Conserved properties of hepatitis C genotype 1b proteinsas prognostic markers of response to pegylated interferon and ribavirin combination therapy

Uvod: Infekcija virusom hepatitisa C (HCV) predstavlja značajan globalni zdravstveni problem koji često vodi ka hroničnoj bolesti jetre i cirozi. Prema podacima Svetske zdravstvene organizacije registrovano je preko 170 miliona ljudi inficiranih virusom HCV. Sadašnja standardna terapija hroničnog HCV-a kod pacijenata inficiranih genotipom 1b, koji predstavlja najčešći genotip kako u SAD, zapadnoj Evropi i Srbiji, se sastoji od pegilovanog interferona u kombinaciji sa ribavirinom. Međutim, kombinovana terapija je praćena brojnim neželjenim efektima i dovodi do stabilnog virusološkog odgovora samo kod 50% pacijenata inficiranih genotipom 1. Stoga bi jednostavan i pouzdan test, koji bi pre početka terapije mogao da predvidi virusološki odgovor, bio od velike koristi u kliničkoj praksi. Metode: Identifikacija konzervirane informacije sadržane u proteinima HCV koja korelira sa odgovorom na standardnu kombinovanu terapiju rađena je bioinformatičkom analizom. Uzorci plazme 48 pacijenata sa hroničnom infekcijom HCV, genotipa 1b su klasifikovani u odnosu na odgovor na kombinovanu terapiju. Za određivanje primarne strukture proteina HCV primenjene su klasične metode molekularne biologije: reverzna transkripcija i lančana reakcija polimeraze (Rt-PCR), apsolutna kvantifikacija sa PCR-om u realnom vremenu i automatsko sekvenciranje. Rezultati: Na osnovu rezultata analize svih proteina virusa HCV utvrđeno je da je informacioni sadržaj proteina p7 u korelаciji sа odgovorom nа kombinovаnu terаpiju. Rezultati dobijeni analizom proteinskih sekvenci, 48 pacijenata sa teritorije Srbije su u saglasnosti sa predloženim bioinformatičkim kriterijumom. Posebna pažnja je posvećena optimizaciji eksperimentalnih protokola i formiranju homogenenih grupa u odnosu na osobenosti virusa (tip i podtip) i odgovora na antivirusnu terapiju pacijenata. Zaključak: Na osnovu rezultata ove studije predložen je bioinformatički kriterijum koji omogućаvа procenu odgovora hroničnih HCV bolesnikа inficirаnih genotipom 1b nа kombinovаnu terаpiju.Background: Hepatitis C virus (HCV) infection is a major and rising global health problem, affecting about 170 million people worldwide, according to WHO data, and often leading to chronic liver disease and cirrhosis. The current standard therapy for chronic HCV infection with pegylated interferon combined with ribavirin in patients with the genotype 1 infection, the most frequent genotype in the USA, Western Europe and Serbia, accompanied by numerous side effects, leads to a successful outcome in only about 50% of individuals. Therefore, simple and accurate prediction of hepatitis C treatment response is of great benefit to patients and clinicians. Methods: Identification of the conserved information of the HCV proteins that correlate with the combination therapy outcome was performed by bioinformatics analysis. Plasma samples of 48 chronic HCV patients from Serbia were classified according to the outcome of therapy. To determine primary structure of HCV proteins classical methods of molecular biology: reverse transcription and polymerase chain reaction (Rt-PCR), the absolute quantification-Real Time PCR and DNA sequencing were applied. Results: Among the HCV proteins that we have analyzed the informational property of the p7 of HCV genotype 1b was best related to the therapy outcome. Findings obtained from analyzing sequences 48 patients collected from Serbia were in perfect agreement with proposed bioinformatics criterion. Special attention was paid to optimization experimental protocols and forming homogeneous groups of patients regarding HCV genotype (type and subtype) and therapy response. Conclusions: On the basis of the results in the present study, a simple bioinformatics criterion that could be useful in assessment of the response of HCV-infected patients to the combination therapy has been proposed

Conserved properties of hepatitis C genotype 1b proteinsas prognostic markers of response to pegylated interferon and ribavirin combination therapy

