Expression of Biliverdin Reductase A in peripheral blood leukocytes is associated with treatment response in HCV-infected patients

Iva Subhanova, Lucie Muchova, Martin Lenicek, Hendrik J Vreman, Ondrej Luksan, Kristyna Kubickova, Miluse Kreidlova, Tomas Zima, Libor Vitek, Petr Urbanek, Iva Subhanova, Lucie Muchova, Martin Lenicek, Hendrik J Vreman, Ondrej Luksan, Kristyna Kubickova, Miluse Kreidlova, Tomas Zima, Libor Vitek, Petr Urbanek

Abstract

Background and aims: Hepatitis C virus (HCV) infection is associated with systemic oxidative stress. Since the heme catabolic pathway plays an important role in antioxidant protection, we attempted to assess the gene expression of key enzymes of heme catabolism, heme oxygenase 1 (HMOX1), heme oxygenase 2 (HMOX2), and biliverdin reductase A (BLVRA) in the liver and peripheral blood leukocytes (PBL) of patients chronically infected with HCV.

Methods: Gene expressions (HMOX1, HMOX2, BLVRA) and HCV RNA were analyzed in PBL of HCV treatment naïve patients (n = 58) and controls (n = 55), with a subset of HCV patients having data on hepatic gene expression (n = 35). Based upon the therapeutic outcome, HCV patients were classified as either responders (n = 38) or treatment-failure patients (n = 20). Blood samples in HCV patients were collected at day 0, and week 12, 24, 36, and 48 after the initiation of standard antiviral therapy.

Results: Compared to the controls, substantially increased BLVRA expression was detected in PBL (p<0.001) of therapeutically naïve HCV patients. mRNA levels of BLVRA in PBL closely correlated with those in liver tissue (r2 = 0.347,p = 0.03). A marked difference in BLVRA expression in PBL between the sustained responders and patients with treatment failure was detected at week 0 and during the follow-up (p<0.001). Multivariate analysis revealed that BLVRA basal expression in PBL was an independent predictor for sustained virological response (OR 15; 95% CI 1.05-214.2; P = 0.046). HMOX1/2 expression did not have any effect on the treatment outcome.

Conclusion: Our results suggest that patients with chronic HCV infection significantly upregulate BLVRA expression in PBL. The lack of BLVRA overexpression is associated with non-responsiveness to standard antiviral therapy; whereas, HMOX1/2 does not seem to have any predictive potential.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Heme catabolic pathway.
Figure 1. Heme catabolic pathway.
Heme oxygenase, the rate limiting enzyme in the heme catabolic pathway, catalyzes oxidative degradation of heme to form equimolar amounts of bioactive products carbon monoxide, iron and biliverdin, which is subsequently reduced to bilirubin by biliverdin reductase.
Figure 2. ROC curve of BLVRA expression…
Figure 2. ROC curve of BLVRA expression in peripheral blood leukocytes of HCV infected patients.
AUC, area under the ROC curve; ROC, Receiver Operating Charasteristic.
Figure 3. BLVRA expression in peripheral blood…
Figure 3. BLVRA expression in peripheral blood leukocytes of responders and non-SVR patients during standard antiviral therapy.
BLVRA expression was measured the day before treatment initiation (0), and 12, 24, 36 and 48 weeks after start of the standard treatment. Data represent means and standard deviations for triplicate determinations. P-values calculated between responders and non-SVR patients. BLVRA, biliverdin reductase A; PBL, peripheral blood leukocytes; SVR = responders, NVR = non-SVR patients.

