman, Florence M.
背景和目的脑动静脉畸形（AVM）是一种破坏正常血流并导致严重神经功能损害或死亡的血管疾病。AVM衍生的脑内皮细胞（AVM-BEC）的异常功能是疾病的标志。我们的目的是使用microRNA-18 a（miR-18 a）作为治疗剂，以改善AVM-BEC function.Methods人AVM-BEC的生长因子的生产和增殖进行了测试，在不同的剪切流条件下，并评估小管形成。血小板反应蛋白-1，抑制剂的DNA结合蛋白1，和血管内皮生长因子（VEGF）同种型mRNA水平进行定量实时聚合酶链反应。采用酶联免疫吸附试验测定血小板反应蛋白-1、VEGF-A和VEGF-D蛋白表达。增殖和小管形成进行了评估，分别使用溴脱氧尿苷掺入和生长因子减少Matrigel assays.Results miR-18 a增加血小板反应蛋白-1的生产，但减少DNA结合蛋白1，血小板反应蛋白-1的转录抑制剂的抑制剂。miR-18 a降低VEGF-A和VEGF-D水平，两者在未处理的AVM-BEC中均过表达。这是第一项报告AVM中VEGF-D过表达的研究。这些影响在动脉剪切流条件下最为突出。miR-18 a还能降低AVM-BEC增殖，促进小管形成，并在无外源性转染试剂的情况下被AVM-BEC有效内化。结论我们报道了VEGF-D在AVM中的过表达以及miR-18 a诱导AVM-BEC功能更正常的能力。这突出了microRNA作为AVM和其他血管疾病治疗的临床潜力。
Background and Purpose Cerebral arteriovenous malformation (AVM) is a vascular disease that disrupts normal blood flow and leads to serious neurological impairment or death. Aberrant functions of AVM-derived brain endothelial cells (AVM-BECs) are a disease hallmark. Our aim was to use microRNA-18a (miR-18a) as a therapeutic agent to improve AVM-BEC function.Methods Human AVM-BECs were tested for growth factor production and proliferation under different shear flow conditions and evaluated for tubule formation. Thrombospondin-1, inhibitor of DNA-binding protein 1, and vascular endothelial growth factor (VEGF) isotype mRNA levels were quantified by quantitative real-time polymerase chain reaction. Thrombospondin-1, VEGF-A, and VEGF-D protein expression was measured using enzyme-linked immunosorbent assay. Proliferation and tubule formation were evaluated using bromodeoxyuridine incorporation and growth factor-reduced Matrigel assays, respectively.Results miR-18a increased thrombospondin-1 production but decreased inhibitor of DNA-binding protein 1, a transcriptional repressor of thrombospondin-1. miR-18a reduced VEGF-A and VEGF-D levels, both overexpressed in untreated AVM-BECs. This is the first study reporting VEGF-D overexpression in AVM. These effects were most prominent under arterial shear flow conditions. miR-18a also reduced AVM-BEC proliferation, improved tubule formation, and was effectively internalized by AVM-BECs in the absence of extraneous transfection reagents.Conclusions We report VEGF-D overexpression in AVM and the capacity of miR-18a to induce AVM-BECs to function more normally. This highlights the clinical potential of microRNA as a treatment for AVM and other vascular diseases.