AASLD2016[599]减少肝内cccDNA水平允许调节 HBV复制后停止治疗 人

StephenW

599
Reduction of intrahepatic cccDNA levels permits to regulate
HBV replication after cessation of the treatment in
human hepatocyte chimeric mice
Takuro Uchida1, Michio Imamura1, Hiromi Kan1, Nobuhiko
Hiraga1, Masataka Tsuge1, Hiromi Abe-Chayama1, Hiroshi
Aikata1, C. Nelson Hayes1, Yuji Ishida2, Chise Tateno2, Kazuaki
Chayama1; 1Department of Gastroenterology and Metabolism,
Hiroshima University, Hiroshima, Japan; 2PhoenixBio Co., Ltd.,
Higashihiroshima, Japan
Background & aims: Hepatitis B virus (HBV) infection is the most
common chronic viral infection in the world. Nucleot(s)ide analogues
and peg-interferon (PEG-IFN) are treatment options for
patients with chronic HBV infection. However, cure is rare due
to the persistence of viral DNA in hepatocytes in the form of stable
covalently closed circular DNA (cccDNA). In this study, we
investigated the effect of the extreme reduction of intrahepatic
cccDNA levels on HBV replication after cessation of the treatment
using HBV-infected humanized mouse model. Methods:
Human hepatocyte transplanted uPA-SCID mice were infected
with HBV, then administered with 2 mg/kg of entecavir (ETV)
daily or twice a week with 30 μg/kg of PEG-IFN, or a combination
of these drugs, for 12 weeks. ETV was administered
orally once per day and PEG-IFNα-2a was administered subcutaneously
twice a week. Serum HBV DNA, hepatitis B surface
antigen (HBsAg), hepatitis B e antigen (HBeAg), and hepatitis
B core-related antigen (HBcrAg) were measured. We extracted
RNA and DNA from mouse liver samples and measured intrahepatic
HBV DNA, cccDNA, and viral protein transcription
levels. Results: HBV-infected mice were treated with ETV alone,
PEG-IFN alone, or ETV plus PEG-IFN for 12 weeks. Treatment
with ETV alone and PEG-IFN alone resulted in 4.0 and 1.7
log reductions in serum HBV DNA levels, respectively. HBV
DNA levels in the mouse treated with combination treatment
decreased below detectable levels (4.4 log copies/mL). HBsAg
and HBeAg values significantly decreased in mice treated with
either PEG-IFN alone or in combination with ETV. ETV and
PEG-IFN combination resulted in a significant reduction of intrahepatic
HBV DNA, cccDNA, and transcription of viral proteins.
To reduce intrahepatic cccDNA levels, mice were treated
with 20 mg/kg of entacavir plus 300 μg/kg of PEG-IFN for
6 weeks. High doses of entacavir plus PEG-IFN combination
treatment resulted in a rapid reduction of mouse serum HBV
DNA, HBsAg and HBcrAg levels. Six weeks of combination
therapy reduced mouse intrahepatic HBV DNA and cccDNA
levels to 0.1 and 0.01 copies/cell, respectively. After cessation
of the combination treatment, serum HBsAg, HBcrAg, intrahepatic
HBV DNA, and cccDNA levels rebounded in mice and
serum HBV DNA reappeared. In contrast, these HBV markers
remained at low levels in mice with persistently negative serum
HBV DNA until 12 weeks after cessation of the therapy. Conclusion:
Extreme reduction of intrahepatic cccDNA levels by
anti-HBV treatment could permit suppression of HBV replication
even after cessation of the treatment.
Disclosures:
Michio Imamura - Grant/Research Support: Bristol-Meyers Squibb; Speaking and
Teaching: Bristol-Meyers Squibb
Yuji Ishida - Employment: PhoenixBio Co., Ltd.
Chise Tateno - Board Membership: PhoenixBio Co., Ltd.
Kazuaki Chayama - Advisory Committees or Review Panels: Mitsubishi Tanabe,
Taisho Toyama; Consulting: AbbVie; Grant/Research Support: Ajinomoto, Abb-
Vie, Aska, Asstellas, Aska, Bristol Squibb, Daiichi Sankyo, Dainippon Sumitomo,
Daiichi Sankyo, Eisai, GlaxoSmithKline, Mitsubishi Tanabe, Nippon Kayaku,
Otsuka, Sogo Rinsho Médéfi, Taiho, Takeda, Toray, Torii, Tsumura, Zeria;
Speaking and Teaching: Abbott, AbbVie, Ajinomoto, Astellas, AstraZeneca,
Bayer, Bristol Squibb, Chugai, Dainippon Sumitomo, Eidia, Eisai, Gilead,
GlaxoSmithKline, JIMRO, Johnson & Johnson, Mitsubishi Tanabe
The following people have nothing to disclose: Takuro Uchida, Hiromi Kan,
Nobuhiko Hiraga, Masataka Tsuge, Hiromi Abe-Chayama, Hiroshi Aikata, C.
Nelson Hayes

