ATOH8 promotes HBV immune tolerance by inhibiting the pyroptotic pathway in hepatocytes

Liu,Xiaofei; Liu,Xiaofei; Fan,Zhenyu; Fan,Zhenyu; Chen,Liping; Chen,Liping; Yang,Jingmao; Yang,Jingmao; Cheng,Jilin; Cheng,Jilin

doi:10.3892/mmr.2023.13018

ATOH8 promotes HBV immune tolerance by inhibiting the pyroptotic pathway in hepatocytes

Authors:
- Xiaofei Liu
- Zhenyu Fan
- Liping Chen
- Jingmao Yang
- Jilin Cheng
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Affiliations: Department of Gastroenterology and Hepatology, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, P.R. China, Department of Gastroenterology and Hepatology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, P.R. China
Published online on: May 22, 2023 https://doi.org/10.3892/mmr.2023.13018
Article Number: 131
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The mechanism of hepatitis B virus (HBV) immune tolerance remains unclear. Our previous studies showed that ATOH8 plays an important role in the liver tumor immune microenvironment; however, the specific immune regulatory mechanism requires further studies. Studies have shown that the hepatitis C virus (HCV) can cause hepatocyte pyroptosis; however, the relationship between HBV and pyroptosis is contested. Therefore, this study aimed to determine whether ATOH8 interfered with HBV activity through pyroptosis to further study the mechanism of ATOH8 on immune regulation and enrich our understanding of HBV‑induced invasion. The expression levels of pyroptosis‑related molecules (GSDMD and Caspase‑1) in liver cancer tissues and peripheral blood mononuclear cells (PBMCs) of patients with HBV were assessed using qPCR and western blotting. HepG2.2.15 and Huh7 cells were used to overexpress ATOH8 using a recombinant lentiviral vector. The HBV DNA expression levels in HepG2.2.15 cells were detected using absolute quantitative (q)PCR, and the hepatitis B surface antigen expression levels in the HepG2.2.15 cell culture supernatant were measured using ELISA. The expression of pyroptosis‑related molecules in Huh7 and HepG2.2.15 cells was detected using western blotting and qPCR. Additionally, the expression levels of inflammatory factors including TNF‑α, INF‑α, IL‑18, and IL‑1β were detected using qPCR and ELISA. The liver cancer tissues and PBMCs of patients with HBV showed higher expressions of pyroptosis‑related molecules than those of normal samples. ATOH8‑overexpressed HepG2.2.15 cells had higher HBV expression levels but lower levels of pyroptosis‑related molecules, such as GSDMD and Caspase‑1, than those in the control group. Similarly, the expression levels of pyroptosis‑related molecules in Huh7 cells overexpressing ATOH8 were lower than that in Huh7‑GFP cells. Further detection of the expression of INF‑α and TNF‑α in HepG2.2.15 cells overexpressing ATOH8 showed that ATOH8 overexpression increased the expression of these inflammatory factors, including those associated with pyroptosis (IL‑18 and IL‑1β). In conclusion, ATOH8 promoted HBV immune escape by inhibiting hepatocyte pyroptosis.

