Increased liver stiffness promotes hepatitis B progression by impairing innate immunity in CCl4-induced fibrotic HBV+ transgenic mice
BackgroundHepatitis B virus (HBV) infection develops as an acute or chronic liver disease, which progresses from steatosis, hepatitis, and fibrosis to end-stage liver diseases such as cirrhosis and hepatocellular carcinoma (HCC). An increased stromal stiffness accompanies fibrosis in chronic liver d...
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| Format: | Article |
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Frontiers Media S.A.
2023-08-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1166171/full |
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| author | Grace Bybee Grace Bybee Youra Moeun Weimin Wang Kusum K. Kharbanda Kusum K. Kharbanda Larisa Y. Poluektova Srivatsan Kidambi Natalia A. Osna Natalia A. Osna Murali Ganesan Murali Ganesan |
| author_facet | Grace Bybee Grace Bybee Youra Moeun Weimin Wang Kusum K. Kharbanda Kusum K. Kharbanda Larisa Y. Poluektova Srivatsan Kidambi Natalia A. Osna Natalia A. Osna Murali Ganesan Murali Ganesan |
| author_sort | Grace Bybee |
| collection | DOAJ |
| description | BackgroundHepatitis B virus (HBV) infection develops as an acute or chronic liver disease, which progresses from steatosis, hepatitis, and fibrosis to end-stage liver diseases such as cirrhosis and hepatocellular carcinoma (HCC). An increased stromal stiffness accompanies fibrosis in chronic liver diseases and is considered a strong predictor for disease progression. The goal of this study was to establish the mechanisms by which enhanced liver stiffness regulates HBV infectivity in the fibrotic liver tissue.MethodsFor in vitro studies, HBV-transfected HepG2.2.15 cells were cultured on polydimethylsiloxane gels coated by polyelectrolyte multilayer films of 2 kPa (soft) or 24 kPa (stiff) rigidity mimicking the stiffness of the healthy or fibrotic liver. For in vivo studies, hepatic fibrosis was induced in C57Bl/6 parental and HBV+ transgenic (HBVTg) mice by injecting CCl4 twice a week for 6 weeks.ResultsWe found higher levels of HBV markers in stiff gel-attached hepatocytes accompanied by up-regulated OPN content in cell supernatants as well as suppression of anti-viral interferon-stimulated genes (ISGs). This indicates that pre-requisite “fibrotic” stiffness increases osteopontin (OPN) content and releases and suppresses anti-viral innate immunity, causing a subsequent rise in HBV markers expression in hepatocytes. In vitro results were corroborated by data from HBVTg mice administered CCl4 (HBVTg CCl4). These mice showed higher HBV RNA, DNA, HBV core antigen (HBcAg), and HBV surface antigen (HBsAg) levels after liver fibrosis induction as judged by a rise in Col1a1, SMA, MMPs, and TIMPs mRNAs and by increased liver stiffness. Importantly, CCl4-induced the pro-fibrotic activation of liver cells, and liver stiffness was higher in HBVTg mice compared with control mice. Elevation of HBV markers and OPN levels corresponded to decreased ISG activation in HBVTg CCl4 mice vs HBVTg control mice.ConclusionBased on our data, we conclude that liver stiffness enhances OPN levels to limit anti-viral ISG activation in hepatocytes and promote an increase in HBV infectivity, thereby contributing to end-stage liver disease progression. |
| first_indexed | 2024-03-12T17:43:45Z |
| format | Article |
| id | doaj.art-7dce5104bbc04432b98de9d85196fe16 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-12T17:43:45Z |
| publishDate | 2023-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-7dce5104bbc04432b98de9d85196fe162023-08-03T19:22:48ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-08-011410.3389/fimmu.2023.11661711166171Increased liver stiffness promotes hepatitis B progression by impairing innate immunity in CCl4-induced fibrotic HBV+ transgenic miceGrace Bybee0Grace Bybee1Youra Moeun2Weimin Wang3Kusum K. Kharbanda4Kusum K. Kharbanda5Larisa Y. Poluektova6Srivatsan Kidambi7Natalia A. Osna8Natalia A. Osna9Murali Ganesan10Murali Ganesan11Research Service, Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE, United StatesDepartment of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, United StatesDepartment of Chemical and Biomolecular Engineering, University of Nebraska at Lincoln, Lincoln, NE, United StatesDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, United StatesResearch Service, Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE, United