Toll样受体7激动剂GS-9620通过I型干扰素依赖性机制诱导HBV的延

StephenW

   J Hepatol. 2017 Dec 13. pii: S0168-8278(17)32489-3. doi: 10.1016/j.jhep.2017.12.007. [Epub ahead of print]
Toll-Like Receptor 7 Agonist GS-9620 Induces Prolonged Inhibition of HBV via a Type I Interferon-Dependent Mechanism.
Niu C1, Li L1, Daffis S1, Lucifora J2, Bonnin M2, Maadadi S2, Salas E1, Chu R1, Ramos H1, Livingston CM1, Beran RK1, Garg AV1, Balsitis S1, Durantel D2, Zoulim F3, Delaney WE 4th1, Fletcher SP4.
Author information

1
    Gilead Sciences, Inc, Foster City, CA, USA.
2
    INSERM 1052, Université Claude Bernard Lyon 1, CNRS 5286, Centre Léon Bérard, Centre de Recherche en Cancérologie de Lyon, Lyon 69003, France.
3
    INSERM 1052, Université Claude Bernard Lyon 1, CNRS 5286, Centre Léon Bérard, Centre de Recherche en Cancérologie de Lyon, Lyon 69003, France; Hospices Civils de Lyon (HCL), 69002 Lyon, France; Institut Universitaire de France (IUF), 75005 Paris, France.
4
    Gilead Sciences, Inc, Foster City, CA, USA. Electronic address: [email protected].

Abstract
BACKGROUND & AIMS:

GS-9620, an oral agonist of toll-like receptor 7 (TLR7), is in clinical development for the treatment of chronic hepatitis B (CHB). GS-9620 was previously shown to induce prolonged suppression of serum viral DNA and antigens in the woodchuck and chimpanzee models of CHB. Here we investigated the molecular mechanisms that contribute to the antiviral response to GS-9620 using in vitro models of hepatitis B virus (HBV) infection.
METHODS:

Cryopreserved primary human hepatocytes (PHH) and differentiated HepaRG (dHepaRG) cells were infected with HBV and treated with GS-9620, conditioned media from human peripheral blood mononuclear cells (PBMCs) treated with GS-9620 (GS-9620 conditioned media; GS-9620-CM), or other innate immune stimuli. The antiviral and transcriptional response to these agents was determined.
RESULTS:

GS-9620 had no antiviral activity in HBV-infected PHH, consistent with low level TLR7 mRNA expression in human hepatocytes. In contrast, GS-9620-CM induced prolonged reduction of HBV DNA, RNA, and antigen levels in PHH and dHepaRG cells via a type I interferon (IFN)-dependent mechanism. GS-9620-CM did not reduce cccDNA levels in either cell type. Transcriptional profiling demonstrated that GS-9620-CM strongly induced various HBV restriction factors - although not APOBEC3A or the Smc5/6 complex - and indicated that established HBV infection does not modulate innate immune sensing or signaling in cryopreserved PHH. GS-9620-CM also induced expression of immunoproteasome subunits and enhanced presentation of an immunodominant viral peptide in HBV-infected PHH.
CONCLUSIONS:

Type I IFN induced by GS-9620 durably suppressed HBV in human hepatocytes without reducing cccDNA levels. Moreover, HBV antigen presentation was enhanced, suggesting additional components of the TLR7-induced immune response played a role in the antiviral response to GS-9620 in animal models of CHB.
LAY SUMMARY:

GS-9620 is a drug in clinical trials for the treatment of chronic hepatitis B virus (HBV) infection. GS-9620 has previously been shown to suppress HBV in various animal models, but the underlying antiviral mechanisms were not completely understood. In this study, we determined that GS-9620 does not directly activate antiviral pathways in human liver cells, but can induce prolonged suppression of HBV via induction of an antiviral cytokine called interferon. However, interferon did not destroy the HBV genome, suggesting that other parts of the immune response (e.g. activation of immune cells that kill infected cells) also play an important role in the antiviral response to GS-9620.