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1	Nicolini LA, Orsi A, Tatarelli P, Viscoli C, Icardi G and Sticchi L: A global view to HBV chronic infection: Evolving strategies for diagnosis, treatment and prevention in immunocompetent individuals. Int J Environ Res Public Health. 16:33072019. View Article : Google Scholar : PubMed/NCBI
2	Pollicino T and Caminiti G: HBV–Integration studies in the clinic: Role in the natural history of infection. Viruses. 13:3682021. View Article : Google Scholar : PubMed/NCBI
3	Zhou L, He R, Fang P, Li M, Yu H, Wang Q, Yu Y, Wang F, Zhang Y, Chen A, et al: Hepatitis B virus rigs the cellular metabolome to avoid innate immune recognition. Nat Commun. 12:982021. View Article : Google Scholar : PubMed/NCBI
4	Wieland S, Thimme R, Purcell RH and Chisari FV: Genomic analysis of the host response to hepatitis B virus infection. Proc Natl Acad Sci USA. 101:6669–6674. 2004. View Article : Google Scholar : PubMed/NCBI
5	Lebossé F, Testoni B, Fresquet J, Facchetti F, Galmozzi E, Fournier M, Hervieu V, Berthillon P, Berby F, Bordes I, et al: Intrahepatic innate immune response pathways are downregulated in untreated chronic hepatitis B. J Hepatol. 66:897–909. 2017. View Article : Google Scholar : PubMed/NCBI
6	Luangsay S, Gruffaz M, Isorce N, Testoni B, Michelet M, Faure-Dupuy S, Maadadi S, Ait-Goughoulte M, Parent R, Rivoire M, et al: Early inhibition of hepatocyte innate responses by hepatitis B virus. J Hepatol. 63:1314–1322. 2015. View Article : Google Scholar : PubMed/NCBI
7	Visvanathan K, Skinner NA, Thompson AJV, Riordan SM, Sozzi V, Edwards R, Rodgers S, Kurtovic J, Chang J, Lewin S, et al: Regulation of toll-like receptor-2 expression in chronic hepatitis B by the precore protein. Hepatology. 45:102–110. 2007. View Article : Google Scholar : PubMed/NCBI
8	Li M, Sun R, Xu L, Yin W, Chen Y, Zheng X, Lian Z, Wei H and Tian Z: Kupffer cells support hepatitis B virus-mediated CD8+ T cell exhaustion via hepatitis B core antigen-TLR2 interactions in mice. J Immunol. 195:3100–3109. 2015. View Article : Google Scholar : PubMed/NCBI
9	Teng Y, Xu Z, Zhao K, Zhong Y, Wang J, Zhao L, Zheng Z, Hou W, Zhu C, Chen X, et al: Novel function of SART1 in HNF4α transcriptional regulation contributes to its antiviral role during HBV infection. J Hepatol. 75:1072–1082. 2021. View Article : Google Scholar : PubMed/NCBI
10	Zhang Y, Chen X, Cao Y and Yang Z: Roles of APOBEC3 in hepatitis B virus (HBV) infection and hepatocarcinogenesis. Bioengineered. 12:2074–2086. 2021. View Article : Google Scholar : PubMed/NCBI
11	Lak R, Yaghobi R and Garshasbi M: Importance of miR-125a-5p and miR-122-5p expression in patients with HBV infection. Cell Mol Biol (Noisy-le-grand). 66:1–8. 2020. View Article : Google Scholar : PubMed/NCBI
12	Oropeza CE, Tarnow G, Sridhar A, Taha TY, Shalaby RE and McLachlan A: The regulation of HBV transcription and replication. Adv Exp Med Biol. 1179:39–69. 2020. View Article : Google Scholar : PubMed/NCBI
13	Xu M, Huang S, Dong X, Chen Y, Li M, Shi W, Wang G, Huang C, Wang Q, Liu Y, et al: A novel isoform of ATOH8 promotes the metastasis of breast cancer by regulating RhoC. J Mol Cell Biol. 13:59–71. 2021. View Article : Google Scholar : PubMed/NCBI
14	Song Y, Pan G, Chen L, Ma S, Zeng T, Chan THM, Li L, Lian Q, Chow R, Cai X, et al: Loss of ATOH8 increases stem cell features of hepatocellular carcinoma cells. Gastroenterology. 149:1068–1081.e5. 2015. View Article : Google Scholar : PubMed/NCBI
15	Chen L, Yang J, Wang Y, Wu N, Li X, Li J, Huang Y and Cheng J: ATOH8 overexpression inhibits the tumor progression and monocyte chemotaxis in hepatocellular carcinoma. Int J Clin Exp Pathol. 13:2534–2543. 2020.PubMed/NCBI
16	Yu P, Zhang X, Liu N, Tang L, Peng C and Chen X: Pyroptosis: Mechanisms and diseases. Signal Transduct Target Ther. 6:1282021. View Article : Google Scholar : PubMed/NCBI
17	Song L, Pei L, Yao S, Wu Y and Shang Y: NLRP3 inflammasome in neurological diseases, from functions to therapies. Front Cell Neurosci. 11:632017. View Article : Google Scholar : PubMed/NCBI
18	Pezuk JA: Pyroptosis in combinatorial treatment to improve cancer patients' outcome, is that what we want? EBioMedicine. 41:17–18. 2019. View Article : Google Scholar : PubMed/NCBI
19	Zhang L, Zeyu W, Liu B, Jang S, Zhang Z and Jiang Y: Pyroptosis in liver disease. Rev Esp Enferm Dig. 113:280–285. 2021.PubMed/NCBI
20	Ługowska A, Hetmańczyk-Sawicka K, Iwanicka-Nowicka R, Fogtman A, Cieśla J, Purzycka-Olewiecka JK, Sitarska D, Płoski R, Filocamo M, Lualdi S, et al: Gene expression profile in patients with Gaucher disease indicates activation of inflammatory processes. Sci Rep. 9:60602019. View Article : Google Scholar : PubMed/NCBI
21	Jiang Y, Han Q, Zhao H and Zhang J: The mechanisms of HBV-induced hepatocellular carcinoma. J Hepatocell Carcinoma. 8:435–450. 2021. View Article : Google Scholar : PubMed/NCBI
22	Yu X, Lan P, Hou X, Han Q, Lu N, Li T, Jiao C, Zhang J, Zhang C and Tian Z: HBV inhibits LPS-induced NLRP3 inflammasome activation and IL-1β production via suppressing the NF-κB pathway and ROS production. J Hepatol. 66:693–702. 2017. View Article : Google Scholar : PubMed/NCBI
23	Xie WH, Ding J, Xie XX, Yang XH, Wu XF, Chen ZX, Guo QL, Gao WY, Wang XZ and Li D: Hepatitis B virus X protein promotes liver cell pyroptosis under oxidative stress through NLRP3 inflammasome activation. Inflamm Res. 69:683–696. 2020. View Article : Google Scholar : PubMed/NCBI
24	Wei Q, Mu K, Li T, Zhang Y, Yang Z, Jia X, Zhao W, Huai W, Guo P and Han L: Deregulation of the NLRP3 inflammasome in hepatic parenchymal cells during liver cancer progression. Lab Invest. 94:52–62. 2014. View Article : Google Scholar : PubMed/NCBI
25	Zarković G: Population growth and planning in Yugoslavia. Nar Zdrav. 28:41–48. 1972.(In Serbian). PubMed/NCBI
26	Whitsett TL, Levin DC and Manion CV: Comparison of the beta 1 and beta 2 adrenoceptor blocking properties of acebutolol and propranolol. Chest. 82:668–673. 1982. View Article : Google Scholar : PubMed/NCBI