多重耐药的乙肝病毒在恩替卡韦抢救治疗克隆演变

StephenW

Clonal evolution of multidrug resistant hepatitis B virus during entecavir rescue therapy

    Young E. Chon1,2,†, Bora Jin3,†, Sang H. Ahn1,2,‡,*, Seungtaek Kim1,4,‡, Nam D. Kim5, Jeon H. Park6, Chung M. Nam7, Kyun-Hwan Kim8, Sun P. Hong9, Sung H. Choi3, Do Y. Kim1,2, Jun Y. Park1,2 andKwang-Hyub Han1,2

Article first published online: 13 MAY 2015

DOI: 10.1111/liv.12845

© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd

Issue
Liver International
Liver International

Volume 35, Issue 11, pages 2370–2383, November 2015


    1    Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea
    2    Liver Cirrhosis Clinical Research Center, Yonsei University Health System, Seoul, Korea
    3    Brain Korea 21 plus Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea
    4    Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Korea
    5    New Drug Development Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu, Korea
    6    Department of Microbiology and Immunology, Yonsei University College of Medicine, Seoul, Korea
    7    Department of Biostatistics, Yonsei University College of Medicine, Seoul, Korea
    8    Department of Pharmacology and Center for Cancer Research and Diagnostic Medicine, IBST, School of Medicine, Konkuk University, Seoul, Korea
    9    Research and Development Center, GeneMatrix Inc., Seongnam, Korea

    †    Young Eun Chon and Bora Jin have equally contributed to this work.
    ‡    Sang Hoon Ahn and Seungtaek Kim share the correspondence to this article.

* Correspondence
Sang Hoon Ahn, MD, PhD, Department of Internal Medicine, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun–gu, Seoul 120–752, Korea
Tel: 82 2 2228 1936
Fax: 82 2 393 6884
e-mail: [email protected]


Keywords:

    chronic hepatitis B;clonal analysis;colocation;drug resistance;entecavir;hepatitis B virus;multidrug resistance

Abstract
Background & Aims

Analysing the mutation pattern of multidrug resistance (MDR) is important in the treatment of chronic hepatitis B (CHB). In this study, the evolutionary pattern of MDR mutations was investigated in patients receiving entecavir (ETV) rescue therapy.
Methods

Eight CHB patients with lamivudine (LAM)- and adefovir (ADV)-resistant mutations showing suboptimal response to ETV and to subsequent ETV-plus-ADV therapy were enrolled. The clonal evolution of the mutation pattern was investigated through direct sequencing, multiplex restriction fragment mass polymorphism (RFMP), and clonal analysis and the utility of these methods was compared.
Results

Among 160 clones at baseline, wild-type hepatitis B virus (HBV) was present in 62 (38.8%), LAM-resistant mutations in 92 (57.6%) and ADV-resistant mutations in 55 (34.4%). LAM-resistant mutations increased to 70.6% at the end of ETV therapy and increased to 74.4% at the 12th month of ETV-plus-ADV therapy. During the same time periods, ETV-resistant mutations were present in 46.3% and 38.8%, and ADV-resistant mutations were present in 3.1% and 9.4% respectively. When 256 nucleotides from 32 samples were examined for mutations, clonal analysis detected 93 mutations (36.3%), direct sequencing detected 36 mutations (14.1%) and RFMP detected 73 mutations (28.5%). The sensitivity (73.1%, 95% CI; 64.1–82.1%) and specificity (96.9%, 95% CI; 94.4–99.4%) of RFMP were high, showing a concordance rate of 88.3% with the results from clonal analysis. All mutations exceeding 40% of the total clones detected by clonal analysis were also detected by RFMP.
Conclusions

The clonal evolution of the mutation pattern in MDR HBV showed the selection of LAM-resistant (±ETV-resistant) HBV during ETV rescue therapy, which may be the primary reason for patients' suboptimal response. Multiplex RFMP is a useful method for detecting MDR mutations in clinical practice.

StephenW

多重耐药的乙肝病毒在恩替卡韦抢救治疗克隆演变

    年轻E. Chon1,2，†，宝来Jin3，†，桑H. Ahn1,2，‡，*，Seungtaek Kim1,4，‡，南D. Kim5，全度妍H. Park6，钟M. Nam7，筠焕Kim8，孙P. Hong9，宋H. Choi3，难道Y. Kim1,2，君Y. Park1,2 andKwang，Hyub Han1,2

文章首次在网上公布：2015年5月13日

DOI：10.1111 / liv.12845

©2015年约翰·威利父子A / S。由John Wiley＆Sons出版有限公司

问题
肝国际
肝国际

第35卷，第11期，页2370至83年，2015年十一月


    内科，医学延世大学，首尔，韩国1系
    2肝硬化临床研究中心，延世大学医疗系统，韩国首尔
    3脑韩国21加项目的医学科学，医学延世大学，首尔，韩国
    4遣散生物医学科学研究所，医学延世大学，韩国首尔
    5新药开发中心，大邱庆北医创新基金，韩国大邱
    微生物学与免疫学，医学延世大学，首尔，韩国6系
    生物统计学7系，医学院，韩国首尔延世大学
    药理学和癌症研究中心和医学诊断，IBST，医学院，建国大学，韩国首尔的8部
    9研究与发展中心，GeneMatrix公司，城南，韩国

    †杨恩川川和宝来劲都平等地促进了这项工作。
    ‡相勋安与Seungtaek金分享与此相对应的文章。

*通讯
桑勋安，医学博士，内科，医学延世大学，延世大学50-RO，西大门区，首尔120-752，韩国部
联系电话：82 2 2228 1936年
传真：82 2 393 6884
电子邮件：[email protected]


关键词：

    慢性乙型肝炎;克隆分析;托管;耐药性;恩替卡韦;乙型肝炎病毒;多药耐药性

抽象的
背景和目的

浅析多药耐药（MDR）的突变模式是在慢性乙型肝炎（CHB）的治疗很重要。在这项研究中，多药耐药突变演化模式进行了研究在接受恩替卡韦（ETV）抢救治疗的患者。
方法

八慢性乙型肝炎患者，拉米夫定（LAM） - 和阿德福韦（ADV）耐药突变显示为ETV和后续ETV加ADV治疗反应不佳的患者。突变图案的克隆演变是通过直接测序研究，多重限制片段质量多态性（RFMP），并克隆分析及这些方法的效用进行比较。
结果

其中160个克隆的基础，野生型乙型肝炎病毒（HBV）是目前在62（38.8％），92（57.6％），LAM耐药突变，在55个（34.4％）ADV耐药突变。林耐药性突变增加至70.6％，在ETV治疗结束和增加至74.4％，在第12个月ETV加ADV疗法。在同一时间段，ETV耐药突变存在于46.3％和38.8％，而ADV耐药突变的3.1％和9.4％，分别出席了会议。当进行了检查的突变256个核苷酸，从32个样品，克隆分析检测到93突变（36.3％），直接测序检测36突变（14.1％）和RFMP检测73突变（28.5％）。灵敏度（73.1％，95％CI，64.1-82.1％）和特异性（96.9％，95％CI，94.4-99.4％）RFMP分别为高，显示出88.3％，与克隆分析结果的符合率。还通过RFMP检测到的所有突变超过由克隆分析所检测的总克隆的40％。
结论

在MDR HBV突变图案的克隆演变显示的LAM耐药（±ETV抗性）的HBV期间ETV抢救性治疗的选择，这可能是主要的原因，患者的不理想的反应。多路复用RFMP是用于检测耐药突变在临床实践中的有用方法。