TY - JOUR
T1 - Control of APOBEC3B induction and cccDNA decay by NF-κB and miR-138-5p
AU - Faure-Dupuy, Suzanne
AU - Riedl, Tobias
AU - Rolland, Maude
AU - Hizir, Zoheir
AU - Reisinger, Florian
AU - Neuhaus, Katharina
AU - Schuehle, Svenja
AU - Remouchamps, Caroline
AU - Gillet, Nicolas
AU - Schönung, Maximilian
AU - Stadler, Mira
AU - Wettengel, Jochen
AU - Barnault, Romain
AU - Parent, Romain
AU - Schuster, Linda Christina
AU - Farhat, Rayan
AU - Prokosch, Sandra
AU - Leuchtenberger, Corinna
AU - Öllinger, Rupert
AU - Engleitner, Thomas
AU - Rippe, Karsten
AU - Rad, Roland
AU - Unger, Kristian
AU - Tscharahganeh, Darjus
AU - Lipka, Daniel B
AU - Protzer, Ulrike
AU - Durantel, David
AU - Lucifora, Julie
AU - Dejardin, Emmanuel
AU - Heikenwälder, Mathias
N1 - Funding Information:
M.H. was supported by an ERC Consolidator grant (HepatoMetaboPath), SFBTR179 Project-ID 272983813, SFB/TR 209 Project-ID 314905040, SFBTR1335 Project-ID 360372040, the Wilhelm Sander-Stiftung , a Horizon 2020 grant (Hepcar), Research Foundation Flanders (FWO) under grant 30826052 (EOS Convention MODEL-IDI), Deutsche Krebshilfe projects 70113166 and 70113167 , German-Israeli Cooperation in Cancer Research (DKFZ-MOST) and the Helmholtz-Gemeinschaft , Zukunftsthema “Immunology and Inflammation” (ZT-0027), and the Rainer Hoenig Stiftung . E.D. and M.H. were supported by the FNRS/FWO under EOS project no. 30826052. E.D. received financial support from the F.R.S.-FNRS (CDR-J.0049.20) and the Fondation Léon Fredericq of the University of Liege . Z.H. was supported a Grant Télévie , Belgium. M.H., E.D., D.D., J.L., and M.R. were supported by an FP7-Infect-Era grant and a fellowship from the WBI (Wallonie-Bruxelles International). D.D. and J.L. were supported by INSERM (Institut National de la Santé et de la Recherche Médicale; salaries and core-fundings), ANRS (Agence Nationale de Recherche sur le Sida et les hépatites virales, several grants from study section 12 [CSS12]), and EU-Infect Era “Target HDV” ( ANR 16-IFEC-0005-01 ).
Publisher Copyright:
© 2021 The Authors
PY - 2021/12
Y1 - 2021/12
N2 - Background & Aims: Immune-mediated induction of cytidine deaminase APOBEC3B (A3B) expression leads to HBV covalently closed circular DNA (cccDNA) decay. Here, we aimed to decipher the signalling pathway(s) and regulatory mechanism(s) involved in A3B induction and related HBV control.Methods: Differentiated HepaRG cells (dHepaRG) knocked-down for NF-κB signalling components, transfected with siRNA or micro RNAs (miRNA), and primary human hepatocytes ± HBV or HBVΔX or HBV-RFP, were treated with lymphotoxin beta receptor (LTβR)-agonist (BS1). The biological outcomes were analysed by reverse transcriptase-qPCR, immunoblotting, luciferase activity, chromatin immune precipitation, electrophoretic mobility-shift assay, targeted-bisulfite-, miRNA-, RNA-, genome-sequencing, and mass-spectrometry.Results: We found that canonical and non-canonical NF-κB signalling pathways are mandatory for A3B induction and anti-HBV effects. The degree of immune-mediated A3B production is independent of A3B promoter demethylation but is controlled post-transcriptionally by the miRNA 138-5p expression (hsa-miR-138-5p), promoting A3B mRNA decay. Hsa-miR-138-5p over-expression reduced A3B levels and its antiviral effects. Of note, established infection inhibited BS1-induced A3B expression through epigenetic modulation of A3B promoter. Twelve days of treatment with a LTβR-specific agonist BS1 is sufficient to reduce the cccDNA pool by 80% without inducing significant damages to a subset of cancer-related host genes. Interestingly, the A3B-mediated effect on HBV is independent of the transcriptional activity of cccDNA as well as on rcDNA synthesis.Conclusions: Altogether, A3B represents the only described enzyme to target both transcriptionally active and inactive cccDNA. Thus, inhibiting hsa-miR-138-5p expression should be considered in the combinatorial design of new therapies against HBV, especially in the context of immune-mediated A3B induction.Lay summary: Immune-mediated induction of cytidine deaminase APOBEC3B is transcriptionally regulated by NF-κB signalling and post-transcriptionally downregulated by hsa-miR-138-5p expression, leading to cccDNA decay. Timely controlled APOBEC3B-mediated cccDNA decay occurs independently of cccDNA transcriptional activity and without damage to a subset of cancer-related genes. Thus, APOBEC3B-mediated cccDNA decay could offer an efficient therapeutic alternative to target hepatitis B virus chronic infection.
