AASLD2020[722]HBV RNA與肝內循環相關

StephenW

722
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)

StephenW

722
HBV RNA與肝內循環相關
共封閉圓形DNA（CCCDNA）水平
和未治療的慢性乙型肝炎（CHB）的活性
耐心
芭芭拉·泰斯托尼（Barbara Testoni）1，卡羅琳·舒爾特（CarolineScholtès）1,2，瑪麗·勞雷（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和Fabien Zoulim1,11，（1）癌症研究
里昂市中心（CRCL），Umr Inserm U1052-Cnrs 5286 Mixte
法國里昂1大學（Clb），傳染病學系（2）
疾病，Croix-Rousse醫院，Hospices Civils De Lyon，
法國，（3）里昂癌症研究中心（CRCL）
Inserm U1052-Cnrs 5286 Mixte Clb，法國里昂1大學，
（4）胃腸病學和肝病學-CRC“ a。 M.
和一個。 Migliavacca”基金會肝病中心
意大利Irccs Ca’Granda Ospedale Maggiore Policlinico，（5）
法國里昂平民醫院肝病科，
（6）消化內科-CRC AM
和基金會的Migliavacca肝病中心
Irccs Ca’Granda Ospedale Maggiore Policlinico，米蘭，意大利，
（7）羅氏分子醫院傳染病學系
加利福尼亞州Diagnostics，Inc.（8）病理生理學系
和移植，意大利米蘭大學，（9）
胃腸病學和肝病學-CRC“ a。 M.和
Migliavacca”基金會Irccs肝病中心
Ca’Granda Ospedale Maggiore Policlinico-意大利米蘭，
（10）羅馬大學西亞夏醫學院
意大利薩皮恩扎（11）Croix-Rousse肝病科
法國里昂市民醫院醫院
背景：乙肝病毒的綜合分析
（HBV）血液循環中的RNA（cirB-RNA）不同
慢性乙型肝炎（CHB）階段及其與疾病的相關性
肝內病毒標誌物和HBcrAg，另一個正在出現
仍缺乏cccDNA轉錄的生物標誌物方法：
122例未經治療的CHB患者與肝活檢和血清配對
樣本，32 HBeAg（+）慢性肝炎（CH），29 HBeAg慢性
分析感染（CI）和61 HBeAg（-）CH
血清HBV DNA，定量（q）HBsAg，HBcrAg和丙氨酸
氨基轉移酶（ALT）水平肝cccDNA和3個5Kb RNA
分別通過qPCR和RT-qPCR進行評估，並且
cccDNA轉錄活性計算為3 5Kb RNA /
cccDNA比率肝組織學評分也可用cirBRNA
通過Roche HBV RNA測定法定量使用
在cobas®6800/8800系統上（LLOQ 10 cp / ml;線性
範圍10至107cp / ml; LLOD〜3cp / mL）結果：雖然全部
HBeAg（+）CH為cirB-RNA（+），HBeAg（-）CH為57％，
HBeAg（-）CI患者中只有14％具有可定量的cirB-RNA。
HbeAg（+）患者中的cirB-RNA高於HBeAg（-）患者
（患者中位數為5 4 vs 2 3 log10個拷貝/ ml，p <0 0001）
壞死性炎症評分> 1（3 vs 1.4日誌副本/
ml，p = 0.001）和ALT> 2N（5.1 vs 2對數副本/
ml，p <0 0001）39例cirB-RNA（-）患者（23 HBeAg（-）CI
和16 HBeAg（-）CH）的cccDNA較低（中位數0 06 vs
0 3拷貝/細胞，p <0 0001），3 5Kb RNA（中值0 03 vs 10 3，
p <0 0001）和3個5Kb RNA / cccDNA（0 79 vs 8 9，p = 0 002）
與cirB-RNA（+）相比。無明顯差異
被發現在qHBsAg水平，而兩個HBcrAg（中間
5 4 vs 2 6 LogU / ml，p <00001）和血清HBV DNA（6 2
vs 2.7 LogIU / ml，p <0.0001）在cirBRNA（+）中顯著更高
耐心。在HBeAg（-）患者中，cirB-RNA顯著
與血清HBV DNA相關（R = 0 81，p <0 0001），HBcrAg
（R = 0 73，p <0 0001），肝內3 5Kb RNA（R = 0 77，
p <0 001）和cccDNA轉錄活性（R = 0 78，
p <0 0001），但不包含HBsAg（R = 0 16，p = ns）和cccDNA
水平（R = 0 11，p = ns）在HBeAg（-）CH組中，cirB-RNA
臨界值> 3 log10拷貝/ ml確定了患者人群
肝內3 5Kb RNA / cccDNA比值更高（
整個HBeAg（-）CH組的2 8 vs 0 7 log10
晚期肝纖維化，但HBsAg水平無差異。
結論：我們的結果支持cirB-RNA的觀點
羅氏HBV RNA檢測法檢測到了轉錄
肝內cccDNA的活性
法國國家研究機構大道（Investissements d’Avenir）
程序（CirB-RNA項目– ANR-17-RHUS-0003）