靶向HBx-DDB1相互作用抑制硝唑尼特对cccDNA的HBV转录。

StephenW

Cell Mol Gastroenterol Hepatol. 2018 Oct 24;7(2):297-312. doi: 10.1016/j.jcmgh.2018.10.010. [Epub ahead of print]
Inhibition of HBV Transcription From cccDNA With Nitazoxanide by Targeting the HBx-DDB1 Interaction.
Sekiba K1, Otsuka M2, Ohno M1, Yamagami M1, Kishikawa T1, Suzuki T1, Ishibashi R1, Seimiya T1, Tanaka E1, Koike K1.
Author information

1
    Department of Gastroenterology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan.
2
    Department of Gastroenterology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan. Electronic address: [email protected].

Abstract
BACKGROUND & AIMS:

Hepatitis B virus (HBV) infection is a major health concern worldwide. Although currently used nucleos(t)ide analogs efficiently inhibit viral replication, viral proteins transcribed from the episomal viral covalently closed circular DNA (cccDNA) minichromosome continue to be expressed long-term. Because high viral RNA or antigen loads may play a biological role during this chronicity, the elimination of viral products is an ultimate goal of HBV treatment. HBV regulatory protein X (HBx) was recently found to promote transcription of cccDNA with degradation of Smc5/6 through the interaction of HBx with the host protein DDB1. Here, this protein-protein interaction was considered as a new molecular target of HBV treatment.
METHODS:

To identify candidate compounds that target the HBx-DDB1 interaction, a newly constructed split luciferase assay system was applied to comprehensive compound screening. The effects of the identified compounds on HBV transcription and cccDNA maintenance were determined using HBV minicircle DNA, which mimics HBV cccDNA, and the natural HBV infection model of human primary hepatocytes.
RESULTS:

We show that nitazoxanide (NTZ), a thiazolide anti-infective agent that has been approved by the FDA for protozoan enteritis, efficiently inhibits the HBx-DDB1 protein interaction. NTZ significantly restores Smc5 protein levels and suppresses viral transcription and viral protein production in the HBV minicircle system and in human primary hepatocytes naturally infected with HBV.
CONCLUSIONS:

These results indicate that NTZ, which targets an HBV-related viral-host protein interaction, may be a promising new therapeutic agent and a step toward a functional HBV cure.

Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.
KEYWORDS:

Drug Screening; Minicircle; Primary Hepatocyte Infection

PMID:
    30704981
DOI:
    10.1016/j.jcmgh.2018.10.010

StephenW

Cell Mol Gastroenterol Hepatol。 2018年10月24日; 7（2）：297-312。 doi：10.1016 / j.jcmgh.2018.10.010。 [印刷前的电子版]
靶向HBx-DDB1相互作用抑制硝唑尼特对cccDNA的HBV转录。
Sekiba K1，Otsuka M2，Ohno M1，Yamagami M1，Kishikawa T1，铃木T1，Ishibashi R1，Seimiya T1，Tanaka E1，Koike K1。
作者信息

1
    日本东京大学大学院医学研究科消化内科
2
    日本东京大学大学院医学研究科消化内科电子地址：[email protected]。

抽象
背景与目的：

乙型肝炎病毒（HBV）感染是全世界主要的健康问题。尽管目前使用的核苷（t）ide类似物有效地抑制病毒复制，但是从附加型病毒共价闭合环状DNA（cccDNA）微染色体转录的病毒蛋白质继续长期表达。因为高病毒RNA或抗原负荷可能在这种慢性期间发挥生物学作用，所以消除病毒产物是HBV治疗的最终目标。最近发现HBV调节蛋白X（HBx）通过HBx与宿主蛋白DDB1的相互作用促进cccDNA的转录，同时降解Smc5 / 6。在这里，这种蛋白质 - 蛋白质相互作用被认为是HBV治疗的新分子靶标。
方法：

为了鉴定靶向HBx-DDB1相互作用的候选化合物，将新构建的裂解荧光素酶测定系统应用于综合化合物筛选。使用模拟HBV cccDNA的HBV小环DNA和人原代肝细胞的天然HBV感染模型测定鉴定的化合物对HBV转录和cccDNA维持的影响。
结果：

我们发现硝唑尼特（NTZ）是一种噻唑化物抗感染剂，已被FDA批准用于原生动物肠炎，可有效抑制HBx-DDB1蛋白的相互作用。 NTZ显着恢复Smc5蛋白水平并抑制HBV小环系统和天然感染HBV的人原代肝细胞中的病毒转录和病毒蛋白产生。
结论：

这些结果表明，靶向HBV相关病毒 - 宿主蛋白相互作用的NTZ可能是一种有前景的新治疗剂，也是一种朝向功能性HBV治愈的步骤。

版权所有©2019作者。由Elsevier Inc.出版。保留所有权利。
关键词：

药物筛选;小环;原发性肝细胞感染

结论：
    30704981
DOI：
    10.1016 / j.jcmgh.2018.10.010

StephenW

https://www.cmghjournal.org/arti ... 18)30156-5/fulltext