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  • Komiyama, Yasuyuki  (5)
  • Maekawa, Shinya  (5)
  • 1
    Online Resource
    Online Resource
    Analysis of direct‐acting antiviral‐resistant hepatitis C virus haplotype diversity by single‐molecule and long‐read sequencing
    Ovid Technologies (Wolters Kluwer Health) ; 2022
    In:  Hepatology Communications Vol. 6, No. 7 ( 2022-07), p. 1634-1651
    Details
    In: Hepatology Communications, Ovid Technologies (Wolters Kluwer Health), Vol. 6, No. 7 ( 2022-07), p. 1634-1651
    Abstract: The method of analyzing individual resistant hepatitis C virus (HCV) by a combination of haplotyping and resistance‐associated substitution (RAS) has not been fully elucidated because conventional sequencing has only yielded short and fragmented viral genomes. We performed haplotype analysis of HCV mutations in 12 asunaprevir/daclatasvir treatment‐failure cases using the Oxford Nanopore sequencer. This enabled single‐molecule long‐read sequencing using rolling circle amplification (RCA) for correction of the sequencing error. RCA of the circularized reverse‐transcription polymerase chain reaction products successfully produced DNA longer than 30 kilobase pairs (kb) containing multiple tandem repeats of a target 3 kb HCV genome. The long‐read sequencing of these RCA products could determine the original sequence of the target single molecule as the consensus nucleotide sequence of the tandem repeats and revealed the presence of multiple viral haplotypes with the combination of various mutations in each host. In addition to already known signature RASs, such as NS3‐D168 and NS5A‐L31/Y93, there were various RASs specific to a different haplotype after treatment failure. The distribution of viral haplotype changed over time; some haplotypes disappeared without acquiring resistant mutations, and other haplotypes, which were not observed before treatment, appeared after treatment. Conclusion : The combination of various mutations other than the known signature RAS was suggested to influence the kinetics of individual HCV quasispecies in the direct‐acting antiviral treatment. HCV haplotype dynamic analysis will provide novel information on the role of HCV diversity within the host, which will be useful for elucidating the pathological mechanism of HCV‐related diseases.
    Type of Medium: Online Resource
    ISSN: 2471-254X , 2471-254X
    URL: Article
    DOI: 10.1002/hep4.1929
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2022
    detail.hit.zdb_id: 2881134-3
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  • 2
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    Significance of serum IP‐10/CXCL10 measurement in predicting post‐direct acting antiviral treatment liver function in patients with HCV‐decompensated liver cirrhosis
    Wiley ; 2023
    In:  Hepatology Research Vol. 53, No. 4 ( 2023-04), p. 280-288
    Details
    In: Hepatology Research, Wiley, Vol. 53, No. 4 ( 2023-04), p. 280-288
    Abstract: Recently, with the advent of sofosbuvir/velpatasvir therapy, sustained virological response (SVR) can now be achieved even in patients with decompensated cirrhosis (dLC). However, the prognosis after SVR does not always improve in dLC, and appropriate indicators enabling prediction of prognosis is desired. Patients and methods Serum IP‐10/CXCL10 levels were measured in 47 patients (15 chronic hepatitis [CH], 17 compensated cirrhosis [cLC] , and 15 dLC) receiving direct acting antiviral (DAA) therapy, and their changes during the therapy were examined. Results All the patients achieved SVR. In patients with CH, the average IP‐10 level was 367, 102, and 68 pg/ml respectively at baseline, at the end of therapy and at 12 weeks after SVR (SVR12), and was decreased upon DAA therapy ( P   〈  0.001). In patients with cLC, IP‐10 was respectively 215, 91, and 77 pg/ml, and was decreased upon DAA therapy ( P   〈  0.001) while it was 283, 131, and 182 pg/ml in patients with dLC and there was no evident decrease ( P  = 0.55). When patients with dLC were further classified depending on the difference in Child–Pugh (CP) score improvement at SVR12, a significant decrease in IP‐10 was observed after treatment in those with improvement ( P  = 0.023) while a significant increase was observed in those without improvement ( P  = 0.016). Conclusion While serum IP‐10 level was decreased in patients with CH/cLC and dLC with post‐SVR‐CP improvement following SVR, it was increased in patients with dLC without post‐SVR CP improvement. The result indicates that IP‐10 dynamics may be useful for predicting liver function after DAA therapy.
