鉴定在慢性乙型肝炎患者中赋予替诺福韦耐药性的四重突变

StephenW

J Hepatol. 2019 Feb 19. pii: S0168-8278(19)30120-5. doi: 10.1016/j.jhep.2019.02.006. [Epub ahead of print]
Identification of a quadruple mutation that confers tenofovir resistance in chronic hepatitis B patients.
Park ES1, Lee AR2, Kim DH3, Lee JH4, Yoo JJ5, Ahn SH6, Sim H7, Park S8, Kang HS9, Won J10, Ha YN11, Shin GC12, Kwon SY13, Park YK14, Choi BS15, Lee YB16, Jeong N17, An Y18, Ju YS19, Yu SJ20, Chae HB21, Yu KS22, Kim YJ23, Yoon JH24, Zoulim F25, Kim KH26.
Author information
Abstract
BACKGROUND & AIMS:

Tenofovir disoproxil fumarate (TDF) is one the most potent nucleot(s)ide analogues for treating chronic hepatitis B virus (HBV) infection. Phenotypic resistance caused by genotypic resistance to TDF has not been reported. This study aimed to characterize HBV mutations that confer tenofovir resistance.
METHODS:

Two patients with viral breakthrough during treatment with TDF-containing regimens were prospectively enrolled. The gene encoding HBV reverse transcriptase (RT) was sequenced. Eleven HBV clones harboring a series of mutations in the RT gene were constructed by site-directed mutagenesis. Drug susceptibility of each clone was determined by Southern blot analysis and real-time PCR. Relative frequency of mutants was evaluated by ultra-deep sequencing and clonal analysis.
RESULTS:

Five mutations (rtS106C [C], rtH126Y [Y], rtD134E [E], rtM204I/V, and rtL269I [I]) were commonly found in viral isolates from two patients. The mutations C, Y, and E were novel mutations associated with drug resistance. In assays for drug susceptibility, the IC50 value for wild-type HBV was 3.8±0.6 µM, whereas the IC50 values for CYE and CYEI mutants were 14.1±1.8 and 58.1±0.9 µM, respectively. The IC90 value for wild-type HBV was 30±0.5 µM, whereas the IC90 values for CYE and CYEI mutants were 185±0.5 and 790±0.2 µM, respectively. All tenofovir-resistant mutants had similar susceptibility to a capsid assembly modulator, NVR 3-778 (IC50 < 0.4 µM), compared with wild-type (IC50 = 0.4 µM).
CONCLUSIONS:

Our study reveals that the quadruple (CYEI) mutation increases the amount of tenofovir required to inhibit HBV by 15.3-fold in IC50 and 26.3-fold in IC90. These results demonstrate that tenofovir-resistant HBV mutants can emerge, although the genetic barrier is high.
LAY SUMMARY:

Tenofovir is the most potent nucleoside analogue for the treatment of chronic hepatitis B virus infection and there has been no hepatitis B virus mutation that confers > 10-fold resistance to tenofovir up to 8 years. Herein, we firstly identified a quadruple mutation that confer 15.3-fold (IC50) and 26.3-fold (IC90) resistance to tenofovir from two patients who experienced viral breakthrough during tenofovir treatment.

Copyright © 2019. Published by Elsevier B.V.
KEYWORDS:

CYEI; capsid assembly modulator; entecavir; nucleotide analogue; rtD134E; rtH126Y; rtL269I; rtS106C

PMID:
    30794889
DOI:
    10.1016/j.jhep.2019.02.006

StephenW

J Hepatol。 2019年2月19日.pii：S0168-8278（19）30120-5。 doi：10.1016 / j.jhep.2019.02.006。 [印刷前的电子版]
鉴定在慢性乙型肝炎患者中赋予替诺福韦耐药性的四重突变。
Park ES1，Lee AR2，Kim DH3，Lee JH4，Yoo JJ5，Ahn SH6，Sim H7，Park S8，Kang HS9，Won J10，Ha YN11，Shin GC12，Kwon SY13，Park YK14，Choi BS15，Lee YB16，Jeong N17 ，Y Y18，Ju YS19，Yu SJ20，Chae HB21，Yu KS22，Kim YJ23，Yoon JH24，Zoulim F25，Kim KH26。
作者信息
抽象
背景与目的：

替诺福韦地索普西富马酸盐（TDF）是治疗慢性乙型肝炎病毒（HBV）感染的最有效的核苷酸类似物之一。尚未报道由对TDF的基因型抗性引起的表型抗性。本研究旨在表征赋予替诺福韦耐药性的HBV突变。
方法：

前瞻性研究了两例在含TDF治疗方案治疗期间病毒突破的患者。对编码HBV逆转录酶（RT）的基因进行测序。通过定点诱变构建了11个在RT基因中具有一系列突变的HBV克隆。通过Southern印迹分析和实时PCR确定每个克隆的药物敏感性。通过超深度测序和克隆分析评估突变体的相对频率。
结果：

在两名患者的病毒分离株中常见5种突变（rtS106C [C]，rtH126Y [Y]，rtD134E [E]，rtM204I / V和rtL269I [I]）。突变C，Y和E是与抗药性相关的新突变。在药物敏感性测定中，野生型HBV的IC50值为3.8±0.6μM，而CYE和CYEI突变体的IC50值分别为14.1±1.8和58.1±0.9μM。野生型HBV的IC90值为30±0.5μM，而CYE和CYEI突变体的IC90值分别为185±0.5和790±0.2μM。与野生型（IC50 =0.4μM）相比，所有替诺福韦抗性突变体对衣壳组装调节剂NVR 3-778（IC50 <0.4μM）具有相似的易感性。
结论：

我们的研究表明，四重（CYEI）突变使ICV抑制HBV所需的替诺福韦量增加15.3倍，IC90增加26.3倍。这些结果表明，尽管遗传屏障很高，但可以出现替诺福韦抗性HBV突变体。
LAY总结：

替诺福韦是用于治疗慢性乙型肝炎病毒感染的最有效的核苷类似物，并且没有乙型肝炎病毒突变赋予替诺福韦长达8年的> 10倍的抗性。在这里，我们首先发现了一种四重突变，它对来自在替诺福韦治疗期间经历病毒突破的两名患者的替诺福韦赋予15.3倍（IC50）和26.3倍（IC90）抗性。

版权所有©2019。Elsevier B.V.
关键词：

CYEI;衣壳组装调节器;恩替卡韦;核苷酸类似物; rtD134E; rtH126Y; rtL269I; rtS106C

结论：
    30794889
DOI：
    10.1016 / j.jhep.2019.02.006

默然10

NVR 3-778 也就是替诺耐药后就靠这个了呗

newchinabok

默然10 发表于 2019-2-26 20:01
NVR 3-778 也就是替诺耐药后就靠这个了呗

nvr3—778挂了，不强效，最快的是abi—h0731

默然10

abi—h0731
不是还没出来吗？出来估计也不一定吃的起。