富马酸替诺福韦二吡呋酯对耐药 HBV 克隆的影响。

Effect of tenofovir disoproxil fumarate on drug-resistant HBV clones.

机构信息

Department of Gastroenterology and Metabolism, Applied Life Science, Institute of Biomedical & Health Science, Hiroshima University, Hiroshima, Japan; Liver Research Project Center, Hiroshima University, Hiroshima, Japan; Natural Science Center for Basic Research and Development, Hiroshima University, Hiroshima, Japan.

出版信息

J Infect. 2016 Jan;72(1):91-102. doi: 10.1016/j.jinf.2015.09.038. Epub 2015 Oct 26.

DOI:10.1016/j.jinf.2015.09.038

PMID:26515673

Abstract

BACKGROUND & AIMS: Tenofovir disoproxil fumarate (TDF) has been approved for chronic hepatitis B treatment, and favorable susceptibility of hepatitis B virus (HBV) has been indicated. However, differences in TDF susceptibility among HBV genotypes and drug-resistant strains are unclear. In this study, TDF susceptibilities between genotypes A and C were evaluated in vitro and in vivo using several drug-resistant HBV clones.

METHODS

HBV expression plasmids were constructed from sera of HBV carriers, and drug-resistant substitutions were introduced by site-directed mutagenesis. TDF susceptibility was evaluated by changes of core-associated HBV replication intermediates in vitro or by change of serum HBV DNA in human hepatocyte chimeric mice carrying each HBV clone in vivo.

RESULTS

TDF susceptibilities of lamivudine-resistant clones (rtL180M/M204V) and lamivudine plus entecavir-resistant clones (rtL180M/S202G/M204V) were similar to wild type clones in vitro. However, lamivudine plus adefovir-resistant clones (rtA181T/N236T) acquired tolerance to TDF, and the rtN236T mutation was considered to be a causal substitution for TDF resistance. Furthermore, genotypic differences in TDF susceptibility were also observed between genotypes A and C in vitro, and the differences could be confirmed in vivo (p = 0.023).

CONCLUSIONS

The present study indicates that TDF susceptibility varies among HBV genotypes and drug-resistant HBV clones.

摘要

背景与目的

富马酸替诺福韦二吡呋酯（TDF）已被批准用于慢性乙型肝炎的治疗，且对乙型肝炎病毒（HBV）具有良好的敏感性。然而，不同基因型和耐药株的 HBV 对 TDF 的敏感性差异尚不清楚。本研究通过几种耐药 HBV 克隆，在体外和体内评估了基因型 A 和 C 之间 TDF 的敏感性。

方法

从 HBV 携带者的血清中构建 HBV 表达质粒，并通过定点诱变引入耐药性取代。通过改变核心相关 HBV 复制中间体的方式在体外或通过改变携带每种 HBV 克隆的人肝细胞嵌合体小鼠血清 HBV DNA 的方式在体内评估 TDF 的敏感性。

结果

拉米夫定耐药克隆（rtL180M/M204V）和拉米夫定加恩替卡韦耐药克隆（rtL180M/S202G/M204V）的 TDF 敏感性与野生型克隆相似。然而，拉米夫定加阿德福韦耐药克隆（rtA181T/N236T）对 TDF 产生了耐受性，且 rtN236T 突变被认为是导致 TDF 耐药的因果取代。此外，在体外还观察到基因型 A 和 C 之间 TDF 敏感性的基因型差异，且该差异可在体内得到证实（p=0.023）。