Hepatitis B-associated hepatocellular carcinoma (HBV-HCC) remains an intractable high-mortality solidtumor cancer that accounted for 42% of global HCC cases in 2019. Despite some developments in systemic therapy,only ...Hepatitis B-associated hepatocellular carcinoma (HBV-HCC) remains an intractable high-mortality solidtumor cancer that accounted for 42% of global HCC cases in 2019. Despite some developments in systemic therapy,only a small subset of late-stage HCC patients responds positively to recently developed therapeutic innovations.MicroRNAs (miRNAs) act as an ancillary epigenetic system that can regulate genome expression in all cancerpathways including HCC. The molecular mechanisms of miRNA regulation in cancer pathogenesis offered researchersa new approach that was widely hoped would translate into miRNA-based drugs and diagnostics. Thirty years on,miRNA-based diagnostic and therapeutic agents for HCC remain a work-in-progress (WIP) and no current miRNAHCC clinical trial has progressed to Phase 4. The question remains why this is the case after 30 years and what is theway forward. The major findings and contribution of this paper are that it illustrates the complexity of the HBVmiRNA interactome in HBV-HCC in all cellular processes, as well as the ancillary role of miRNA in the epigeneticand immune systems. This is combined with a review of the outcomes and problems of clinical trials, to explain whymiRNA therapeutics and diagnostics have not progressed to approved drugs or serum-based diagnostic tests. The wayforward suggests a radical rethink might be so that involves the incorporation of AI, bioinformatics, andnanotechnology to solve the problem.展开更多
AIM： To examine the serum from black African patients with acute hepatitis B to ascertain if integrants of viral DNA can be detected in fragments of cellular DNA leaking from damaged hepatocytes into the circulation....AIM： To examine the serum from black African patients with acute hepatitis B to ascertain if integrants of viral DNA can be detected in fragments of cellular DNA leaking from damaged hepatocytes into the circulation. METHODS： DNA was extracted from the sera of five patients with uncomplicated acute hepatitis B and one with fulminant disease. Two subgenomic PCRs designed to amplify the complete genome of HBV were used and the resulting amplicons were cloned and sequenced. RESULTS： HBV and chromosomal DNA were amplified from the sera of all the patients. In one patient with uncomplicated disease, HBV DNA was integrated into host chromosome 7 q11.23 in the WBSCR1 gene. The viral DNA comprised 200 nucleotides covering the S and X genes in opposite orientation, with a 1 169 nucleotide deletion. The right virus/host junction was situated at nucleotide 1 774 in the cohesive overlap region of the viral genome, at a preferred topoisomerase I cleavage motif. The chromosomal DNA was not rearranged. The patient made a full recovery and seroconverted to anti-HBs- and anti-HBe-positivity. Neither HBV nor chromosomal DNA could be amplified from his serum at that time. CONCLUSION： Integration of viral DNA into chromosomal DNA may occur rarely during acute hepatitis B and, with clonal propagation of the integrant, might play a role in hepatocarcinogenesis.展开更多
AIM: To determine whether one genotype (A or non-A genotypes of HBV) predominated over the other during the course of HBV infection.METHODS: Four baboons were inoculated with HBV. DNA was extracted from serum obtained...AIM: To determine whether one genotype (A or non-A genotypes of HBV) predominated over the other during the course of HBV infection.METHODS: Four baboons were inoculated with HBV. DNA was extracted from serum obtained at monthly intervals postinoculation for 52 weeks and HBV DNA was amplified using primers specific for the core region containing an insert characteristic of genotype A (nt 2 354-2 359, numbering from the EcoRI site). The amplicons were cloned into PCRScriptTM and a minimum of 15 clones per time point were sequenced in both directions.RESULTS: Both genotypes persisted for the entire followup period of 52 weeks. Genotype non-A predominated in two baboons and genotype A in one baboon. Neither genotype predominated in the fourth baboon, as shown at a 5 % level of testing.CONCLUSION: No conclusions concerning the dominance of either genotype or the natural progression or replication rates of HBV could be drawn because the pattern of the genotypes found may have been caused by sampling fluctuations at the time of DNA extraction and cloning as a result of the very low viral loads in the baboon sera.展开更多