Uvod: Infekcija virusom hepatitisa C (HCV) predstavlja značajan globalni zdravstveni problem koji često vodi ka hroničnoj bolesti jetre i cirozi. Prema podacima Svetske zdravstvene organizacije registrovano je preko 170 miliona ljudi inficiranih virusom HCV. Sadašnja standardna terapija hroničnog HCV-a kod pacijenata inficiranih genotipom 1b, koji predstavlja najčešći genotip kako u SAD, zapadnoj Evropi i Srbiji, se sastoji od pegilovanog interferona u kombinaciji sa ribavirinom. Međutim, kombinovana terapija je praćena brojnim neželjenim efektima i dovodi do stabilnog virusološkog odgovora samo kod 50% pacijenata inficiranih genotipom 1. Stoga bi jednostavan i pouzdan test, koji bi pre početka terapije mogao da predvidi virusološki odgovor, bio od velike koristi u kliničkoj praksi. Metode: Identifikacija konzervirane informacije sadržane u proteinima HCV koja korelira sa odgovorom na standardnu kombinovanu terapiju rađena je bioinformatičkom analizom. Uzorci plazme 48 pacijenata sa hroničnom infekcijom HCV, genotipa 1b su klasifikovani u odnosu na odgovor na kombinovanu terapiju. Za određivanje primarne strukture proteina HCV primenjene su klasične metode molekularne biologije: reverzna transkripcija i lančana reakcija polimeraze (Rt-PCR), apsolutna kvantifikacija sa PCR-om u realnom vremenu i automatsko sekvenciranje. Rezultati: Na osnovu rezultata analize svih proteina virusa HCV utvrđeno je da je informacioni sadržaj proteina p7 u korelаciji sа odgovorom nа kombinovаnu terаpiju. Rezultati dobijeni analizom proteinskih sekvenci, 48 pacijenata sa teritorije Srbije su u saglasnosti sa predloženim bioinformatičkim kriterijumom. Posebna pažnja je posvećena optimizaciji eksperimentalnih protokola i formiranju homogenenih grupa u odnosu na osobenosti virusa (tip i podtip) i odgovora na antivirusnu terapiju pacijenata. Zaključak: Na osnovu rezultata ove studije predložen je bioinformatički kriterijum koji omogućаvа procenu odgovora hroničnih HCV bolesnikа inficirаnih genotipom 1b nа kombinovаnu terаpiju.Background: Hepatitis C virus (HCV) infection is a major and rising global health problem, affecting about 170 million people worldwide, according to WHO data, and often leading to chronic liver disease and cirrhosis. The current standard therapy for chronic HCV infection with pegylated interferon combined with ribavirin in patients with the genotype 1 infection, the most frequent genotype in the USA, Western Europe and Serbia, accompanied by numerous side effects, leads to a successful outcome in only about 50% of individuals. Therefore, simple and accurate prediction of hepatitis C treatment response is of great benefit to patients and clinicians. Methods: Identification of the conserved information of the HCV proteins that correlate with the combination therapy outcome was performed by bioinformatics analysis. Plasma samples of 48 chronic HCV patients from Serbia were classified according to the outcome of therapy. To determine primary structure of HCV proteins classical methods of molecular biology: reverse transcription and polymerase chain reaction (Rt-PCR), the absolute quantification-Real Time PCR and DNA sequencing were applied. Results: Among the HCV proteins that we have analyzed the informational property of the p7 of HCV genotype 1b was best related to the therapy outcome. Findings obtained from analyzing sequences 48 patients collected from Serbia were in perfect agreement with proposed bioinformatics criterion. Special attention was paid to optimization experimental protocols and forming homogeneous groups of patients regarding HCV genotype (type and subtype) and therapy response. Conclusions: On the basis of the results in the present study, a simple bioinformatics criterion that could be useful in assessment of the response of HCV-infected patients to the combination therapy has been proposed

Multiple Roles of the RUNX Gene Family in Hepatocellular Carcinoma and Their Potential Clinical Implications

