Initial sites of hepadnavirus integration into host genome in human hepatocytes and in the woodchuck model of hepatitis B-associated hepatocellular carcinoma.
Initial sites of hepadnavirus integration into host genome in human hepatocytes and in the woodchuck model of hepatitis B-associated hepatocellular carcinoma.
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在人肝细胞和肝炎B相关的肝细胞癌肝炎模型中，肝病病毒整合到宿主基因组中的初始部位。
DOI：
10.1038/oncsis.2017.22
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发表时间：
2017-04-17
期刊：
影响因子：
6.2
通讯作者：
Michalak TI
中科院分区：
文献类型：
--
作者：
Chauhan R;Churchill ND;Mulrooney-Cousins PM;Michalak TI
乙型肝炎病毒（HBV）和密切相关的土拨鼠肝炎病毒（WHV）是引发原发性肝细胞癌（HCC）发展的强致癌物。肝病毒-宿主基因组整合的初始位点、它们的多样性和形成动力学可能是病毒持续存在和HCC发生和进展的核心。为了认识早期病毒-宿主相互作用的性质，我们探索了人类肝细胞样HepaRG细胞与真实HBV和幼年土拨鼠与WHV的从头感染。在暴露于HBV后几分钟开始分析HepaRG，而土拨鼠在感染WHV后1或3小时和6周进行肝活检。利用逆转录PCR和克隆测序技术鉴定病毒与宿主的基因组连接。HBV和WHV DNA及其复制中间体在病毒暴露后1小时可检出。同时，检测到HBV DNA整合到各种宿主基因中。值得注意的是，HBV X基因与逆转录转座子序列（如LINE1和LINE2）的连接在感染后不久就变得突出。在土拨鼠中，whvx和preS序列在感染后1和3小时插入宿主基因组，证实了肝病毒在自然条件下入侵后很快整合到肝细胞DNA中。HBV和WHV X基因增强子II/核心启动子序列最常与宿主DNA形成初始连接。此外，1株hp - i出现了多个病毒DNA融合。在被感染的肝细胞和土拨鼠的肝脏中。总之，HBV DNA在感染后几乎立即与多种宿主序列整合，其中串联重复的非编码DNA是常见的。这项研究表明，HBV可以从感染开始就参与可移动的遗传元件，在整个宿主基因组中诱导促癌扰动。这种快速的病毒插入在土拨鼠的自然肝病毒感染中也很明显。
Hepatitis B virus (HBV) and the closely related woodchuck hepatitis virus (WHV) are potent carcinogens that trigger development of primary hepatocellular carcinoma (HCC). The initial sites of hepadnavirus–host genome integration, their diversity and kinetics of formation can be central to virus persistence and the initiation and progression of HCC. To recognize the nature of the very early virus–host interactions, we explored de novo infection of human hepatocyte-like HepaRG cells with authentic HBV and naive woodchucks with WHV. HepaRG were analyzed from several minutes post exposure to HBV onwards, whereas woodchuck liver biopsies at 1 or 3 h and 6 weeks post infection with WHV. Inverse PCR and clonal sequencing of the amplicons were applied to identify virus–host genomic junctions. HBV and WHV DNA and their replication intermediates became detectable in one hour after virus exposure. Concomitantly, HBV DNA integration into various host genes was detected. Notably, junctions of HBV X gene with retrotransposon sequences, such as LINE1 and LINE2, became prominent shortly after infection. In woodchucks, insertion of WHV X and preS sequences into host genome was evident at 1 and 3 h post infection (h.p.i.), confirming that hepadnavirus under natural conditions integrates into hepatocyte DNA soon after invasion. The HBV and WHV X gene enhancer II/core promotor sequence most often formed initial junctions with host DNA. Moreover, multiple virus–virus DNA fusions appeared from 1 h.p.i. onwards in both infected hepatocytes and woodchuck livers. In summary, HBV DNA integrates almost immediately after infection with a variety of host’s sequences, among which tandemly repeating non-coding DNAs are common. This study revealed that HBV can engage mobile genetic elements from the beginning of infection to induce pro-oncogenic perturbations throughout the host genome. Such swift virus insertion was also evident in natural hepadnaviral infection in woodchucks.
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影响因子：
50.3
作者：
Lau, Chi-Chiu;Sun, Tingting;Wong, Nathalie
通讯作者：
Wong, Nathalie
影响因子：
4.5
作者：
Ding D;Lou X;Hua D;Yu W;Li L;Wang J;Gao F;Zhao N;Ren G;Li L;Lin B
通讯作者：
Lin B
DOI：
10.1073/pnas.97.12.6634
发表时间：
2000-06-06
影响因子：
11.1
作者：
Bailey, JA;Carrel, L;Eichler, EE
通讯作者：
Eichler, EE
影响因子：
5.4
作者：
CHEN, IH;HUANG, CJ;TING, LP
通讯作者：
TING, LP
影响因子：
5.4
作者：
Gujar, Shashi A.;Mulrooney-Cousins, Patricia M.;Michalak, Tomasz I.
通讯作者：
Michalak, Tomasz I.