StephenW

AASLD2016 [599]减少肝内cccDNA水平允许调节
HBV复制后停止治疗
人肝细胞嵌合小鼠
减少肝内cccDNA水平允许调节
HBV复制后停止治疗
人肝细胞嵌合小鼠
Takuro Uchida1，Michio Imamura1，Hiromi Kan1，Nobuhiko
Hiraga1，Masataka Tsuge 1，Hiromi Abe-Chayama1，Hiroshi
Aikata1，C.Nelson Hayes1，Yuji Ishida2，Chise Tateno2，Kazuaki
Chayama1;消化内科及代谢，
广岛大学，广岛，日本; 2PhoenixBio有限公司，
Higashihiroshima，Japan
背景和目的：乙型肝炎病毒（HBV）感染是最多的
常见的慢性病毒感染在世界上。核酸类似物
和聚乙二醇干扰素（PEG-IFN）是治疗选择
慢性HBV感染患者。然而，治愈是罕见的
以稳定的形式在肝细胞中的病毒DNA的持久性
共价闭合环状DNA（cccDNA）。在这项研究中，我们
研究了肝内极端减少的影响
cccDNA水平对HBV复制停止治疗后
使用HBV感染的人源化小鼠模型。方法：
移植人肝细胞的uPA-SCID小鼠
与HBV，然后与2毫克/公斤恩替卡韦（ETV）
每天或每周两次，用30μg/ kg的PEG-IFN或其组合
的这些药物，12周。 ETV
口服每天一次，并且皮下施用PEG-IFNα-2a
每周两次。血清HBV DNA，乙型肝炎表面
抗原（HBsAg），乙型肝炎e抗原（HBeAg）和肝炎
B核心相关抗原（HBcrAg）。我们提取
RNA和DNA，并测量肝内
HBV DNA，cccDNA和病毒蛋白转录
水平。结果：HBV感染小鼠单独用ETV治疗，
PEG-IFN单独或ETV加PEG-IFN治疗12周。治疗
单独的ETV和单独的PEG-IFN导致4.0和1.7
log血清HBV DNA水平的降低。 HBV
用联合治疗处理的小鼠中的DNA水平
降低至低于可检测水平（4.4log copies / mL）。 HBsAg
和HBeAg值在用治疗的小鼠中显着降低
单独的PEG-IFN或与ETV组合。 ETV和
PEG-IFN组合导致肝内的显着减少
HBV DNA，cccDNA和病毒蛋白的转录。
为了降低肝内cccDNA水平，处理小鼠
与20mg / kg的恩他卡韦加上300μg/ kg的PEG-IFN
6周。高剂量的恩他卡韦加PEG-IFN组合
治疗导致小鼠血清HBV的快速减少
DNA，HBsAg和HBcrAg水平。六周的组合
治疗减少小鼠肝内HBV DNA和cccDNA
水平分别为0.1和0.01个拷贝/细胞。停止后
的联合治疗，血清HBsAg，HBcrAg，肝内
HBV DNA和cccDNA水平在小鼠中反弹
血清HBV DNA重新出现。相比之下，这些HBV标志物
在具有持续阴性血清的小鼠中保持在低水平
HBV DNA直到停止治疗后12周。结论：
极低的肝内cccDNA水平降低
抗HBV治疗可以允许抑制HBV复制
甚至在停止治疗后。
披露：
Michi Imamura - 资助/研究支持：Bristol-Meyers Squibb;说话和
教学：Bristol-Meyers Squibb
就业：凤凰实业有限公司
Chise Tateno - 董事会成员：PhoenixBio有限公司
Kazuaki Chayama - 咨询委员会或审查小组：Mitsubishi Tanabe，
富山大咨询：AbbVie;资助/研究支持：Ajinomoto，Abb-
Vie，Aska，Asstellas，Aska，Bristol Squibb，Daiichi Sankyo，Dainippon Sumitomo，
Daiichi Sankyo，Eisai，GlaxoSmithKline，Mitsubishi Tanabe，Nippon Kayaku，
Otsuka，Sogo RinshoM√©d√©fi，Taiho，Takeda，Toray，Torii，Tsumura，Zeria;
讲座和教学：Abbott，AbbVie，Ajinomoto，Astellas，AstraZeneca，
Bayer，Bristol Squibb，Chugai，Dainippon Sumitomo，Eidia，Eisai，Gilead，
GlaxoSmithKline，JIMRO，Johnson＆Johnson，Mitsubishi Tanabe
以下人没有什么可披露：Takuro Uchida，Hiromi Kan，
Nobuhiko Hiraga，Masataka Tsuge，Hiromi Abe-Chayama，Hiroshi Aikata，C.
尼尔森·海斯

MP4

http://phoenixbio.co.jp/en/pxb/pxb-mouse.html