High expression of JC polyomavirus-encoded microRNAs in progressive multifocal leukoencephalopathy tissues and its repressive role in virus replication
High expression of JC polyomavirus-encoded microRNAs in progressive multifocal leukoencephalopathy tissues and its repressive role in virus replication
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DOI：
10.1371/journal.ppat.1008523
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发表时间：
2020-04-01
期刊：
影响因子：
6.7
通讯作者：
Katano, Harutaka
中科院分区：
文献类型：
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作者：
Takahashi, Kenta;Sato, Yuko;Katano, Harutaka
JC多瘤病毒(JCPyV，JCV)在免疫低下的宿主中引起进行性多灶性白质脑病(PML)。JCPyV在PML患者脑组织内的少突胶质细胞中复制。JCPyV基因组在编码大T抗原的区域编码一个微小RNA(MiRNA)。已在PML患者的组织和脑脊液样本中检测到JCPyV编码的miRNA(miR-J1)，但尚未有报道描述多瘤病毒编码的miRNA在病毒相关疾病患者的组织标本中的定位。在本研究中，我们通过原位杂交检测到JCPyV感染的PML组织样本中miR-J1在细胞核中的高表达。此外，原位杂交还发现BK多瘤病毒(BKPyV)编码的miRNA在BKPyV相关性肾病的皮损中表达。免疫组织化学检测JCPyV阳性的PML组织中，24/25(96%)有miR-J1-5p和-3p的原位杂交阳性信号。实时定量逆转录聚合酶链式反应检测到PML组织中miR-J1基因的拷贝数高于非PML组织中的拷贝数。下一代测序表明，与另一种成熟miRNA miR-J1-3p相比，miR-J1-5p是初级miRNA的成熟miRNA。重组JCPyV中miR-J1的缺失或突变促进了JCPyV DNA转染细胞中JCPyV编码蛋白的产生，提示多瘤病毒编码的miRNA可能对PML组织中的病毒复制具有抑制作用。病毒miRNA的原位杂交可能是诊断PML的有用工具。作者概述JCPyV可导致免疫受损宿主的PML。在PML患者中，JCPyV在脑组织的少突胶质细胞中复制。实时定量聚合酶链式反应已在PML患者的脑组织和脑脊液中检测到JCPyV编码的miRNA miR-J1，但尚未有报道描述多瘤病毒编码的miRNA在病毒相关疾病患者的组织标本中的定位。在本研究中，原位杂交清楚地表明miR-J1在PML组织样本中JCPyV感染细胞的细胞核中高表达。在BKPyV相关性肾病中也发现了高水平的BKPyV编码的miRNA表达。重组JCPyV的实验结果表明，miR-J1缺陷促进了JCPyV DNA转染细胞中JCPyV的复制。病毒miRNA的高表达对病毒复制具有抑制作用，提示病毒复制具有自身调节机制，病毒miRNA有可能成为治疗PML的新工具。
JC polyomavirus (JCPyV, JCV) causes progressive multifocal leukoencephalopathy (PML) in immunocompromised hosts. JCPyV replicates in oligodendrocytes within the brain tissue of patients with PML. The JCPyV genome encodes a microRNA (miRNA) in the region encoding the large T antigen. JCPyV-encoded miRNA (miR-J1) has been detected in the tissue and cerebrospinal fluid samples of patients with PML; however, there are no reports describing the localization of polyomavirus-encoded miRNA in histological samples of patients with virus-associated diseases. In the present study, we detected high miR-J1 expression in the nuclei of JCPyV-infected cells in PML tissue samples via in situ hybridization. Additionally, in situ hybridization also revealed the expression of BK polyomavirus (BKPyV)-encoded miRNA in lesions of BKPyV -associated nephropathy. In situ hybridization for miR-J1-5p and -3p showed positive signals in 24/25 (96%) of PML tissues that were positive for JCPyV by immunohistochemistry. Higher copy numbers of miR-J1 were detected in PML tissues than in non-PML tissues by real-time reverse transcription PCR. Next generation sequencing showed that miR-J1-5p, a mature miRNA of primary miRNA, was predominant in the lesions compared with miR-J1-3p, another mature miRNA. Deletion or mutation of miR-J1 in recombinant JCPyV promoted the production of JCPyV-encoded proteins in cells transfected with JCPyV DNA, suggesting that polyomavirus-encoded miRNA may have a repressive role in viral replication in PML tissues. In situ hybridization for viral miRNA may be a useful diagnostic tool for PML.Author summaryJCPyV causes PML in immunocompromised hosts. In patients with PML, JCPyV replicates in the oligodendrocytes of brain tissue. A JCPyV-encoded miRNA, miR-J1, has been detected by real-time PCR in the brain tissue and cerebrospinal fluid of patients with PML; however, there are no reports describing the localization of polyomavirus-encoded miRNA in histological samples of patients with virus-associated diseases. In this study, in situ hybridization clearly showed that miR-J1 was highly expressed in the nuclei of JCPyV-infected cells within PML tissue samples. A high level of BKPyV-encoded miRNA expression was also found in BKPyV-associated nephropathy. The results of experiments with recombinant JCPyV demonstrate that a defect of miR-J1 promotes JCPyV replication in cells transfected with JCPyV DNA. The observed high expression of viral miRNA, which has a repressive role in virus replication, suggests an autoregulation mechanism in virus replication as well as the possibility that viral miRNA may be a new therapeutic tool for PML.