Copyright © 2017 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
KEYWORDS:

APOBEC; GS-9620; Hepatitis B virus; Immunoproteasome; Interferon-alfa; Interferon-stimulated gene; MHC; Smc5/6; TLR7; cccDNA

PMID:
    29247725
DOI:
    10.1016/j.jhep.2017.12.007

StephenW

J Hepatol。 2017 Dec 13。pii：S0168-8278（17）32489-3。 doi：10.1016 / j.jhep.2017.12.007。 [电子版提前打印]
Toll样受体7激动剂GS-9620通过I型干扰素依赖性机制诱导HBV的延长的抑制。
Ni C1，Li L1，Daffis S1，Lucifora J2，Bonnin M2，Maadadi S2，Salas E1，Chu R1，Ramos H1，Livingston CM1，Beran RK1，Garg AV1，Balsitis S1，Durantel D2，Zoulim F3，Delaney WE 4th1，Fletcher SP4。
作者信息

1
    吉列德科学公司，美国加州福斯特市。
2
    INSERM 1052，里昂大学Claude Bernard大学1，CNRS 5286，LéonBérard中心，里昂Cancérologie中心研究中心，法国里昂69003。
3
    INSERM 1052，里昂大学克劳德·伯纳德大学1，法国国家科学研究中心5286，莱昂·贝拉德中心，法国里昂能源研究中心，法国里昂69003;法国里昂的公民医院（HCL），69002法国里昂;法国大学法学院（IUF），75005法国巴黎。
4
    吉列德科学公司，美国加州福斯特市。电子地址：[email protected]。

抽象
背景与目的：

Toll样受体7（TLR7）的口服激动剂GS-9620正在临床开发用于治疗慢性乙型肝炎（CHB）。先前显示GS-9620在CHB的土拨鼠和黑猩猩模型中诱导血清病毒DNA和抗原的长时间抑制。在这里我们调查的分子机制，有助于对GS-9620使用体外模型的乙型肝炎病毒（HBV）感染的抗病毒反应。
方法：

用HBV感染冷冻保存的原代人肝细胞（PHH）和分化的HepaRG（dHepaRG）细胞，并用GS-9620（来自GS-9620处理的人外周血单核细胞（PBMC）的条件培养基（GS-9620条件培养基; GS- 9620-CM）或其他先天性免疫刺激。确定了这些药物的抗病毒和转录反应。
结果：

GS-9620在HBV感染的PHH中没有抗病毒活性，与人肝细胞中低水平的TLR7 mRNA表达一致。相比之下，GS-9620-CM通过I型干扰素（IFN）依赖性机制诱导PHH和dHepaRG细胞中HBV DNA，RNA和抗原水平的持续降低。 GS-9620-CM不降低任一细胞类型的cccDNA水平。转录谱分析表明，GS-9620-CM强烈诱导多种HBV限制性因子 - 尽管不是APOBEC3A或Smc5 / 6复合体 - 并且表明建立的HBV感染不调节冷冻保存的PHH中的先天免疫感应或信号。 GS-9620-CM还诱导了免疫蛋白酶体亚基的表达，并增强了HBV感染PHH中免疫显性病毒肽的表达。
结论：

由GS-9620诱导的I型IFN在人肝细胞中持续抑制HBV而不降低cccDNA水平。此外，HBV抗原呈递增强，表明TLR7诱导的免疫应答的另外组分在CHB动物模型中对GS-9620的抗病毒反应中起作用。
总结：

GS-9620是临床试验用于治疗慢性乙型肝炎病毒（HBV）感染的药物。先前已经证明GS-9620在各种动物模型中抑制HBV，但是潜在的抗病毒机制尚未完全了解。在这项研究中，我们确定GS-9620不直接激活人肝细胞中的抗病毒途径，但是可以通过诱导称为干扰素的抗病毒细胞因子诱导HBV的长时间抑制。然而，干扰素不破坏HBV基因组，表明免疫应答的其他部分（例如激活免疫细胞杀死感染的细胞）也在GS-9620的抗病毒反应中起重要作用。

版权所有©2017欧洲肝脏研究协会。由Elsevier B.V.出版。保留所有权利。
关键词：

APOBEC; GS-9620;乙型肝炎病毒;免疫蛋白酶;干扰素-α;干扰素刺激的基因; MHC; SMC5 / 6; TLR7; cccDNA的

结论：
    29247725
DOI：
    10.1016 / j.jhep.2017.12.007