Microarrays-Enabled Hypothesis Generation: The Suspect Role of FNBP-1 in Neuropsychiatric Pathogenesis Associated with HIV and/or HCV Infection.

OBJECTIVE: The spectrum of neuropsychiatric illness (NI) associated with the Human Immunodeficiency Virus (HIV) and/or the Hepatitis C Virus (HCV) is far reaching and significantly impacts the clinical presentation and outcome of infected persons; however, the etiological and pathophysiological background remains partially understood. The present work was aimed to investigate the potential significance of formin binding protein 1 (FNBP-1)-dependent pathways in NI-pathogenesis by elaborating on previous microarray-based research in HIV and/or HCV-infected patients receiving interferon-α (IFN-α) immunotherapy via a rigorous data mining procedure. METHODS: Using microarray data of peripheral whole blood (PB) samples obtained from HCV mono-infected persons (n=25, Affymetrix® HG-U133A_2) 12 h before and after the 1st dose of pegylated IFN-α (PegIFN-α), we re-applied the same analytical algorithm that we had developed and published in an earlier study with HIV/HCV co-infected subjects (N=28, Affymetrix® HG-U133A), in order to evaluate reproducibility of potential NI-related molecular findings in an independent cohort. RESULTS: Among 28 gene expression profiles (HIV/HCV: N=9 vs. HCV: N=19) selected by applying different thresholds (a Mean Fold Difference value (MFD) in gene expression of ≥ 0.38 (log2) and/or P value from <0.05 to ≤ 0.1) FNBP-1 was identified as the only overlapping marker, which also exhibited a consistent upregulation in association with the development of NI in both cohorts. Previous functional annotation analysis had classified FNBP-1 as molecule with significant enrichment in various brain tissues (P<0.01). CONCLUSION: Our current findings are strongly arguing for intensifying research into the FNBP-1-related mechanisms that may be conferring risk for or resistance to HIV- and/or HCV-related NI.

CCL5 mRNA is a marker for early fibrosis in chronic hepatitis C and is regulated by interferon-α therapy and toll-like receptor 3 signalling.

Mechanisms causing liver fibrosis during chronic hepatitis C virus infection (cHCV) are not sufficiently understood. This study was aimed to identify biomarkers for early fibrosis (EF) and to investigate their potential role in cHCV-related fibrogenesis. To this end, peripheral whole blood (PB) samples from 36 patients with cHCV recruited from two independent cohorts were subjected to microarray analysis 12 h before initiation of peginterferon-alpha (Peg-IFN-α) and ribavirin therapy. Liver biopsies were evaluated using the Batts-Ludwig staging (BL-S) classification system for fibrosis. We showed that gene expression profiles (N = 8) distinguished between EF (BL-S: 0,1) and late fibrosis (LF; BL-S: 2,3,4) with 88.9% accuracy. Fibrosis-related functional annotations for chemokine-'C-C-motif'' ligand 5 (CCL5) provided foundation for focused investigation, and qRT-PCR confirmed that CCL5 mRNA levels (PB) reliably discriminate EF from LF (accuracy: 86.7%). Positive correlations (P < 0.05) with CCL5 mRNA levels and EF discovered gene expression profiles (PB) reflecting stable expression of IFN-α receptor 1, negative regulation of the MyD88-dependent toll-like receptor (TLR) pathway and decreased expression of TLR3 in vivo. Remarkably, Peg-IFN-α suppressed CCL5 mRNA levels (PB) in EF in vivo. These findings along with results from parallel in vitro investigation into the effect of IFN-α or poly I:C (TLR3-agonist) on CCL5 gene expression in hepatic stellate cells (HSC) attest to the multi-site involvement of these pathways in regulating fibrogenesis. In conclusion, we identified novel, reliable biomarkers for EF and exposed functional properties of the molecular network regulating CCL5 biosynthesis in peripheral or hepatic cell types with key roles in cHCV-related liver and/or immune pathogenesis.

Core promoter mutant HBV non-responding to adefovir after viral breakthrough on lamivudine: rapid virologic response to tenofovir plus lamivudine in a cirrhotic patient.

