PMID- 33562603 OWN - NLM STAT- MEDLINE DCOM- 20210906 LR - 20210906 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 22 IP - 4 DP - 2021 Feb 5 TI - Distinctive HBV Replication Capacity and Susceptibility to Tenofovir Induced by a Polymerase Point Mutation in Hepatoma Cell Lines and Primary Human Hepatocytes. LID - 10.3390/ijms22041606 [doi] LID - 1606 AB - Tenofovir disoproxil fumarate (TDF) has been regarded as the most potent drug for treating patients with chronic hepatitis B (CHB). However recently, viral mutations associated with tenofovir have been reported. Here, we found a CHB patient with suboptimal response after more than 4 years of TDF treatment. Clonal analysis of hepatitis B virus (HBV) isolated from sequential sera of this patient identified the seven previously reported TDF-resistant mutations (CYELMVI). Interestingly, a threonine to alanine mutation at the 301 amino acid position of the reverse-transcriptase (RT) domain, (rtT301A), was commonly accompanied with CYELMVI at a high rate (72.7%). Since the rtT301A mutation has not been reported yet, we investigated the role of this naturally occurring mutation on the viral replication and susceptibility to tenofovir in various liver cells (hepatoma cells as well as primary human hepatocytes). A cell-based phenotypic assay revealed that the rtT301A mutation dramatically impaired the replication ability with meaningful reduction in sensitivity to tenofovir in hepatoma cell lines. However, attenuated viral replication by the rtT301A mutation was significantly restored in primary human hepatocytes (PHHs). Our findings suggest that the replication capability and drug sensitivity of HBV is different between hepatoma cell lines and PHHs. Therefore, our study emphasizes that validation studies should be performed not only in the liver cancer cell lines but also in the PHHs to understand the exact viral fitness under antiviral pressure in patients. FAU - Lee, Ah Ram AU - Lee AR AUID- ORCID: 0000-0001-6075-3352 AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Cho, Ju-Yeon AU - Cho JY AD - Department of Internal Medicine, College of Medicine, Chosun University, Gwangju 61452, Korea. FAU - Kim, Jong Chul AU - Kim JC AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Dezhbord, Mehrangiz AU - Dezhbord M AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Choo, Soo Yeun AU - Choo SY AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Ahn, Chang Hyun AU - Ahn CH AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Kim, Na Yeon AU - Kim NY AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Shin, Jae Jin AU - Shin JJ AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Park, Soree AU - Park S AUID- ORCID: 0000-0003-0500-2227 AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Park, Eun-Sook AU - Park ES AD - Department of Pharmacology, School of Medicine, Konkuk University, Seoul 05029, Korea. FAU - Won, Juhee AU - Won J AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. FAU - Kim, Dong-Sik AU - Kim DS AUID- ORCID: 0000-0002-0608-1580 AD - Department of Surgery, Division of HBP Surgery and Liver Transplantation, College of Medicine, Korea University, Seoul 02841, Korea. FAU - Lee, Jeong-Hoon AU - Lee JH AUID- ORCID: 0000-0002-0315-2080 AD - Department of Internal Medicine and Liver Research Institute, College of Medicine, Seoul National University, Seoul 03080, Korea. FAU - Kim, Kyun-Hwan AU - Kim KH AUID- ORCID: 0000-0001-5266-072X AD - Department of Precision Medicine, School of Medicine, Sungkyunkwan University, Suwon 16419, Korea. LA - eng PT - Case Reports PT - Journal Article DEP - 20210205 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - 0 (Antiviral Agents) RN - 0 (Reverse Transcriptase Inhibitors) RN - 0 (Viral Proteins) RN - 99YXE507IL (Tenofovir) RN - EC 2.7.7.49 (RNA-Directed DNA Polymerase) SB - IM MH - Antiviral Agents/pharmacology MH - Carcinoma, Hepatocellular/genetics MH - Cell Line, Tumor MH - Cells, Cultured MH - Drug Resistance, Viral/genetics MH - Female MH - Genes, Viral MH - Hep G2 Cells MH - Hepatitis B virus/*drug effects/genetics/physiology MH - Hepatitis B, Chronic/drug therapy/genetics/virology MH - Hepatocytes/*drug effects/metabolism/*virology MH - Humans MH - Liver Neoplasms/genetics MH - Middle Aged MH - Point Mutation MH - RNA-Directed DNA Polymerase/genetics MH - Reverse Transcriptase Inhibitors/pharmacology MH - Tenofovir/*pharmacology MH - Viral Proteins/genetics MH - Virus Replication/drug effects/genetics PMC - PMC7914950 OTO - NOTNLM OT - drug resistance OT - hepatitis B virus OT - nucleos(t)ide analog OT - reverse transcription OT - tenofovir disoproxil fumarate (TDF) COIS- The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. EDAT- 2021/02/11 06:00 MHDA- 2021/09/07 06:00 PMCR- 2021/02/05 CRDT- 2021/02/10 01:02 PHST- 2020/12/17 00:00 [received] PHST- 2021/01/27 00:00 [revised] PHST- 2021/01/29 00:00 [accepted] PHST- 2021/02/10 01:02 [entrez] PHST- 2021/02/11 06:00 [pubmed] PHST- 2021/09/07 06:00 [medline] PHST- 2021/02/05 00:00 [pmc-release] AID - ijms22041606 [pii] AID - ijms-22-01606 [pii] AID - 10.3390/ijms22041606 [doi] PST - epublish SO - Int J Mol Sci. 2021 Feb 5;22(4):1606. doi: 10.3390/ijms22041606.