    Type of Medium: Online Resource
    ISSN: 1386-6346 , 1872-034X
    URL: Issue
    URL: Article
    DOI: 10.1111/hepr.v53.4
    DOI: 10.1111/hepr.13861
    Language: English
    Publisher: Wiley
    Publication Date: 2023
    detail.hit.zdb_id: 2006439-1
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  • 3
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    Deep sequencing analysis of serum hepatitis B virus‐RNA during nucleot(s)ide analogue therapy
    Wiley ; 2021
    In:  Hepatology Research Vol. 51, No. 1 ( 2021-01), p. 39-50
    Details
    In: Hepatology Research, Wiley, Vol. 51, No. 1 ( 2021-01), p. 39-50
    Abstract: Recently, serum hepatitis B virus (HBV)‐RNA has been reported to be detectable even when HBV particle production is inhibited by nucleot(s)ide analogues (NAs). However, the dynamics of the HBV‐RNA sequence compared with those of HBV‐DNA during the emergence of antiviral resistance are yet to be elucidated. Methods First, we quantified serum HBV‐RNA in 181 infected patients, and its relationships with clinical characteristics as well as HBV markers were investigated. Next, we undertook simultaneous deep sequencing of HBV‐RNA/HBV‐DNA and their dynamics among four patients receiving NA therapy who were experiencing viral breakthrough. Results Serum HBV‐RNA was detected in 25% (31/123) of cases among patients with HBV without NAs, and the detection rate was significantly high in hepatitis B e antigen‐positive cases with high viral activity. In patients with chronic hepatitis, hepatitis B core‐related antigen was significantly correlated with serum HBV‐RNA irrespective of NA use. In the analysis of the four patients experiencing viral breakthrough, no NA resistance mutation was detected in the serum HBV‐RNA immediately before the breakthrough. However, NA‐resistant sequences appeared at the rates of 0%, 3%, 14%, and 100%, and the NA‐resistant HBV‐RNA sequence rate was correlated with the peak HBV‐DNA titer multiplied by the HBV‐DNA detection duration during the breakthrough (R 2  = 0.978) observed before redisappearance of HBV‐DNA following the addition of new NA. Conclusion Serum HBV‐RNA could reflect the transcriptional activity of covalently closed circular DNA and hepatitis B core‐related antigen. The dynamics of HBV‐RNA could help understanding of the turnover process of HBV covalently closed circular DNA in the liver.