AB - Background & Aims: Immune-mediated induction of cytidine deaminase APOBEC3B (A3B) expression leads to HBV covalently closed circular DNA (cccDNA) decay. Here, we aimed to decipher the signalling pathway(s) and regulatory mechanism(s) involved in A3B induction and related HBV control.Methods: Differentiated HepaRG cells (dHepaRG) knocked-down for NF-κB signalling components, transfected with siRNA or micro RNAs (miRNA), and primary human hepatocytes ± HBV or HBVΔX or HBV-RFP, were treated with lymphotoxin beta receptor (LTβR)-agonist (BS1). The biological outcomes were analysed by reverse transcriptase-qPCR, immunoblotting, luciferase activity, chromatin immune precipitation, electrophoretic mobility-shift assay, targeted-bisulfite-, miRNA-, RNA-, genome-sequencing, and mass-spectrometry.Results: We found that canonical and non-canonical NF-κB signalling pathways are mandatory for A3B induction and anti-HBV effects. The degree of immune-mediated A3B production is independent of A3B promoter demethylation but is controlled post-transcriptionally by the miRNA 138-5p expression (hsa-miR-138-5p), promoting A3B mRNA decay. Hsa-miR-138-5p over-expression reduced A3B levels and its antiviral effects. Of note, established infection inhibited BS1-induced A3B expression through epigenetic modulation of A3B promoter. Twelve days of treatment with a LTβR-specific agonist BS1 is sufficient to reduce the cccDNA pool by 80% without inducing significant damages to a subset of cancer-related host genes. Interestingly, the A3B-mediated effect on HBV is independent of the transcriptional activity of cccDNA as well as on rcDNA synthesis.Conclusions: Altogether, A3B represents the only described enzyme to target both transcriptionally active and inactive cccDNA. Thus, inhibiting hsa-miR-138-5p expression should be considered in the combinatorial design of new therapies against HBV, especially in the context of immune-mediated A3B induction.Lay summary: Immune-mediated induction of cytidine deaminase APOBEC3B is transcriptionally regulated by NF-κB signalling and post-transcriptionally downregulated by hsa-miR-138-5p expression, leading to cccDNA decay. Timely controlled APOBEC3B-mediated cccDNA decay occurs independently of cccDNA transcriptional activity and without damage to a subset of cancer-related genes. Thus, APOBEC3B-mediated cccDNA decay could offer an efficient therapeutic alternative to target hepatitis B virus chronic infection.
KW - APOBEC3B
KW - cccDNA
KW - HBx
KW - Hepatitis B virus
KW - miRNA
KW - NF-κB
UR - http://www.scopus.com/inward/record.url?scp=85122807390&partnerID=8YFLogxK
U2 - 10.1016/j.jhepr.2021.100354
DO - 10.1016/j.jhepr.2021.100354
M3 - Article
C2 - 34704004
SN - 2589-5559
VL - 3
SP - 100354
JO - JHEP reports : innovation in hepatology
JF - JHEP reports : innovation in hepatology
IS - 6
M1 - 100354
ER -