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The pro-oncogenic effect of the lncRNA H19 in the development of chronic inflammation-mediated hepatocellular carcinoma

Oncogene volume 40, pages 127–139 (2021)Cite this article

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Abstract

The oncofetal long noncoding RNA (lncRNA) H19 is postnatally repressed in most tissues, and re-expressed in many cancers, including hepatocellular carcinoma (HCC). The role of H19 in carcinogenesis is a subject of controversy. We aimed to examine the role of H19 in chronic inflammation-mediated hepatocarcinogenesis using the Mdr2/Abcb4 knockout (Mdr2-KO) mouse, a well-established HCC model. For this goal, we have generated Mdr2-KO/H19-KO double knockout (dKO) mice and followed spontaneous tumor development in the dKO and control Mdr2-KO mice. Cellular localization of H19 and effects of H19 loss in the liver were determined in young and old Mdr2-KO mice. Tumor incidence and tumor load were both significantly decreased in the liver of dKO versus Mdr2-KO females. The expression levels of H19 and Igf2 were variable in nontumor liver tissues of Mdr2-KO females and were significantly downregulated in most matched tumors. In nontumor liver tissue of aged Mdr2-KO females, H19 was expressed mainly in hepatocytes, and hepatocyte proliferation was increased compared to dKO females. At an early age, dKO females displayed lower levels of liver injury and B-cell infiltration, with higher percentage of binuclear hepatocytes. In human samples, H19 expression was higher in females, positively correlated with cirrhosis (in nontumor liver samples) and negatively correlated with CTNNB1 (beta-catenin) mutations and patients’ survival (in tumors). Our data demonstrate that the lncRNA H19 is pro-oncogenic during the development of chronic inflammation-mediated HCC in the Mdr2-KO mouse model, mainly by increasing liver injury and decreasing hepatocyte polyploidy in young mice.

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Fig. 1: H19 loss impeded hepatocarcinogenesis in Mdr2-KO females.
Fig. 2: In the liver of old Mdr2-KO females, H19 is expressed mainly in hepatocytes.
Fig. 3: H19 loss reduces liver injury and B-cell infiltration into the liver of 3-month-old Mdr2-KO females.
Fig. 4: H19 loss increases proportion of binuclear hepatocytes in the liver of 3-month-old Mdr2-KO females.
Fig. 5: Transcriptome profiling of livers from young and old dKO versus Mdr2-KO females using RNA sequencing and GSEA reveals decreased inflammation and liver injury in dKO mice.
Fig. 6: H19 expression in adjacent nontumor liver of HCC patients.
Fig. 7: H19 expression in human HCC tumors.
Fig. 8: In the Mdr2-KO model of chronic inflammation-mediated HCC, H19 has a pro-oncogenic role.

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Acknowledgements

The authors thank Sharona Elgavish, Yuval Nevo, and Hadar Benyamini (Bioinformatics Unit of the I-CORE Computation Center at The Hebrew University and Hadassah, Jerusalem, Israel) for single-cell transcriptomic (SE & YN) and gene set enrichment (HB) analyses, Zakharia Manevich (The Core Research Facility at The Faculty of Medicine, Ein Kerem, The Hebrew University, Jerusalem, Israel) for advice with confocal microscopy, Luisa Dandolo (Institut Cochin, Paris, France) for providing the 129Sv H19Δ3 mutant mice, Shalev Itzkovitz (Weizmann Institute of Science, Rehovot, Israel) for useful discussions and help with smRNA-FISH, Ilan Stein and Yoganathan Krishnamoorthy (Department of Pathology, Hebrew University-Hadassah Medical School) for advice with immunohistochemistry, and Hilla Giladi (The Goldyne Savad Institute of Gene Therapy) for critical reading of the manuscript.

Funding

Robert H. Benson Living Trust and Selma Kron Foundation to student fellowships. DSG is supported by the Kamea Scientific Foundation of the Israeli Government. JHA was supported by ISF 923/14. SC was supported by a funding from Labex OncoImunology and CARPEM. The JZR group was supported by INSERM, Ligue Nationale contre le Cancer (Equipe Labellisée), Labex OncoImmunology (investissement d’avenir), grant IREB, Coup d’Elan de la Fondation Bettencourt-Schueller, the SIRIC CARPEM, Raymond Rosen Award from the Fondation pour le Recherche Médicale, Prix René and Andrée Duquesne—Comité de Paris Ligue Contre le Cancer and Fondation Mérieux. The work of EG was supported by the: ERC advance—GA No. 786575—RxmiRcanceR, Deutsche Forschungsgemeinschaft (DFG) SFB841 project C3, NIH CA197081-02, MOST, ISF collaboration with Canada (2473/2017), personal ISF (486/2017), ICORE–ISF (41/2011), and by DKFZ-MOST.

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Authors and Affiliations

  1. The Goldyne Savad Institute of Gene and Cell Therapy, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    Lika Gamaev, Lina Mizrahi, Tomer Friehmann, Nofar Rosenberg, Devorah Olam, Evelyne Zeira, Jonathan H. Axelrod, Eithan Galun & Daniel S. Goldenberg

  2. Department of Pathology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    Orit Pappo

  3. Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel

    Keren Bahar Halpern,

  4. Centre de Recherche des Cordeliers, INSERM, Sorbonne Université, Université de Paris, Functional Genomics of Solid Tumors Laboratory, F-75006, Paris, France

    Stefano Caruso & Jessica Zucman-Rossi

  5. Hôpital Européen Georges Pompidou, AP-HP, F-75015, Paris, France

    Jessica Zucman-Rossi

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  1. Lika Gamaev
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Contributions

LG and LM: main part of the experimental work and discussions; TF and EZ: data acquisition; NR: data acquisition (single-cell analysis); OP: data analysis (pathology); DO: assistance with mice and liver samples processing; KBH: help with smRNA-FISH and discussions; SC and JZ-R: human HCC data analysis and discussions; JHA: help with mice, discussions, manuscript editing; EG: study idea and discussions, manuscript editing; DSG: study design, data acquisition and analysis, manuscript writing, and discussions.

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Correspondence to Daniel S. Goldenberg.

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Gamaev, L., Mizrahi, L., Friehmann, T. et al. The pro-oncogenic effect of the lncRNA H19 in the development of chronic inflammation-mediated hepatocellular carcinoma. Oncogene 40, 127–139 (2021). https://doi.org/10.1038/s41388-020-01513-7

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