Activation of Stimulator of Interferon Genes in Hepatocytes Suppresses the Replication of Hepatitis B Virus

Guo, Fang; Tang, Liudi; Shu, Sainan; Sehgal, Mohit; Sheraz, Muhammad; Liu, Bowei; Zhao, Qiong; Cheng, Junjun; Zhao, Xiongyan; Zhou, Tianlun; Chang, Jinhong; Guo, Ju‐Tao

doi:10.1128/aac.00771-17

Cited by 58 publications

(76 citation statements)

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“…In order to establish a highly sensitive cell-based assay for high throughput screening of small molecular compounds that activate human STING, we first established a HepAD38-derived cell line that constitutively expresses human cGAS and STING, which successfully reconstituted a functional cytoplasmic DNA sensor pathway (Guo et al, 2017). Next, we introduced human ISG54 promoter-driven firefly luciferase expression cassette into the HepAD38/cGAS-STING cell line via lentiviral vector transduction and obtained the cell line HepAD38/cGAS-STING/ISG54Luc.…”

Section: Resultsmentioning

confidence: 99%

“…HepAD38 is a HepG2-derived stable cell line supporting a tetracycline (tet)-inducible replication of hepatitis B virus (HBV) and was maintained in DMEM/F12 medium supplemented with 10% fetal bovine serum, 400 μg/ml of G418 and 1 μg/ml tetracycline (Ladner et al, 1997). HepAD38/cGAS-STING and HepAD38/cGAS-STINGΔC are HepAD38-derived cell line constitutively expressing human cGAS and STING or a mutant STING with deletion of 39 amino acid residues from carboxyl terminus (STINGΔC) and were maintained in DMEM/F12 medium supplemented with 10% fetal bovine serum, 400 μg/ml of G418, 2 μg/ml of puromycin and 1 μg/ml tetracycline (Guo et al, 2017). Vero (green monkey kidney) cells were maintained in DMEM (Corning) supplemented with 10% fetal bovine serum.…”

Section: Methodsmentioning

confidence: 99%

“…G10, a small molecule human STING agonist (Sali et al, 2015), served as a control compound and was purchased from Aobious. Double strand DNA 90 (dsDNA-90) was prepared as previously described (Guo et al, 2017). Dispiro diketopiperzine (DSDP) was purchased from Maybridge.…”

Section: Methodsmentioning

confidence: 99%

“…HepAD38/cGAS-STING cell line was originally established for investigation of HBV interaction with cGAS-STING DNA sensor pathway (Guo et al, 2017). However, for screening of cGAS and or STING agonists, the HepAD38/cGAS-STING/ISG54 reporter cell line was cultured in complete DMEM/F12 medium containing 1μg/ml tet throughout the experimental period, which completely prevents HBV replication (Ladner et al, 1997).…”

Section: Methodsmentioning

confidence: 99%

“…IFN-β mRNA, TNFα mRNA, IL-28A, IL-29 mRNA and IL-6 mRNA were measured by quantitative one-step RT-PCR (qRT-PCR) assays using primers reported previously (Guo et al, 2017). β-actin mRNA served as internal control to normalize cytokine mRNAs.…”

Section: Methodsmentioning

confidence: 99%

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A cell-based high throughput screening assay for the discovery of cGAS-STING pathway agonists

et al. 2017

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Stimulator of interferon genes (STING) is an endoplasmic reticulum transmembrane protein that serves as a molecular hub for activation of interferon and inflammatory cytokine response by multiple cellular DNA sensors. Not surprisingly, STING has been demonstrated to play an important role in host defense against microorganisms and pharmacologic activation of STING is considered as an attractive strategy to treat viral diseases and boost antitumor immunity. In light of this we established a HepAD38-derived reporter cell line that expresses firefly luciferase in response to the activation of cyclic GMP-AMP synthase (cGAS)-STING pathway for high throughput screening (HTS) of small molecular human STING agonists. This cell-based reporter assay required only 4 h treatment with a reference STING agonist to induce a robust luciferase signal and was demonstrated to have an excellent performance in HTS format. By screening 16,000 compounds, a dispiro diketopiperzine (DSDP) compound was identified to induce proinflammatory cytokine response in a manner dependent on the expression of functional human STING, but not mouse STING. Moreover, we showed that DSDP induced an interferon-dominant cytokine response in human skin fibroblasts and peripheral blood mononuclear cells, which in turn potently suppressed the replication of yellow fever virus, dengue virus and Zika virus. We have thus established a robust cell-based assay system suitable for rapid discovery and mechanistic analyses of cGAS-STING pathway agonists. Identification of DSDP as a human STING agonist enriches the pipelines of STING-targeting drug development for treatment of viral infections and cancers.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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A cell-based high throughput screening assay for the discovery of cGAS-STING pathway agonists

et al. 2017

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Cross talk between hepatitis B virus and innate immunity of hepatocytes