Hepatitis B virus(HBV)(sub)genotypes A1,D3 and E circulate in sub-Saharan Africa,the region with one of the highest incidences of HBV-associated hepatocellular carcinoma globally.Although genotype E was identified mor...Hepatitis B virus(HBV)(sub)genotypes A1,D3 and E circulate in sub-Saharan Africa,the region with one of the highest incidences of HBV-associated hepatocellular carcinoma globally.Although genotype E was identified more than 20 years ago,and is the most widespread genotype in Africa,it has not been extensively studied.The current knowledge status and gaps in its origin and evolution,natural history of infection,disease progression,response to antiviral therapy and vaccination are discussed.Genotype E is an African genotype,with unique molecular characteristics that is found mainly in Western and Central Africa and rarely outside Africa except in individuals of African descent.The low prevalence of this genotype in the African descendant populations in the New World,phylogeographic analyses,the low genetic diversity and evidence of remnants of genotype E in ancient HBV samples suggests the relatively recent reintroduction into the population.There is scarcity of information on the clinical and virological characteristics of genotype E-infected patients,disease progression and outcomes and efficacy of anti-HBV drugs.Individuals infected with genotype E have been characterised with high hepatitis B e antigen-positivity and high viral load with a lower end of treatment response to interferon-alpha.A minority of genotype E-infected participants have been included in studies in which treatment response was monitored.Of concern is that current guidelines do not consider patients infected with genotype E.Thus,there is an urgent need for further large-scale investigations into genotype E,the neglected genotype of HBV.展开更多
AIM:To molecularly characterize hepatitis B virus(HBV)isolates from Kerala and to relate them to the clinical manifestation of infection.METHODS:Sera and clinical data were collected from91 patients diagnosed with chr...AIM:To molecularly characterize hepatitis B virus(HBV)isolates from Kerala and to relate them to the clinical manifestation of infection.METHODS:Sera and clinical data were collected from91 patients diagnosed with chronic HBV infection and HBV-related hepatocellular carcinoma(HCC).HBV from44 HCC,22 cirrhotic and 25 chronic hepatitis patients were genotyped by sequencing of the complete S region or by restriction fragment length polymorphism assays.The basic core promoter/precore region was sequenced.The complete surface DNA sequences were assembled and aligned manually,and then compared with the sequences of HBV of genotypes(A-J)from GenBank.The evolutionary history was inferred using the Neighbor-Joining method and the evolutionary distances computed using the Kimura 2-parameter method.Bootstrapping was performed using 1000 replicates.The TaqMan BS-1 probe was used to quantify HBV DNA at a lower detection limit of approximately20 IU/mL.Continuous variables were compared using an independent Student’s t test.Theχ2test or Fisher’s exact test was used to compare categorical variables.The differences were considered statistically significant at P<0.05.RESULTS:Irrespective of disease status,the predominant genotype was A(72%);95%belonging to subgenotype A1,followed by genotypes D(27%)and C(1%).HCC patients infected with subgenotype A1were significantly younger than those infected with D.Mutation A1762T/G1764A was significantly associated with HCC in both genotypes A and D.Mutation G1862T was more frequent in subgenotype A1(P<0.0001),and in combination with A1762T/G1764A,it was significantly associated with HBV from HCC patients.Mutation C1766T/T1768A was significantly associated with genotype A(P=0.05)and HCC(P=0.03).The preS2start codon M1T/I mutation was unique to genotype A strains(15.6%)from all disease groups and occurred at a higher frequency in isolates from HCC patients(P=0.076).A higher frequency of preS deletion mutants(33.3%)was observed in genotype A from HCC compared with non-HCC patients,but did not reach statistical significance.The preS2:F22L mutation was found in genotypes A and D.CONCLUSION:Kerala is the first Indian state in which subgenotype A1 has been found to predominate in liver disease patients who developed HCC at a relatively young age.展开更多
