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CIRCULATING HBV RNA CORRELATES WITH INTRAHEPATIC
COVALENTLY CLOSED CIRCULAR DNA (CCCDNA) LEVELS
AND ACTIVITY IN UNTREATED CHRONIC HEPATITIS B (CHB)
PATIENTS
Barbara Testoni1, Caroline Scholtès1,2, Marie-Laure
Plissonnier3, Francoise Berby1, Floriana Facchetti4, François
Villeret5, Alessandro Loglio6, Beth Scott7, Ling Wang7, Aaron
Hamilton7, Marintha Heil7, Pietro Lampertico8,9, Massimo
Levrero1,10,11 and Fabien Zoulim1,11, (1)Cancer Research
Center of Lyon (CRCL), Umr Inserm U1052-Cnrs 5286 Mixte
Clb, Lyon1 University, France, (2)Department of Infectious
Diseases, Croix-Rousse Hospital, Hospices Civils De Lyon,
France, (3)Cancer Research Center of Lyon (CRCL), Umr
Inserm U1052-Cnrs 5286 Mixte Clb, Lyon-1 University, France,
(4)Division of Gastroenterology and Hepatology - CRC “a. M.
and a. Migliavacca” Center for Liver Disease, Foundation
Irccs Ca’ Granda Ospedale Maggiore Policlinico, Italy, (5)
Hepatology Department, Hospices Civils De Lyon, France,
(6)Division of Gastroenterology and Hepatology - CRC AM
and a Migliavacca” Center for Liver Diseases, Foundation
Irccs Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy,
(7)Department of Infectious Diseases, Roche Molecular
Diagnostics, Inc., CA, (8)Department of Pathophysiology
and Transplantation, University of Milan, Italy, (9)Division
of Gastroenterology and Hepatology - CRC “a. M. and a.
Migliavacca” Center for Liver Disease, Foundation Irccs
Ca’ Granda Ospedale Maggiore Policlinico - Milan, Italy,
(10)Department of Medicine Sciac, University of Rome La
Sapienza, Italy, (11)Department of Hepatology, Croix-Rousse
Hospital, Hospices Civils De Lyon, France
Background: A comprehensive analysis of Hepatitis B virus
(HBV) RNA in blood circulation (cirB-RNA) in the different
chronic hepatitis B (CHB) phases and its correlation with
intrahepatic viral markers and HBcrAg, the other emerging
biomarker of cccDNA transcription, is still lacking Methods:
122 untreated CHB patients with paired liver biopsy and serum
sample, 32 HBeAg(+) chronic hepatitis (CH), 29 HBeAgchronic
infection (CI) and 61 HBeAg(-) CH, were analyzed for
serum HBV DNA, quantitative (q)HBsAg, HBcrAg and alanine
aminotransferase (ALT) levels Liver cccDNA and 3 5Kb RNA
were assessed by qPCR and RT-qPCR, respectively, and
cccDNA transcriptional activity was calculated as 3 5Kb RNA/
cccDNA ratio Liver histology scores were also available cirBRNA
was quantified by the Roche HBV RNA assay for use
on the cobas® 6800/8800 Systems (LLOQ 10 cp/ml; linearity
range 10 to 107cp/ml; LLOD ~3cp/mL) Results: While all
HBeAg(+) CH were cirB-RNA(+), 57% of HBeAg(-) CH and
only 14% of HBeAg(-) CI patients had quantifiable cirB-RNA.
cirB-RNA was higher in HBeAg(+) vs HBeAg(-) patients
(median of 5 4 vs 2 3 log10 copies/ml, p <0 0001), in patients
with necroinflammatory activity score >1 (3 vs 1.4 log copies/
ml, p=0.001) and in those with ALT>2N (5.1 vs 2 log copies/
ml, p<0 0001) The 39 cirB-RNA(-) patients (23 HBeAg(-) CI
and 16 HBeAg(-) CH) had lower cccDNA (median 0 06 vs
0 3 copies/cell, p<0 0001), 3 5Kb RNA (median 0 03 vs 10 3,
p<0 0001) and 3 5Kb RNA/cccDNA (0 79 vs 8 9, p=0 002) as
compared to the cirB-RNA(+) ones. No significant difference
was found in qHBsAg levels, while both HBcrAg (median
5 4 vs 2 6 LogU/ml, p<0 0001) and serum HBV DNA (6 2
vs 2.7 LogIU/ml, p<0.0001) were significantly higher in cirBRNA(+)
patients. In HBeAg(-) patients, cirB-RNA significantly
correlated with serum HBV DNA (R=0 81, p<0 0001), HBcrAg
(R=0 73, p<0 0001), intrahepatic 3 5Kb RNA (R=0 77,
p<0 001) and cccDNA transcriptional activity (R=0 78,
p<0 0001), but not with HBsAg (R=0 16, p=ns) and cccDNA
levels (R=0 11, p=ns) In HBeAg(-) CH group, a cirB-RNA
cut-off >3 log10 copies/ml identified a population of patients
with higher intrahepatic 3 5Kb RNA/cccDNA ratio (median of
2 8 vs 0 7 log10 in the overall HBeAg(-) CH group) and more
advanced liver fibrosis, but without difference in HBsAg levels.
Conclusion: Our results support the notion that cirB-RNA
detected by Roche HBV RNA assay reflects the transcriptional
activity of intrahepatic cccDNA This work is supported by the
French National Research Agency Investissements d’Avenir
program (CirB-RNA project – ANR-17-RHUS-0003)作者: StephenW 时间: 2020-10-22 15:13