Golsaz‐Shirazi

Shokri

2021

Reviews in Medical Virology

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Innate immunity plays a major role in controlling viral infections. Recent exploration of sodium taurocholate co-transporting polypeptide receptor as specific hepatitis B virus (HBV) receptor in human hepatocytes has provided appropriate cell culture tools to study the innate immunity of hepatocytes and its cross talk with HBV. In this review, we give a brief update on interaction between HBV and innate immunity using the currently available in vitro cellular models that support the complete life cycle of HBV. We will discuss how HBV can act as a 'stealth' virus to counteract the innate immune responses mediated by the pathogen recognition receptors of hepatocytes and escape the first line of surveillance of the host immune system. We give an overview of the cellular components of innate immunity that present in these in vitro models and discuss how activating these innate immunity components may contribute to the eradication of HBV infection. K E Y W O R D Scell culture models, hepatitis B virus (HBV), innate immunity, pattern recognition receptors, toll-like receptors | INTRODUCTIONHepatitis B virus (HBV) selectively infects hepatocytes of human liver. 1,2 Despite the effectiveness of hepatitis B vaccination, HBV is still a major public health problem with more than 240 million chronic carriers worldwide of whom almost a third are at high risk of developing serious HBV-related complications, such as liver cirrhosis and hepatocellular carcinoma (HCC). 3,4 The first line of defence against viruses is innate immunity. Most viruses are sensed and detected by innate immunity at early stages of infection, leading to initiation of anti-viral responses such as production of interferons (IFNs) and other pro-inflammatory cytokines. [5][6][7] Induction of these cytokines during viral infection occurs through two distinct signalling pathways: subfamilies of toll-like receptors (TLRs) as well as other innate immunity signalling pathways that sense RNA or DNA of viruses, such as retinoic acid inducible gene I (RIG-I)/melanoma differentiation-associated protein 5 (MDA5) and cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS)/STING. 6,8 However, HBV has been considered a 'stealth' virus and before hepatocytes can sense and respond to the

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STING and liver disease

Chen

Yang

2021

J Gastroenterol

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STING (stimulator of interferon genes) also known as transmembrane protein 173 (TMEM173) is a cytoplasmic DNA sensor which can be activated by the upstream cyclic dinucleotides (CDNs). This activation produces cytokines such as interferons and pro-inflammatory factors via the downstream IRF3 and NF-κB pathways, triggering an innate immune response and adaptive immunity to maintain homeostasis. STING is mainly expressed and activated in non-parenchymal cells, thus exerting a corresponding effect to maintain the homeostasis of the liver. In viral hepatitis, interferons and pro-inflammatory factors produced after STING activation initiate the immune response to inhibit virus replication and assembly. In the case of metabolic diseases of the liver, the activation of STING in kupffer cells and hepatic stellate cells leads to inflammation, the proliferation of connective tissue, and metabolic disorders in the hepatocytes, promoting the occurrence and development of the disease. In hepatocellular carcinoma, STING has two contradictory roles. When STING is activated in dendritic cells and macrophages, a large number of cytokines can be produced to initiate innate immune effects directly and to exert adaptive immunity through the recruitment and activation of T cells; however, aberrant activation of the STING pathway leads to a weakening of immune function and promotes oncogenesis and metastasis. Here, we summarize the interactions between STING and liver disease that have currently been identified and how to achieve therapeutic goals by modulating the activity of the STING pathway.

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Activation of Stimulator of Interferon Genes in Hepatocytes Suppresses the Replication of Hepatitis B Virus

Cited by 58 publications

References 56 publications

A cell-based high throughput screening assay for the discovery of cGAS-STING pathway agonists

A cell-based high throughput screening assay for the discovery of cGAS-STING pathway agonists

Cross talk between hepatitis B virus and innate immunity of hepatocytes

STING and liver disease

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