Hepatitis B virus genetic diversity has minimal impact on sensitivity of the viral ribonuclease H to inhibitors.
Hepatitis B virus (HBV) causes hepatitis, cirrhosis, liver failure, and liver cancer, but the current therapies that employ either nucelos(t)ide analogs or (pegylated)interferon α do not clear the infection in the large majority of patients. Inhibitors of the HBV ribonuclease H (RNaseH) that are being developed with the goal of producing anti-HBV drugs are promising candidates for use in combination with the nucleos(t)ide analogs to improve therapeutic efficacy. HBV is genetically very diverse, with at least 8 genotypes that differ by ≥8% at the sequence level. This diversity is reflected in the viral RNaseH enzyme, raising the possibility that divergent HBV genotypes or isolates may have varying sensitivity to RNaseH inhibitors. To evaluate this possibility, we expressed and purified 18 patient-derived RNaseHs from genotypes B, C, and D. Basal RNaseH activity and sensitivity to three novel RNaseH inhibitors from three different chemotypes were assessed. We also evaluated four consensus HBV RNaseHs to determine if such sequences would be suitable for use in antiviral drug screening. The patient-derived enzymes varied by over 10-fold in their basal RNaseH activities, but they were equivalently sensitive to each of the three inhibitors. Similarly, all four consensus HBV RNaseH enzymes were active and were equally sensitive to an RNaseH inhibitor. These data indicate that a wide range of RNaseH sequences would be suitable for use in antiviral drug screening, and that genotype- or isolate-specific genetic variations are unlikely to present a barrier during antiviral drug development against the HBV RNaseH.
Department of Molecular Microbiology and Immunology and Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA; Department of Gastroenterology, The Second Affiliated Hospital, Zhengzhou University, Zhengzhou, Henan, People's Republic of China.
Gaofeng
G
Lu
Department of Molecular Microbiology and Immunology and Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA.
Juan Antonio
JA
Villa
Department of Molecular Microbiology and Immunology and Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA.
Maureen J
MJ
Donlin
Department of Molecular Microbiology and Immunology and Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA.
Tiffany C
TC
Edwards
Department of Molecular Microbiology and Immunology and Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA.
Xiaohong
X
Cheng
Center for World Health and Medicine, Saint Louis University School of Medicine, St. Louis, MO, USA.
Richard F
RF
Heier
Center for World Health and Medicine, Saint Louis University School of Medicine, St. Louis, MO, USA.
Marvin J
MJ
Meyers
Department of Molecular Microbiology and Immunology and Saint Louis University Liver Center, Saint Louis University School of Medicine, St. Louis, MO, USA. Electronic address: tavisje@slu.edu.
John E
JE
Tavis
D000998:Antiviral Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D014644:Genetic Variation; D005838:Genotype; D006515:Hepatitis B virus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000188:drug therapy; D006801:Humans; D016914:Ribonuclease H / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D016914:Ribonuclease H
DNA virus; Genetic diversity; Hepatitis B virus; Ribonuclease H
PLoS Pathog. 2015 Jun 18;11(6):e1004854
26087286
Cell. 2005 Jul 1;121(7):1005-16
15989951
J Virol. 1996 Jul;70(7):4269-74
8676448
Vaccine. 2005 Mar 31;23(19):2409-23
15752827
Nat Rev Gastroenterol Hepatol. 2011 May;8(5):275-84
21423260
Antimicrob Agents Chemother. 2015 Feb;59(2):1070-9
25451058
Antivir Chem Chemother. 2004 Nov;15(6):299-305
15646643
Antiviral Res. 2013 Apr;98(1):27-34
23391846
Hepatology. 2010 Nov;52(5):1611-20
20931556
N Engl J Med. 2004 Mar 11;350(11):1118-29
15014185
Nucleic Acids Res. 2003 Dec 1;31(23):6852-9
14627818
J Viral Hepat. 2011 Jun;18(6):415-23
20626626
Expert Rev Anti Infect Ther. 2013 Aug;11(8):755-7
23977931
World J Gastroenterol. 2013 Dec 21;19(47):8887-94
24379612
Bioorg Med Chem Lett. 2010 Nov 15;20(22):6754-7
20869872
Antiviral Res. 2016 Aug;132:186-95
27321664
J Virol. 2006 Nov;80(21):10307-14
17041211
Expert Opin Emerg Drugs. 2011 Dec;16(4):713-29
22195605
J Viral Hepat. 2011 May;18(5):349-57
20529202
PLoS Pathog. 2013 Jan;9(1):e1003125
23349632
EMBO Rep. 2009 Feb;10(2):144-51
19165139
Hepatol Res. 2010 Jan;40(1):14-30
20156297
J Virol. 1996 Sep;70(9):6151-6
8709240
Hepatology. 2010 Jan;51(1):73-80
19998272
Curr Gastroenterol Rep. 2012 Feb;14(1):37-46
22105466
Bioinformatics. 2009 Jan 1;25(1):126-7
18984599
Proteins. 2013 Nov;81(11):1857-61
23852738
EMBO J. 2006 May 3;25(9):1924-33
16601679
J Virol. 1994 Aug;68(8):5232-8
8035519
Antiviral Res. 2013 Sep;99(3):221-9
23796982
Antiviral Res. 2014 Aug;108:48-55
24858512
BMC Immunol. 2016 Jun 16;17(1):17
27307005
PLoS One. 2015 May 05;10(5):e0125555
25942596
Ann Intern Med. 2009 Jan 20;150(2):104-10
19124811
Structure. 1995 Feb 15;3(2):131-4
7735828
Epidemiol Rev. 2006;28:112-25
16754644
Science. 1994 Dec 23;266(5193):1981-6
7801124
Gastroenterology. 2010 Oct;139(4):1218-29
20600036
Antiviral Res. 2015 Jun;118:132-8
25862291
J Viral Hepat. 2004 Mar;11(2):97-107
14996343
PLoS One. 2015 Sep 25;10(9):e0136074
26406309
N Engl J Med. 2008 Dec 4;359(23):2442-55
19052126