Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells.
Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells.
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DOI：
10.1016/j.jhepr.2022.100534
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发表时间：
2022-09
期刊：
影响因子：
8.3
通讯作者：
Ploss, Alexander
中科院分区：
文献类型：
--
作者：
Wei, Lei;Cafiero, Thomas R.;Tseng, Anna;Gertje, Hans P.;Berneshawi, Andrew;Crossland, Nicholas A.;Ploss, Alexander
HBV的宿主范围很窄，人类和黑猩猩是唯一已知的自然宿主。在生物医学研究中常用的物种（如小鼠）中，耐药性的分子相关性目前还不完全清楚。人NTCP（hNTCP）在小鼠肝细胞中的表达使HBV能够进入，但随后在大多数鼠细胞中不会形成共价闭合环状（cccDNA）。尚不清楚这种cccDNA形成的阻断是否是由于松弛环状DNA（rcDNA）修复为cccDNA的缺陷。在这里，我们部署了体内和体外病毒学和生物化学方法来研究小鼠细胞是否含有一套完整的修复因子，能够将HBV rcDNA转化为cccDNA。我们证明了HBV cccDNA确实在小鼠细胞培养物中形成，或者当不含蛋白加合物的重组rcDNA直接引入细胞时在小鼠中形成。我们进一步表明，在人类细胞中将rcDNA修复为cccDNA所需的DNA滞后链合成中核心组分的小鼠直系同源物可以支持HBV生命周期中的这一关键步骤。值得注意的是，在我们的研究中使用了重组HBV rcDNA底物，无论是不含蛋白加合物还是含有中性抗生物素蛋白以模拟HBV聚合酶;目前尚不清楚小鼠和人细胞中HBV聚合酶的去除过程是否相同。总的来说，我们的数据表明，HBV生命周期在进入后被阻断，并且可能在小鼠细胞的修复阶段之前，这产生了关键的见解，这将有助于构建对HBV感染具有近交系易感性的小鼠模型。B型肝炎病毒（HBV）在自然界中只感染人类和黑猩猩。小鼠模型通常用于疾病发病机制建模和临床前研究，以评估干预措施的有效性和安全性，然后在人类参与者中进行测试。然而，由于小鼠对HBV感染不敏感，因此很难在小鼠模型中准确地模拟人类感染（并测试潜在的治疗方法）。在此，我们已经证明小鼠能够在HBV生命周期中执行关键步骤，收紧了HBV不能有效感染和复制小鼠的可能原因。小鼠RFC、POLD、LIG 1、PCNA、FEN 1可以催化cccDNA的形成。rcDNA到cccDNA的转化可以在体外和体内在小鼠细胞系中得到支持。rcDNA（+）和（-）链在体外可被小鼠和人宿主因子类似地修复。小鼠细胞中HBV生命周期的阻断可能发生在核内复制之前的步骤。
HBV has a narrow host restriction, with humans and chimpanzees representing the only known natural hosts. The molecular correlates of resistance in species that are commonly used in biomedical research, such as mice, are currently incompletely understood. Expression of human NTCP (hNTCP) in mouse hepatocytes enables HBV entry, but subsequently covalently closed circular (cccDNA) does not form in most murine cells. It is unknown if this blockade in cccDNA formation is due to deficiency in repair of relaxed circular DNA (rcDNA) to cccDNA. Here, we deployed both in vivo and in vitro virological and biochemical approaches to investigate if murine cells contain a complete set of repair factors capable of converting HBV rcDNA to cccDNA. We demonstrate that HBV cccDNA does form in murine cell culture or in mice when recombinant rcDNA without a protein adduct is directly introduced into cells. We further show that the murine orthologues of core components in DNA lagging strand synthesis, required for the repair of rcDNA to cccDNA in human cells, can support this crucial step in the HBV life cycle. It is worth noting that recombinant HBV rcDNA substrates, either without a protein adduct or containing neutravidin to mimic HBV polymerase, were used in our study; it remains unclear if the HBV polymerase removal processes are the same in mouse and human cells. Collectively, our data suggest that the HBV life cycle is blocked post entry and likely before the repair stage in mouse cells, which yields critical insights that will aid in the construction of a mouse model with inbred susceptibility to HBV infection. Hepatitis B virus (HBV) is only known to infect humans and chimpanzees in nature. Mouse models are often used in modeling disease pathogenesis and preclinical research to assess the efficacy and safety of interventions before they are then tested in human participants. However, because mice are not susceptible to HBV infection it is difficult to accurately model human infection (and test potential treatments) in mouse models. Herein, we have shown that mice are able to perform a key step in the HBV life cycle, tightening the net around the possible reason why HBV can not efficiently infect and replicate in mice. Mouse RFC, POLD, LIG1, PCNA, FEN1 can catalyze formation of cccDNA. rcDNA to cccDNA conversion can be supported in mouse cell lines in vitro and in vivo. rcDNA (+) and (-) strands can be repaired similarly by mouse and human host factors in vitro. Block in the HBV life-cycle in mouse cells is likely at (a) step(s) prior to intranuclear replication.
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影响因子：
6.7
作者：
Bility MT;Cheng L;Zhang Z;Luan Y;Li F;Chi L;Zhang L;Tu Z;Gao Y;Fu Y;Niu J;Wang F;Su L
通讯作者：
Su L
影响因子：
5.4
作者：
GUIDOTTI, LG;MATZKE, B;CHISARI, FV
通讯作者：
CHISARI, FV
DOI：
10.3390/v13050777
发表时间：
2021-04-28
期刊：
Viruses
影响因子：
--
作者：
Liu Y;Maya S;Ploss A
通讯作者：
Ploss A
影响因子：
5.4
作者：
Dion, Sarah;Bourgine, Maryline;Michel, Marie-Louise
通讯作者：
Michel, Marie-Louise
影响因子：
25.7
作者：
Billerbeck E;Mommersteeg MC;Shlomai A;Xiao JW;Andrus L;Bhatta A;Vercauteren K;Michailidis E;Dorner M;Krishnan A;Charlton MR;Chiriboga L;Rice CM;de Jong YP
通讯作者：
de Jong YP