References

    1. Seeff LB (1997) Natural history of hepatitis C. Hepatology. 26: 21S–28S.
    1. World Health Organization: Hepatitis C. World Health Organization Fact sheet 164 (Revised June 2011). Available: . Accessed 2012 Feb 4.
    1. Nemecek V, Castkova J, Fritz P, Linhartova A, Svandova E, et al. (2003) The 2001 serological survey in the Czech Republic-viral hepatitis. Cent Eur J Public Health 11 Suppl: S54–61
    1. Nemecek V, Strunecky O (2009) Genotypic heterogeneity of hepatitis C virus (HCV) from blood donors in the Czech Republic. Epidemiol Mikrobiol Imunol 58: 63–72.
    1. Wang JT, Sheu JC, Lin JT, Wang TH, Chen DS (1992) Detection of replicative form of hepatitis C virus RNA in peripheral blood mononuclear cells. J Infect Dis 166: 1167–1169.
    1. Peterhans E (1997) Reactive oxygen species and nitric oxide in viral diseases. Biol Trace Elem Res 56: 107–116.
    1. Okuda M, Li K, Beard MR, Showalter LA, Scholle F, et al. (2002) Mitochondrial injury, oxidative stress, and antioxidant gene expression are induced by hepatitis C virus core protein. Gastroenterology 122: 366–375.
    1. Abdalla MY, Britigan BE, Wen F, Icardi M, McCormick ML, et al. (2004) Down-regulation of heme oxygenase-1 by hepatitis C virus infection in vivo and by the in vitro expression of hepatitis C core protein. J Infect Dis 190: 1109–1118.
    1. Sass G, Seyfried S, M PS, Yamashita K, Kaczmarek E, et al. (2004) Cooperative effect of biliverdin and carbon monoxide on survival of mice in immune-mediated liver injury. Hepatology 40: 1128–1135.
    1. Verma A, Hirsch DJ, Glatt CE, Ronnett GV, Snyder SH (1993) Carbon monoxide: a putative neural messenger. Science 259: 381–384.
    1. Vitek L, Schwertner HA (2007) The heme catabolic pathway and its protective effects on oxidative stress-mediated diseases. Adv Clin Chem 43: 1–57.
    1. Maines MD, Panahian N (2001) The heme oxygenase system and cellular defense mechanisms. Do HO-1 and HO-2 have different functions? Adv Exp Med Biol 502: 249–272.
    1. Willis D, Moore AR, Frederick R, Willoughby DA (1996) Heme oxygenase: a novel target for the modulation of the inflammatory response. Nat Med 2: 87–90.
    1. Li L, Grenard P, Nhieu JT, Julien B, Mallat A, et al. (2003) Heme oxygenase-1 is an antifibrogenic protein in human hepatic myofibroblasts. Gastroenterology 125: 460–469.
    1. Sass G, Soares MC, Yamashita K, Seyfried S, Zimmermann WH, et al. (2003) Heme oxygenase-1 and its reaction product, carbon monoxide, prevent inflammation-related apoptotic liver damage in mice. Hepatology 38: 909–918.
    1. Zhu Z, Wilson AT, Mathahs MM, Wen F, Brown KE, et al. (2008) Heme oxygenase-1 suppresses hepatitis C virus replication and increases resistance of hepatocytes to oxidant injury. Hepatology 48: 1430–1439.
    1. Ghaziani T, Shan Y, Lambrecht RW, Donohue SE, Pietschmann T, et al. (2006) HCV proteins increase expression of heme oxygenase-1 (HO-1) and decrease expression of Bach1 in human hepatoma cells. J Hepatol 45: 5–12.
    1. Baranano DE, Rao M, Ferris CD, Snyder SH (2002) Biliverdin reductase: a major physiologic cytoprotectant. Proc Natl Acad Sci U S A 99: 16093–16098.
    1. Miralem T, Hu Z, Torno MD, Lelli KM, Maines MD (2005) Small interference RNA-mediated gene silencing of human biliverdin reductase, but not that of heme oxygenase-1, attenuates arsenite-mediated induction of the oxygenase and increases apoptosis in 293A kidney cells. J Biol Chem 280: 17084–17092.
    1. Florczyk UM, Jozkowicz A, Dulak J (2008) Biliverdin reductase: new features of an old enzyme and its potential therapeutic significance. Pharmacol Rep 60: 38–48.
    1. Ahmad Z, Salim M, Maines MD (2002) Human biliverdin reductase is a leucine zipper-like DNA-binding protein and functions in transcriptional activation of heme oxygenase-1 by oxidative stress. J Biol Chem 277: 9226–9232.
    1. Kravets A, Hu Z, Miralem T, Torno MD, Maines MD (2004) Biliverdin reductase, a novel regulator for induction of activating transcription factor-2 and heme oxygenase-1. J Biol Chem 279: 19916–19923.
