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  • Ovid Technologies (Wolters Kluwer Health)  (2)
  • Maekawa, Shinya  (2)
  • 1
    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
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2022
    detail.hit.zdb_id: 2881134-3
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  • 2
    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
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2021
    detail.hit.zdb_id: 2881134-3
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
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