Sodium Hydrogen Exchangers Contribute to Arenavirus Cell Entry
Sodium Hydrogen Exchangers Contribute to Arenavirus Cell Entry
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DOI：
10.1128/jvi.02110-13
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发表时间：
2014-01-01
影响因子：
5.4
通讯作者：
de la Torre, Juan C.
中科院分区：
文献类型：
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作者：
Iwasaki, Masaharu;Nhi Ngo;de la Torre, Juan C.
几种阿拉伯病毒，主要是拉萨病毒(LASV)，会在人类中引起出血热(HF)疾病，并在它们流行的地区造成巨大的公共卫生问题。此外，有证据表明，世界各地分布的原型性ArenaVirus淋巴细胞性脉络膜脑膜炎病毒(LCMV)是一种被忽视的人类病原体，具有临床意义。现有抗击人类致病ArenaVirus的设备有限，这凸显了开发新型抗ArenaVirus药物的重要性，这项任务将通过确定和确定有助于ArenaVirus生命周期的病毒-宿主细胞因素相互作用的特征来促进这项任务。全基因组小干扰RNA(SiRNA)筛选发现，钠氢交换器3(NHE3)是LCMV在HeLa细胞中有效增殖所必需的，但NHE活性对LCMV生活史的影响机制尚不清楚。在这里，我们显示了NHE抑制剂5-(N-乙基-N-异丙基)阿米洛利(EIPA)对啮齿动物(BHK-21)和人类(A549)细胞中LCMV增殖的强烈抑制。EIPA介导的抑制不是由于干扰病毒RNA复制、基因表达或发芽，而是由于阻止病毒细胞进入。EIPA还可抑制由猪瘟病毒LASV和Junin病毒(JUNV)的糖蛋白介导的细胞进入。药理学和遗传学研究表明，LCMV在A549细胞中的进入依赖于肌动蛋白重塑和Ak1，提示存在类似巨噬细胞吞噬作用的细胞进入途径。最后，正在探索的治疗心肌梗死的NHE抑制剂佐尼波利特抑制了LCMV在培养细胞中的传播。我们的发现表明，靶向NHEs可能是对抗人类致病Arena病毒的一种新策略。
Several arenaviruses, chiefly Lassa virus (LASV), cause hemorrhagic fever (HF) disease in humans and pose a great public health concern in the regions in which they are endemic. Moreover, evidence indicates that the worldwide-distributed prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) is a neglected human pathogen of clinical significance. The limited existing armamentarium to combat human-pathogenic arenaviruses underscores the importance of developing novel antiarenaviral drugs, a task that would be facilitated by the identification and characterization of virus-host cell factor interactions that contribute to the arenavirus life cycle. A genome-wide small interfering RNA (siRNA) screen identified sodium hydrogen exchanger 3 (NHE3) as required for efficient multiplication of LCMV in HeLa cells, but the mechanisms by which NHE activity contributed to the life cycle of LCMV remain unknown. Here we show that treatment with the NHE inhibitor 5-(N-ethyl-N-isopropyl) amiloride (EIPA) resulted in a robust inhibition of LCMV multiplication in both rodent (BHK-21) and human (A549) cells. EIPA-mediated inhibition was due not to interference with virus RNA replication, gene expression, or budding but rather to a blockade of virus cell entry. EIPA also inhibited cell entry mediated by the glycoproteins of the HF arenaviruses LASV and Junin virus (JUNV). Pharmacological and genetic studies revealed that cell entry of LCMV in A549 cells depended on actin remodeling and Pak1, suggesting a macropinocytosis-like cell entry pathway. Finally, zoniporide, an NHE inhibitor being explored as a therapeutic agent to treat myocardial infarction, inhibited LCMV propagation in culture cells. Our findings indicate that targeting NHEs could be a novel strategy to combat human-pathogenic arenaviruses.