With the highest annual fatality ratio(mortality-to-incidence ratio),reported for a human cancer,hepatocellular carcinoma(HCC)ranks as the third leading cause of cancer-related deaths worldwide and its distribution is...With the highest annual fatality ratio(mortality-to-incidence ratio),reported for a human cancer,hepatocellular carcinoma(HCC)ranks as the third leading cause of cancer-related deaths worldwide and its distribution is not uniform.In Sub-Saharan Africa(SSA),HCC is the second leading cause of cancer-related deaths for men and the fourth for women in 2020,with average age-standardised mortality rates of 8.2 and 4.2 per 100,000 persons/year,respectively.In this region,HCC presents in younger age groups and has a median survival rate of~3-4 months.The major risk factors for HCC include viral[hepatitis B virus(HBV),hepatitis C virus(HCV)and hepatitis D virus(HDV)]and environmental[dietary aflatoxin and iron overload]factors,with more than 50%being attributable to HBV,which is endemic in SSA.HCC control efforts in SSA are faced with a number of unique challenges,including resource restrictions,a paucity of good data,few cancer registries,inaccessibility of treatment for HBV and HCV,co-infection with human immunodeficiency virus(HIV),exposure to co-carcinogen aflatoxin B1,unique(sub)genotypes of HBV and changing natural history and aetiology of HCC as a result of antiretroviral therapy rollout for HIV and changing lifestyles.These unique features of HCC in SSA,together with the challenges faced in its prevention and appropriate public health intervention,diagnosis and treatment,all suggest that HCC in SSA is deserving of an in depth understanding by further focused research.Considerable motivation of policymakers,work and resources are required to reduce the burden of this cancer on the subcontinent.展开更多
The increasing incidence and mortality of hepatocellular carcinoma(HCC)driven by hepatitis B virus(HBV)infection is a global public health concern[1].Despite World Health Organization(WHO)targets for the elimination o...The increasing incidence and mortality of hepatocellular carcinoma(HCC)driven by hepatitis B virus(HBV)infection is a global public health concern[1].Despite World Health Organization(WHO)targets for the elimination of hepatitis as a public health problem by 2030,death attributable to viral hepatitis has continued to rise over the past decade[2],in contrast to the reduction seen in the comparable threats of human immunodeficiency virus(HIV),tuberculosis(TB)and malaria.展开更多
文摘Hepatitis B-associated hepatocellular carcinoma (HBV-HCC) remains an intractable high-mortality solidtumor cancer that accounted for 42% of global HCC cases in 2019. Despite some developments in systemic therapy,only a small subset of late-stage HCC patients responds positively to recently developed therapeutic innovations.MicroRNAs (miRNAs) act as an ancillary epigenetic system that can regulate genome expression in all cancerpathways including HCC. The molecular mechanisms of miRNA regulation in cancer pathogenesis offered researchersa new approach that was widely hoped would translate into miRNA-based drugs and diagnostics. Thirty years on,miRNA-based diagnostic and therapeutic agents for HCC remain a work-in-progress (WIP) and no current miRNAHCC clinical trial has progressed to Phase 4. The question remains why this is the case after 30 years and what is theway forward. The major findings and contribution of this paper are that it illustrates the complexity of the HBVmiRNA interactome in HBV-HCC in all cellular processes, as well as the ancillary role of miRNA in the epigeneticand immune systems. This is combined with a review of the outcomes and problems of clinical trials, to explain whymiRNA therapeutics and diagnostics have not progressed to approved drugs or serum-based diagnostic tests. The wayforward suggests a radical rethink might be so that involves the incorporation of AI, bioinformatics, andnanotechnology to solve the problem.