StatesDepartment of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, United StatesDepartment of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, United StatesDepartment of Chemical and Biomolecular Engineering, University of Nebraska at Lincoln, Lincoln, NE, United StatesResearch Service, Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE, United StatesDepartment of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, United StatesResearch Service, Veterans Affairs Nebraska-Western Iowa Health Care System, Omaha, NE, United StatesDepartment of Internal Medicine, University of Nebraska Medical Center, Omaha, NE, United StatesBackgroundHepatitis B virus (HBV) infection develops as an acute or chronic liver disease, which progresses from steatosis, hepatitis, and fibrosis to end-stage liver diseases such as cirrhosis and hepatocellular carcinoma (HCC). An increased stromal stiffness accompanies fibrosis in chronic liver diseases and is considered a strong predictor for disease progression. The goal of this study was to establish the mechanisms by which enhanced liver stiffness regulates HBV infectivity in the fibrotic liver tissue.MethodsFor in vitro studies, HBV-transfected HepG2.2.15 cells were cultured on polydimethylsiloxane gels coated by polyelectrolyte multilayer films of 2 kPa (soft) or 24 kPa (stiff) rigidity mimicking the stiffness of the healthy or fibrotic liver. For in vivo studies, hepatic fibrosis was induced in C57Bl/6 parental and HBV+ transgenic (HBVTg) mice by injecting CCl4 twice a week for 6 weeks.ResultsWe found higher levels of HBV markers in stiff gel-attached hepatocytes accompanied by up-regulated OPN content in cell supernatants as well as suppression of anti-viral interferon-stimulated genes (ISGs). This indicates that pre-requisite “fibrotic” stiffness increases osteopontin (OPN) content and releases and suppresses anti-viral innate immunity, causing a subsequent rise in HBV markers expression in hepatocytes. In vitro results were corroborated by data from HBVTg mice administered CCl4 (HBVTg CCl4). These mice showed higher HBV RNA, DNA, HBV core antigen (HBcAg), and HBV surface antigen (HBsAg) levels after liver fibrosis induction as judged by a rise in Col1a1, SMA, MMPs, and TIMPs mRNAs and by increased liver stiffness. Importantly, CCl4-induced the pro-fibrotic activation of liver cells, and liver stiffness was higher in HBVTg mice compared with control mice. Elevation of HBV markers and OPN levels corresponded to decreased ISG activation in HBVTg CCl4 mice vs HBVTg control mice.ConclusionBased on our data, we conclude that liver stiffness enhances OPN levels to limit anti-viral ISG activation in hepatocytes and promote an increase in HBV infectivity, thereby contributing to end-stage liver disease progression.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1166171/fullHBVliver stiffnessinnate immunityalcoholosteopontin (OPN) |
| spellingShingle | Grace Bybee Grace Bybee Youra Moeun Weimin Wang Kusum K. Kharbanda Kusum K. Kharbanda Larisa Y. Poluektova Srivatsan Kidambi Natalia A. Osna Natalia A. Osna Murali Ganesan Murali Ganesan Increased liver stiffness promotes hepatitis B progression by impairing innate immunity in CCl4-induced fibrotic HBV+ transgenic mice Frontiers in Immunology HBV liver stiffness innate immunity alcohol osteopontin (OPN) |
| title | Increased liver stiffness promotes hepatitis B progression by impairing innate immunity in CCl4-induced fibrotic HBV+ transgenic mice |
| title_full | Increased liver stiffness promotes hepatitis B progression by impairing innate immunity in CCl4-induced fibrotic HBV+ transgenic mice |
| title_fullStr | Increased liver stiffness promotes hepatitis B progression by impairing innate immunity in CCl4-induced fibrotic HBV+ transgenic mice |
| title_full_unstemmed | Increased liver stiffness promotes hepatitis B progression by impairing innate immunity in CCl4-induced fibrotic HBV+ transgenic mice |
| title_short | Increased liver stiffness promotes hepatitis B progression by impairing innate immunity in CCl4-induced fibrotic HBV+ transgenic mice |
| title_sort | increased liver stiffness promotes hepatitis b progression by impairing innate immunity in ccl4 induced fibrotic hbv transgenic mice |
| topic | HBV liver stiffness innate immunity alcohol osteopontin (OPN) |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1166171/full |
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