BACKGROUND: In chronic hepatitis B patients undergoing therapy with LAM or ADV, viral breakthrough is possible due to the emergence of drug resistance. LAM resistant HBV strains are susceptible to ADV, while ADV resistant mutants remain sensitive to LAM. CASE REPORT: A male patient with HBV-related cirrhosis developed viral breakthrough (HBV DNA>1.8 x 106 IU/ml) after 4 1/2 years of treatment with LAM, and therapy was switched to ADV (10 mg/d). After three months, HBV remained highly replicative without any changes of ALT values, and ADV dose was increased (20 mg/d). Because of unchanged VL sequence analysis was performed three months later, which showed the mutation (rtS219A) and the concomitant mutation (sS210R) and 2 mutations in core promoter region (A1762T), (G1764A). During the sixth month of ADV monotherapy the patient developed liver failure. After administration of TDF plus LAM, HBV DNA became undetectable within 39 days. At day 41, the patient underwent OLT. TDF plus LAM were well tolerated, and the patient maintained undetectable HBV DNA levels, and in addition to HBIG a sustained HBsAg negative status over twenty-eight months post OLT. CONCLUSION: TDF plus LAM is a safe drug combination in case of viral breakthrough during LAM treatment and subsequent primary non-response to ADV. High VL persisting for >or= 6 months of continuous antiviral treatment may indicate drug resistance. Especially in cirrhotic patients with LAM resistance, "add on" of a nucleotide analogue is the right therapeutic strategy even before viral breakthrough gets apparent.

Efavirenz-therapy in HIV-patients with underlying liver disease: importance of continuous TDM of EFV.

OBJECTIVE: Many HIV-patients have a chronic liver disease due to HBV-/HCV-coinfections and/or consume of alcohol. In these patients therapy with EFV is often problematic because of NNRTI associated liver toxicity. Measurement of EFV plasmalevels and dose adjustment using TDM should be evaluated in this study. METHODS: EFV-plasmasamples were standardized drawn 12h after ingestion. Measurement of 576 EFV plasmalevels was performed by HPLC. EFV plasmalevels as well as ALT-, AST- and GGT-values of 64 patients treated with EFV (5206 weeks) were measured regularly. 16 patients had a HCV-coinfection, 3 had a HBV-coinfection and 5 had an concomitant alcoholic liver disease. Maximal changes of ALT-, AST- and GGT-values (DeltaALT, DeltaAST, DeltaGGT), CD4-/CD8 cells and HIV-RNA were registered during therapy. Dose adjustment was performed for EFV plasmalevels out of target range 1000-4000 ng/ml. RESULTS: EFV plasmalevels of 40 HIV-patients (2288 +/- 1199 ng/ml) showed no significant differences compared to plasmalevels of HIV/HCV-patients (2391 +/- 976 ng/ml) or to plasmalevels of HIV/HBV-patients (1913 +/- 288 ng/ml) or to those of HIV-patients with alcoholic liver disease (1702 +/- 506 ng/ml). In 24 HIV-patients with underlying liver disease median DeltaGGT was +25 IU/l, median DALT was +13 IU/l and median DeltaAST was +8 IU/l. Dose adjustment was performed in 1 patient during study period. Increasing rates of ALT-, AST- and GGT-values showed no significant differences between liver healthy HIV-patients and those with a liver disease. 44 patients with continuous EFV plasmalevels in target range reached a viral suppression <100 c/ml during therapy. CONCLUSIONS: EFV-plasmalevels of HIV-infected patients showed no significant differences compared to EFV-plasmalevels of coinfected patients with concomitant liver disease. In those patients DeltaALT, DeltaAST and DeltaGGT were not significantly different than in liver-healthy HIV-patients with normal EFV-plasmaconcentrations. EFV plasmalevels in target range of 1000-4000 ng/ml correlate to a good viral response. One patient after dose adjustment was able to continue therapy. Using TDM EFV therapy in patients with underlying liver disease is save.