    1. Ding B, Gibbs PE, Brookes PS, Maines MD (2011) The coordinated increased expression of biliverdin reductase and heme oxygenase-2 promotes cardiomyocyte survival: a reductase-based peptide counters beta-adrenergic receptor ligand-mediated cardiac dysfunction. FASEB J 25: 301–313.
    1. Yamaguchi T, Komoda Y, Nakajima H (1994) Biliverdin-IX alpha reductase and biliverdin-IX beta reductase from human liver. Purification and characterization. J Biol Chem 269: 24343–24348.
    1. Maines MD (2010) Potential application of biliverdin reductase and its fragments to modulate insulin/IGF-1/MAPK/PI3-K signaling pathways in therapeutic settings. Curr Drug Targets 11: 1586–1594.
    1. Lehmann E, El-Tantawy WH, Ocker M, Bartenschlager R, Lohmann V, et al. (2010) The heme oxygenase 1 product biliverdin interferes with hepatitis C virus replication by increasing antiviral interferon response. Hepatology 51: 398–404.
    1. Huang C, Chen H, Cassidy W, Howell CD (2008) Peripheral blood gene expression profile associated with sustained virologic response after peginterferon plus ribavirin therapy for chronic hepatitis-C genotype 1. J Natl Med Assoc 100: 1425–1433.
    1. Ghany MG, Strader DB, Thomas DL, Seeff LB (2009) Diagnosis, management, and treatment of hepatitis C: an update. Hepatology 49: 1335–1374.
    1. Ishak K, Baptista A, Bianchi L, Callea F, De Groote J, et al. (1995) Histological grading and staging of chronic hepatitis. J Hepatol 22: 696–699.
    1. Carriere M, Pene V, Breiman A, Conti F, Chouzenoux S, et al. (2007) A novel, sensitive, and specific RT-PCR technique for quantitation of hepatitis C virus replication. J Med Virol 79: 155–160.
    1. Vreman HJ, Stevenson DK (1988) Detection of heme oxygenase activity by measurement of CO. In: Maines MD, Costa LG, Reed DJ, Sassa S, Sipes IG. Current Protocols in Toxicology. New York: John Wiley&Sons. 9.2.1–9.2.10.
    1. Zhu Z, Wilson AT, Luxon BA, Brown KE, Mathahs MM, et al. (2010) Biliverdin inhibits hepatitis C virus nonstructural 3/4A protease activity: mechanism for the antiviral effects of heme oxygenase? Hepatology 52: 1897–1905.
    1. Wen F, Brown KE, Britigan BE, Schmidt WN (2008) Hepatitis C core protein inhibits induction of heme oxygenase-1 and sensitizes hepatocytes to cytotoxicity. Cell Biol Toxicol 24: 175–188.
    1. Ge D, Fellay J, Thompson AJ, Simon JS, Shianna KV, et al. (2009) Genetic variation in IL28B predicts hepatitis C treatment-induced viral clearance. Nature 461: 399–401.
    1. Suppiah V, Moldovan M, Ahlenstiel G, Berg T, Weltman M, et al. (2009) IL28B is associated with response to chronic hepatitis C interferon-alpha and ribavirin therapy. Nat Genet 41: 1100–1104.
    1. Thompson AJ, Muir AJ, Sulkowski MS, Ge D, Fellay J, et al. (2010) Interleukin-28B polymorphism improves viral kinetics and is the strongest pretreatment predictor of sustained virologic response in genotype 1 hepatitis C virus. Gastroenterology 139: 120–129.
    1. Frankova S, Bartakova R, Hejda V, Urbanek P, Husova L, et al. (2012) IL28 rs 12979860 CC genotype slows progression of liver disease in HCV infected patients. Gastroenterologie a Hepatologie 66: S17.
    1. Lanford RE, Guerra B, Lee H, Chavez D, Brasky KM, et al. (2006) Genomic response to interferon-alpha in chimpanzees: implications of rapid downregulation for hepatitis C kinetics. Hepatology 43: 961–972.
    1. Waddell SJ, Popper SJ, Rubins KH, Griffiths MJ, Brown PO, et al. (2010) Dissecting interferon-induced transcriptional programs in human peripheral blood cells. PLoS One 5: e9753.
    1. He XS, Ji X, Hale MB, Cheung R, Ahmed A, et al. (2006) Global transcriptional response to interferon is a determinant of HCV treatment outcome and is modified by race. Hepatology 44: 352–359.

Source: PubMed

Patrocinadores y Colaboradores

Condiciones médicas

Intervenciones de drogas

3
Suscribir