基金Supported by grants from the Poliomyelitis Research Foundation of South African and the HE Griffin Cancer Trust
文摘AIM： To examine the serum from black African patients with acute hepatitis B to ascertain if integrants of viral DNA can be detected in fragments of cellular DNA leaking from damaged hepatocytes into the circulation. METHODS： DNA was extracted from the sera of five patients with uncomplicated acute hepatitis B and one with fulminant disease. Two subgenomic PCRs designed to amplify the complete genome of HBV were used and the resulting amplicons were cloned and sequenced. RESULTS： HBV and chromosomal DNA were amplified from the sera of all the patients. In one patient with uncomplicated disease, HBV DNA was integrated into host chromosome 7 q11.23 in the WBSCR1 gene. The viral DNA comprised 200 nucleotides covering the S and X genes in opposite orientation, with a 1 169 nucleotide deletion. The right virus/host junction was situated at nucleotide 1 774 in the cohesive overlap region of the viral genome, at a preferred topoisomerase I cleavage motif. The chromosomal DNA was not rearranged. The patient made a full recovery and seroconverted to anti-HBs- and anti-HBe-positivity. Neither HBV nor chromosomal DNA could be amplified from his serum at that time. CONCLUSION： Integration of viral DNA into chromosomal DNA may occur rarely during acute hepatitis B and, with clonal propagation of the integrant, might play a role in hepatocarcinogenesis.
文摘AIM: To determine whether one genotype (A or non-A genotypes of HBV) predominated over the other during the course of HBV infection.METHODS: Four baboons were inoculated with HBV. DNA was extracted from serum obtained at monthly intervals postinoculation for 52 weeks and HBV DNA was amplified using primers specific for the core region containing an insert characteristic of genotype A (nt 2 354-2 359, numbering from the EcoRI site). The amplicons were cloned into PCRScriptTM and a minimum of 15 clones per time point were sequenced in both directions.RESULTS: Both genotypes persisted for the entire followup period of 52 weeks. Genotype non-A predominated in two baboons and genotype A in one baboon. Neither genotype predominated in the fourth baboon, as shown at a 5 % level of testing.CONCLUSION: No conclusions concerning the dominance of either genotype or the natural progression or replication rates of HBV could be drawn because the pattern of the genotypes found may have been caused by sampling fluctuations at the time of DNA extraction and cloning as a result of the very low viral loads in the baboon sera.
基金Supported by National Research Foundation(NRF)of South Africa,No.GUN#93516Deutsche Forschungsgemeinschaft(DFG,English:German Research Foundation)Cancer Association of South Africa(CANSA).
文摘Hepatitis B virus(HBV)(sub)genotypes A1,D3 and E circulate in sub-Saharan Africa,the region with one of the highest incidences of HBV-associated hepatocellular carcinoma globally.Although genotype E was identified more than 20 years ago,and is the most widespread genotype in Africa,it has not been extensively studied.The current knowledge status and gaps in its origin and evolution,natural history of infection,disease progression,response to antiviral therapy and vaccination are discussed.Genotype E is an African genotype,with unique molecular characteristics that is found mainly in Western and Central Africa and rarely outside Africa except in individuals of African descent.The low prevalence of this genotype in the African descendant populations in the New World,phylogeographic analyses,the low genetic diversity and evidence of remnants of genotype E in ancient HBV samples suggests the relatively recent reintroduction into the population.There is scarcity of information on the clinical and virological characteristics of genotype E-infected patients,disease progression and outcomes and efficacy of anti-HBV drugs.Individuals infected with genotype E have been characterised with high hepatitis B e antigen-positivity and high viral load with a lower end of treatment response to interferon-alpha.A minority of genotype E-infected participants have been included in studies in which treatment response was monitored.Of concern is that current guidelines do not consider patients infected with genotype E.Thus,there is an urgent need for further large-scale investigations into genotype E,the neglected genotype of HBV.