    Type of Medium: Online Resource
    ISSN: 1386-6346 , 1872-034X
    URL: Issue
    URL: Article
    DOI: 10.1111/hepr.v51.1
    DOI: 10.1111/hepr.13574
    Language: English
    Publisher: Wiley
    Publication Date: 2021
    detail.hit.zdb_id: 2006439-1
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  • 4
    Online Resource
    Online Resource
    Spontaneous portosystemic shunt diameter predicts liver function after balloon‐occluded retrograde transvenous obliteration
    Wiley ; 2022
    In:  JGH Open Vol. 6, No. 2 ( 2022-02), p. 139-147
    Details
    In: JGH Open, Wiley, Vol. 6, No. 2 ( 2022-02), p. 139-147
    Abstract: Recently, balloon‐occluded retrograde transvenous obliteration (BRTO), performed for spontaneous portosystemic shunts (SPSS), has been receiving attention as a measure to improve liver function in cirrhotic patients with portal hypertension. However, it is unclear whether SPSS diameter is associated with changes in hepatic venous pressure gradient (HVPG) and liver function after BRTO. Methods In 34 cirrhotic patients receiving BRTO for hepatic encephalopathy/gastric varices, the association of SPSS diameter with liver function at baseline and 6 months after BRTO and the accompanying changes in HVPG were investigated. Results Patients had Child–Pugh (CP) scores of A/B/C (7/19/8), SPSS diameters of ≤10 mm/11–20 mm/ 〈 20 mm (8/21/5), and an average observation period of 3.2 (0.3–8.5) years. SPSS diameter was significantly associated with male sex, alcohol use, and values of albumin, prothrombin time (PT%), and NH 3 at baseline. Moreover, the SPSS diameter was significantly correlated with the changes in HVPG observed upon BRTO ( r  = 0.55, P  = 0.005), and a large shunt diameter was significantly associated with a greater increase in HVPG. At 6 months, significant improvements in albumin, PT%, bilirubin, and NH 3 were observed overall, but the improvement was marked in those with larger shunt diameters if they had CP A/B. Conclusion SPSS diameter was strongly associated with liver function at baseline and after BRTO and also with changes in HVPG, indicating that SPSS diameter is an important predictor of BRTO outcome.
    Type of Medium: Online Resource
    ISSN: 2397-9070 , 2397-9070
    URL: Issue
    URL: Article
    DOI: 10.1002/jgh3.v6.2
    DOI: 10.1002/jgh3.12712
    Language: English
    Publisher: Wiley
    Publication Date: 2022
    detail.hit.zdb_id: 2919809-4
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  • 5
    Online Resource
    Online Resource
    Usefulness of Cell‐Free Human Telomerase Reverse Transcriptase Mutant DNA Quantification in Blood for Predicting Hepatocellular Carcinoma Treatment Efficacy
    Ovid Technologies (Wolters Kluwer Health) ; 2021
    In:  Hepatology Communications Vol. 5, No. 11 ( 2021-11), p. 1927-1938
    Details
    In: Hepatology Communications, Ovid Technologies (Wolters Kluwer Health), Vol. 5, No. 11 ( 2021-11), p. 1927-1938
    Abstract: Although the usefulness of liquid biopsy as a biomarker in the treatment of hepatocellular carcinoma (HCC) has been suggested, its usefulness in transcatheter arterial chemoembolization (TACE) or tyrosine kinase inhibitor (TKI) therapies has not been reported in detail. In this study, we investigated the clinical value of a cell‐free (cf)DNA quantification system targeting the human telomerase reverse transcriptase ( hTERT ) promoter mutation in advanced HCC treatment. Plasma from 67 patients with advanced HCC, treated with TACE and TKI, was used for extraction of cfDNA. We defined cfDNA with the hTERT promoter C228T mutation as circulating mutant DNA (mutant DNA) and without the mutation as circulating wild‐type DNA (wild‐type DNA). We analyzed the changes in mutant and wild‐type DNA levels during HCC treatment and examined the relationship between changes in the cfDNA level and the clinical course. Mutant DNA was detected in 73.1% (49/67) of the patients during HCC treatment. In univariate analysis, factors associated with detection of mutant DNA before treatment were the intrahepatic maximum tumor diameter ( P  = 0.015) and protein induced by vitamin K absence (PIVKAII) ( P  = 0.006). The degree of mutant DNA change after TACE was significantly correlated with tumor volume ( P   〈  0.001), reflecting the treated tumor volume. Responders with peak cfDNA levels within 1 week of TKI initiation had significantly better progression‐free survival than nonresponders ( P  = 0.004). Conclusion: Changes in blood hTERT promoter mutant DNA levels during TACE or TKI treatment indirectly reflect the amount of HCCs and are useful for predicting long‐term treatment responses.
    Type of Medium: Online Resource
    ISSN: 2471-254X , 2471-254X
    URL: Article
    DOI: 10.1002/hep4.1762
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2021
    detail.hit.zdb_id: 2881134-3
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