: Hepatocellular carcinoma (HCC) is one of the most frequent cancers in humans, characterised by a high resistance to conventional chemotherapy, late diagnosis, and a high mortality rate. It is necessary to elucidate the molecular mechanisms involved in hepatocarcinogenesis to improve diagnosis and treatment outcomes. The Runt-related (RUNX) family of transcription factors (RUNX1, RUNX2, and RUNX3) participates in cardinal biological processes and plays paramount roles in the pathogenesis of numerous human malignancies. Their role is often controversial as they can act as oncogenes or tumour suppressors and depends on cellular context. Evidence shows that deregulated RUNX genes may be involved in hepatocarcinogenesis from the earliest to the latest stages. In this review, we summarise the topical evidence on the roles of RUNX gene family members in HCC. We discuss their possible application as non-invasive molecular markers for early diagnosis, prognosis, and development of novel treatment strategies in HCC patients

Genetic Polymorphisms of Neurocardiovascular Disorders

The autonomic nervous control of cardiovascular (CV) system plays a major role in the adaptation of the organism to the changes in external and internal environment. It’s dysfunction is the major pathophysiological factor in the development of neurocardiovascular diseases. The aim of this review is to present the state of the art on the role of candidate gene polymorphisms of the molecules in the signaling chain of neurocardiovascular transmission in neurocardiovascular diseases. Neurocardiovascular disorders can be classified as sympathetic vs. vagally mediated disorders, though in many disorders both systems are dysfunctional. A number of molecules along the signaling pathway can be functionally modified and be the background of the predisposition, faster progression or complicated form of the disease. When the disease is the consequence of the joined parasympathetic and sympathetic nervous system disequilibrium, the focus of neurogenetic research should be on molecules providing the cross-talk between the two systems: on intercellular and intracellular level and on the level of the signaling process integration. An aggregation of positive results for the association between certain genes and different neurocardiovascular phenotypes pointed on a specific "neurocardiovascular genetic hotspots". Identification of these genes could be of particular interest as a diagnostic tool in the clustered form of neurocardiovascular diseases. New data obtained from neurogenetic approach will improve the disease outcome by gene, pharmacologic and behavioral modulation of the autonomic nervous system

Hepatitis C virus as cause of fulminant hepatitis-sequence analysis of the 5’ nontranslated region

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ABO histo-blood groups and Rh systems in relation to malignant tumors of the digestive tract in Bosnia and Herzegovina

The distribution of ABO blood groups and the Rhesus factor was analyzed in 279 patients who suffered from malignant tumors of the digestive system. Patients were registered retrospectively in the Gastroenterohepatology Clinic, Clinical Center, University of Sarajevo over a discontinuous period of 88 months. From the results obtained, it was concluded that: (a) men became ill from gastric cancer significantly more frequently than women; (b) the frequency of liver carcinoma was three times higher than the global frequency and the frequency neighboring ethnic groups; and (c) patients with blood group B and patients with RhD(-) exhibited a significantly higher proportion of disease.Analizirana je distribucija krvnih grupa ABO sistema i Rezus faktora kod 279 pacijenata obolelih od malignih tumora digestivnog sistema. Pacijenti su registrovani retrospektivno u Gastroenterohepatološkoj klinici Kliničkog centra Univerziteta u Sarajevu u diskontinuitetu tokom 88 meseci (1987-1998). Na osnovu analizirane populacije pacijenata zaključeno je da: (a) muškarci značajno češće oboljevaju od kancera želuca u odnosu na žene; (b) učestalost karcinoma jetre je tri puta veća upoređenju sa učestalošću ovog oboljenja u svetu i susednim zemljama; (c) pacijenti sa V krvnom grupom i pacijenti sa RhD(-) su u značajnom stepenu češći u ispitivanoj populaciji obolelih od očekivanog.Projekat ministarstva br. 143010 i 14301

Variability of the HCV core region and host genetic and epigenetic factors can predict the response to pegylated interferon/ribavirin therapy in genotype 1b hepatitis C patients from Serbia