基金Supported by The National Research Foundation of South Africa,NRF,GUN 65530(to Kramvis A)and the Cancer Associationof South Africapostdoctoral funding from the NRF,GUN 75055 and the University of the Witwatersrand(to Gopalakrishnan D)bursaries from the University of the Witwatersrand,the Poliomyelitis Research Foundation and the Ernst and Ethel Eriksen Trust(to Keyter M)
文摘AIM:To molecularly characterize hepatitis B virus(HBV)isolates from Kerala and to relate them to the clinical manifestation of infection.METHODS:Sera and clinical data were collected from91 patients diagnosed with chronic HBV infection and HBV-related hepatocellular carcinoma(HCC).HBV from44 HCC,22 cirrhotic and 25 chronic hepatitis patients were genotyped by sequencing of the complete S region or by restriction fragment length polymorphism assays.The basic core promoter/precore region was sequenced.The complete surface DNA sequences were assembled and aligned manually,and then compared with the sequences of HBV of genotypes(A-J)from GenBank.The evolutionary history was inferred using the Neighbor-Joining method and the evolutionary distances computed using the Kimura 2-parameter method.Bootstrapping was performed using 1000 replicates.The TaqMan BS-1 probe was used to quantify HBV DNA at a lower detection limit of approximately20 IU/mL.Continuous variables were compared using an independent Student’s t test.Theχ2test or Fisher’s exact test was used to compare categorical variables.The differences were considered statistically significant at P<0.05.RESULTS:Irrespective of disease status,the predominant genotype was A(72%);95%belonging to subgenotype A1,followed by genotypes D(27%)and C(1%).HCC patients infected with subgenotype A1were significantly younger than those infected with D.Mutation A1762T/G1764A was significantly associated with HCC in both genotypes A and D.Mutation G1862T was more frequent in subgenotype A1(P<0.0001),and in combination with A1762T/G1764A,it was significantly associated with HBV from HCC patients.Mutation C1766T/T1768A was significantly associated with genotype A(P=0.05)and HCC(P=0.03).The preS2start codon M1T/I mutation was unique to genotype A strains(15.6%)from all disease groups and occurred at a higher frequency in isolates from HCC patients(P=0.076).A higher frequency of preS deletion mutants(33.3%)was observed in genotype A from HCC compared with non-HCC patients,but did not reach statistical significance.The preS2:F22L mutation was found in genotypes A and D.CONCLUSION:Kerala is the first Indian state in which subgenotype A1 has been found to predominate in liver disease patients who developed HCC at a relatively young age.
文摘With the highest annual fatality ratio(mortality-to-incidence ratio),reported for a human cancer,hepatocellular carcinoma(HCC)ranks as the third leading cause of cancer-related deaths worldwide and its distribution is not uniform.In Sub-Saharan Africa(SSA),HCC is the second leading cause of cancer-related deaths for men and the fourth for women in 2020,with average age-standardised mortality rates of 8.2 and 4.2 per 100,000 persons/year,respectively.In this region,HCC presents in younger age groups and has a median survival rate of~3-4 months.The major risk factors for HCC include viral[hepatitis B virus(HBV),hepatitis C virus(HCV)and hepatitis D virus(HDV)]and environmental[dietary aflatoxin and iron overload]factors,with more than 50%being attributable to HBV,which is endemic in SSA.HCC control efforts in SSA are faced with a number of unique challenges,including resource restrictions,a paucity of good data,few cancer registries,inaccessibility of treatment for HBV and HCV,co-infection with human immunodeficiency virus(HIV),exposure to co-carcinogen aflatoxin B1,unique(sub)genotypes of HBV and changing natural history and aetiology of HCC as a result of antiretroviral therapy rollout for HIV and changing lifestyles.These unique features of HCC in SSA,together with the challenges faced in its prevention and appropriate public health intervention,diagnosis and treatment,all suggest that HCC in SSA is deserving of an in depth understanding by further focused research.Considerable motivation of policymakers,work and resources are required to reduce the burden of this cancer on the subcontinent.
基金funding from the Wellcome Trust(Grant ref 110110/Z/15/Z)the Francis Crick Institute,London.
文摘The increasing incidence and mortality of hepatocellular carcinoma(HCC)driven by hepatitis B virus(HBV)infection is a global public health concern[1].Despite World Health Organization(WHO)targets for the elimination of hepatitis as a public health problem by 2030,death attributable to viral hepatitis has continued to rise over the past decade[2],in contrast to the reduction seen in the comparable threats of human immunodeficiency virus(HIV),tuberculosis(TB)and malaria.
