한빛사논문
- 조회1,911
Eun-Sook Park1, a, Ah Ram Lee1, a, Doo Hyun Kim1, a, Jeong-Hoon Lee2, a, b, Jeong-Ju Yoo2, 3, Sung Hyun Ahn1, Heewoo Sim1, Soree Park1, Hong Seok Kang1, Juhee Won1, Yea Na Ha1, Gu-Choul Shin1, So Young Kwon4, Yong Kwang Park5, Byeong-Sun Choi5, Yun Bin Lee2, 6, Nakcheol Jeong7, Yohan An8, Young Seok Ju8, 9, Su Jong Yu2, Hee Bok Chae10, Kyung-Sang Yu11, Yoon Jun Kim2, Jung-Hwan Yoon2, Fabien Zoulim12, Kyun-Hwan Kim1, 13, 14, b
1 Department of Pharmacology and Center for Cancer Research and Diagnostic Medicine, IBST, School of Medicine, Konkuk University, Seoul, Korea
2 Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine, Seoul, Korea
3 Department of Gastroenterology and Hepatology, Soonchunhyang University Bucheon Hospital, Gyeonggido, Korea
4 Department of Internal Medicine, School of Medicine, Konkuk University, Seoul, Korea
5 Division of AIDS, Center for Immunology and Pathology, Korea National Institute of Health, Korea Center for Disease Control and Prevention, Osong, Chungbuk, Korea
6 Department of Internal Medicine, CHA Bundang Medical Center, CHA University, Seongnam, Gyeonggido, Korea
7 Department of Chemistry, Korea University, Seoul, Korea
8 Biomedical Science and Engineering Interdisciplinary Program, Korea Advanced Institute of Science and Technology, Daejon, Korea
9 Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejon, Korea
10 Department of Internal Medicine, Chungbuk National University College of Medicine, Cheongju, Korea
11 Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine, Seoul, Korea
12 INSERM Unite 1052, Cancer Research Center of Lyon, Hospices Civils de Lyon, Lyon University, Lyon, France
13 KU Open Innovation Center, Konkuk University, Seoul, Korea
14 Research Institute of Medical Sciences, Konkuk University, Seoul, Korea
aThese four authors are co-first authors
bThese two authors are co-corresponding authors
*Corresponding authors.
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.
Keywords : capsid assembly modulator; entecavir; nucleotide analogue; CYEI; rtS106C; rtH126Y; rtD134E; rtL269I
논문정보
- Research article
- 2019년 02월 (BRIC 등록일 2019-02-28)
- 국내(교신)+국외 연구진
- 의약학 > 응용의학
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