Variations in the hepatitis C virus (HCV) core sequence have been related to disease progression and response to antiviral therapy. Previously we showed that the methylation status of RASSF1A and p16 genes, and IL28B genotypes affects the response to pegylated interferon/ribavirin (PEG-IFN/RBV) therapy. Herein we investigated whether amino acid (aa) substitutions in the HCV core region alone or in combination with IL28B genotypes and RASSF1A/p16 methylation affect the response to PEG-IFN/RBV therapy and liver disease progression. Among 29 examined patients, we found no association between single aa substitutions and response to therapy. However, we observed that patients with the HCV core aa substitution at position 75 and CT/TT IL28B genotypes were non-responders (NR), (P=0.023). Moreover, these patients had unmethylated RASSF1A. In contrast, most patients (75%) with aa substitutions at position 91 and CC IL28B genotype achieved sustained virologic response (SVR), (P=0.030), and 70% of them had methylated RASSF1A gene. Our results suggest that combined analysis of aa substitutions in the core protein, the IL28B rs12979860 polymorphism, and the methylation status of the RASSF1A gene may help in predicting treatment response to PEG-IFN/RBV in genotype 1b chronic hepatitis C patients

Sequence variability at the internal ribosome entry site of the HCV genome in relation to therapy outcome

Different types of interferon are widely used to treat hepatitis C virus (HCV) infection. Results obtained in vitro suggest that interferon inhibits internal ribosome entry site (IRES)-mediated translation of the HCV genome. To elucidate the possible effect of the nucleotide sequence of IRES on therapy outcome, we compared HCV isolates from patients with sustained response and non-response to interferon/ribavirin combination therapy. In 56 analyzed HCV isolates, nucleotide changes appeared strictly in the stem-loop IIIb region, the stem part from 243 nt to 248 nt, and the polypyrimidine-II region. The natural sequence variability of IRES in isolates of genotype 3a was significantly higher than in isolates of genotype 1b (p < 0.05). The average number of nucleotide changes in genotype 3a correlated with response to therapy (p < 0.05).Interferonska terapija se danas najčešće koristi u lečenju infekcije virusom hepatitisa tipa C (HCV). In vitro rezultati su pokazali da interferoninhibira translaciju kod ovog virusa preko interakcije sa delom genoma koji učestvuje u inicijaciji translacije tzv. 'unutrašnje ribozomalno ulazno mesto' (IRES). U ovom radu smo ispitivali nukleotidne izmene u IRES-u kod izolata HCV-a dobijenih iz seruma osoba koje su primale kombinovanu terapiju interferon/ribavirin. U analiziranoj grupi od 56 HCV izolata, nukleotidne izmene su utvrđene u: IIIb petlji, regiji između 243 nt i 248 nt i polipirimidin-II regiji. Utvrđena varijabilnost IRES-a kod izolata genotipa 3a značajno je veća u poređenju sa izolatima genotipa 1b (p < 0.05). Prosečan broj nukleotidnih izmena kod izolata genotipa 3a je u korelaciji sa odgovorom na primenjenu terapiju (p < 0.05).Projekat ministarstva br. 143010 i 14301

ABO histo-blood groups and Rh systems in relation to malignant tumors of the digestive tract in Bosnia and Herzegovina

The distribution of ABO blood groups and the Rhesus factor was analyzed in 279 patients who suffered from malignant tumors of the digestive system. Patients were registered retrospectively in the Gastroenterohepatology Clinic, Clinical Center, University of Sarajevo over a discontinuous period of 88 months. From the results obtained, it was concluded that: (a) men became ill from gastric cancer significantly more frequently than women; (b) the frequency of liver carcinoma was three times higher than the global frequency and the frequency neighboring ethnic groups; and (c) patients with blood group B and patients with RhD(-) exhibited a significantly higher proportion of disease.Analizirana je distribucija krvnih grupa ABO sistema i Rezus faktora kod 279 pacijenata obolelih od malignih tumora digestivnog sistema. Pacijenti su registrovani retrospektivno u Gastroenterohepatološkoj klinici Kliničkog centra Univerziteta u Sarajevu u diskontinuitetu tokom 88 meseci (1987-1998). Na osnovu analizirane populacije pacijenata zaključeno je da: (a) muškarci značajno češće oboljevaju od kancera želuca u odnosu na žene; (b) učestalost karcinoma jetre je tri puta veća upoređenju sa učestalošću ovog oboljenja u svetu i susednim zemljama; (c) pacijenti sa V krvnom grupom i pacijenti sa RhD(-) su u značajnom stepenu češći u ispitivanoj populaciji obolelih od očekivanog.Projekat ministarstva br. 143010 i 14301