Enterovirus Persistence in Cardiac Cells of Patients With Idiopathic Dilated Cardiomyopathy Is Linked to 5' Terminal Genomic RNA-Deleted Viral Populations With Viral-Encoded Proteinase Activities
Enterovirus Persistence in Cardiac Cells of Patients With Idiopathic Dilated Cardiomyopathy Is Linked to 5' Terminal Genomic RNA-Deleted Viral Populations With Viral-Encoded Proteinase Activities
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DOI：
10.1161/circulationaha.118.035966
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发表时间：
2019-05-14
期刊：
影响因子：
37.8
通讯作者：
Andreoletti, Laurent
中科院分区：
文献类型：
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作者：
Bouin, Alexis;Gretteau, Paul-Antoine;Andreoletti, Laurent
背景技术背景：B组肠道病毒是急性心肌炎的常见原因，急性心肌炎可能是慢性心肌炎和扩张型心肌病的前兆，是心脏移植的主要原因。方法：从扩张型心肌病患者心肌组织中提取总RNA，对肠道病毒RNA的5'端序列进行鉴定。在下一代RNA测序后，在体外产生了模拟患者组织中发现的肠道病毒RNA序列的病毒cDNA克隆，并在培养的原代人心肌细胞上评估了它们的复制和对宿主细胞功能的影响。主要肠道病毒B群体的特征是5'末端基因组RNA缺失范围为17至50个核苷酸，这些群体单独或与低比例的肠道病毒相关。完整的5 ′基因组末端。原位杂交和免疫组化检测这些持久的基因组在心肌细胞簇。将病毒RNA转染到原代人心肌细胞中表明，缺失形式的基因组RNA在没有可检测到的病毒空斑形成的情况下显示早期复制活性，而混合的缺失和完整形式产生的颗粒能够诱导细胞病变效应，其水平与单独使用全长形式所观察到的水平不同。此外，缺失或全长和混合形式的病毒RNA能够指导翻译和生产的蛋白水解活性病毒蛋白酶2A在人类心肌细胞。我们证明，持续存在的病毒形式是由B型肠道病毒在其基因组RNA中携带一个5'末端缺失组成的，这些病毒单独或与完整的在原因不明的扩张型心肌病病例中，辅助RNA的长度群体可通过病毒蛋白酶2A的蛋白水解活性损害心肌细胞功能。这些结果提供了一个更好的理解的基础EV形式在人类心脏组织中的持久性的分子机制，并应刺激发展的新的治疗策略的基础上的特异性抑制剂的柯萨奇病毒B蛋白酶2A活性的急性和慢性心脏感染。
BACKGROUND: Group B enteroviruses are common causes of acute myocarditis, which can be a precursor of chronic myocarditis and dilated cardiomyopathy, leading causes of heart transplantation. To date, the specific viral functions involved in the development of dilated cardiomyopathy remain unclear.METHODS: Total RNA from cardiac tissue of patients with dilated cardiomyopathy was extracted, and sequences corresponding to the 5' termini of enterovirus RNAs were identified. After next-generation RNA sequencing, viral cDNA clones mimicking the enterovirus RNA sequences found in patient tissues were generated in vitro, and their replication and impact on host cell functions were assessed on primary human cardiac cells in culture.RESULTS: Major enterovirus B populations characterized by 5' terminal genomic RNA deletions ranging from 17 to 50 nucleotides were identified either alone or associated with low proportions of intact 5' genomic termini. In situ hybridization and immunohistological assays detected these persistent genomes in clusters of cardiomyocytes. Transfection of viral RNA into primary human cardiomyocytes demonstrated that deleted forms of genomic RNAs displayed early replication activities in the absence of detectable viral plaque formation, whereas mixed deleted and complete forms generated particles capable of inducing cytopathic effects at levels distinct from those observed with full-length forms alone. Moreover, deleted or full-length and mixed forms of viral RNA were capable of directing translation and production of proteolytically active viral proteinase 2A in human cardiomyocytes.CONCLUSIONS: We demonstrate that persistent viral forms are composed of B-type enteroviruses harboring a 5' terminal deletion in their genomic RNAs and that these viruses alone or associated with full-length populations of helper RNAs could impair cardiomyocyte functions by the proteolytic activity of viral proteinase 2A in cases of unexplained dilated cardiomyopathy. These results provide a better understanding of the molecular mechanisms that underlie the persistence of EV forms in human cardiac tissues and should stimulate the development of new therapeutic strategies based on specific inhibitors of the coxsackievirus B proteinase 2A activity for acute and chronic cardiac infections.