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Joining the dots – NEDDylation in cancer cells regulates the tumour environment in cholangiocarcinoma

  • Tom Lüdde
    Correspondence
    Corresponding authors. Addresses: Clinic for Gastroenterology, Hepatology and Infectious Diseases, University Hospital Düsseldorf, Medical Faculty of Heinrich Heine, University Düsseldorf, 40225 Düsseldorf, Germany
    Affiliations
    Clinic for Gastroenterology, Hepatology and Infectious Diseases, University Hospital Düsseldorf, Medical Faculty of Heinrich Heine, University Düsseldorf, 40225 Düsseldorf, Germany
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  • Luke Boulter
    Correspondence
    MRC Human Genetics Unit, Institute of Genetics and Cancer, Edinburgh, EH4 2XU, United Kingdom
    Affiliations
    MRC Human Genetics Unit, Institute of Genetics and Cancer, Edinburgh, EH4 2XU, United Kingdom
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Published:April 28, 2022DOI:https://doi.org/10.1016/j.jhep.2022.04.018

      Linked Article

      See Article, pages 177–190
      Solid tumours are a highly complex environment in which a multitude of cell types interact to promote disease progression and evade therapeutic intervention. Whilst much of the work investigating cancer has concentrated on the tumour cells themselves, understanding the role of the tumour microenvironment has gained increasing traction over the last decade and targeting this component of the tumour has been subject to renewed vigour, particularly in those cancers where therapies developed to clear tumour cells have had marginal impact.
      The presence of a complex stroma, the fibrogenic, endothelial and immunological support structure of a tumour, is a hallmark of cholangiocarcinoma (CCA) where non-cancer cells can comprise up to 70% of the tumour mass.
      • Banales J.M.
      • Marin J.J.G.
      • Lamarca A.
      • Rodrigues P.M.
      • Khan S.A.
      • Roberts L.R.
      • et al.
      Cholangiocarcinoma 2020: the next horizon in mechanisms and management.
      The presence of this complex stroma is thought to contribute to the highly aggressive nature of this cancer and contributes to poor survival (∼5% over 5 years) following diagnosis and wide metastatic dispersal in advanced disease.
      • Brindley P.J.
      • Bachini M.
      • Ilyas S.I.
      • Khan S.A.
      • Loukas A.
      • Sirica A.E.
      • et al.
      Cholangiocarcinoma.
      Increasingly, the cellular and non-cellular components of the CCA stroma are being defined, but our understanding of the crosstalk between tumour cells and the fibro-immune microenvironment in CCA is lacking. In both premalignant bile duct diseases (including those that increase the risk of CCA, such as primary sclerosing cholangitis) and in CCA itself, work from a range of laboratories has demonstrated that the biliary epithelium can communicate out to its microenvironment to regulate immune cell recruitment,
      • Guillot A.
      • Guerri L.
      • Feng D.
      • Kim S.-J.
      • Ahmed Y.A.
      • Paloczi J.
      • et al.
      Bile acid-activated macrophages promote biliary epithelial cell proliferation through integrin αvβ6 upregulation following liver injury.
      fibroblast activation
      • Lobe C.
      • Vallette M.
      • Arbelaiz A.
      • Gonzalez-Sanchez E.
      • Izquierdo L.
      • Pellat A.
      • et al.
      Zinc finger E-box binding homeobox 1 promotes cholangiocarcinoma progression through tumor dedifferentiation and tumor-stroma paracrine signaling.
      and directly influence the deposition of a new “proliferative” extracellular matrix by indirectly communicating with non-epithelial cells.
      • Wilson D.H.
      • Jarman E.J.
      • Mellin R.P.
      • Wilson M.L.
      • Waddell S.H.
      • Tsokkou P.
      • et al.
      Non-canonical Wnt signalling regulates scarring in biliary disease via the planar cell polarity receptors.
      Similarly, the formation of the regenerative and cancer niche promotes progenitor traits in both biliary epithelial cells and their cancerous counterparts and likely enables both their proliferation during disease and also promotes their immune evasion.
      • Thongchot S.
      • Vidoni C.
      • Ferraresi A.
      • Loilome W.
      • Khuntikeo N.
      • Sangkhamanon S.
      • et al.
      Cancer-associated fibroblast-derived IL-6 determines unfavorable prognosis in cholangiocarcinoma by affecting autophagy-associated chemoresponse.
      ,
      • Lin Y.
      • Cai Q.
      • Chen Y.
      • Shi T.
      • Liu W.
      • Mao L.
      • et al.
      CAFs shape myeloid-derived suppressor cells to promote stemness of intrahepatic cholangiocarcinoma through 5-lipoxygenase.
      In order to better understand how cancer cells communicate with their tumour stroma, we not only need to understand the signals that are produced by the cancerous epithelium in CCA and define how these communicate with the local environment, but we must also determine what intracellular processes are occurring in the cancer cells themselves to facilitate tumour-stroma crosstalk. This is not an easy question to address, as the cancer proteome is subject to dynamic regulation at both the translational and post-translational levels.
      • Grosso S.
      • Marini A.
      • Gyuraszova K.
      • Voorde J.V.
      • Sfakianos A.
      • Garland G.D.
      • et al.
      The pathogenesis of mesothelioma is driven by a dysregulated translatome.
      ,
      • Schmidt S.
      • Gay D.
      • Uthe F.W.
      • Denk S.
      • Paauwe M.
      • Matthes N.
      • et al.
      A MYC-GCN2-eIF2α negative feedback loop limits protein synthesis to prevent MYC-dependent apoptosis in colorectal cancer.
      In this issue of Journal of Hepatology, Olaizola and colleagues define how NEDDylation – a post-translational modification where NEDD8 (a ubiquitin-like molecule) is conjugated to its target proteins – regulates tumour progression
      • Ohh M.
      • Kim W.Y.
      • Moslehi J.J.
      • Chen Y.
      • Chau V.
      • Read M.A.
      • et al.
      An intact NEDD8 pathway is required for Cullin-dependent ubiquitylation in mammalian cells.
      (Fig. 1). NEDDylation of a protein often serves to alter the target proteins ability to interact with co-factors or to form complexes with other proteins, thereby affecting multi-protein complex assembly and function. In this report, Olaizola et al. found that, in healthy patient liver tissues, expression of the NEDDylation machinery is low, but in both premalignant (BilINs) and malignant tissue, expression of the NEDD8-specific E1 ligase, NAE1, which is essential for the conjugation of NEDD8 to proteins, is increased in human disease. Critically, when examined further, the authors identify a range of NEDD8-conjugated proteins that are associated with multiple oncogenic processes. The reliance of cancer cells on NEDDylation has been described in other, non-solid and solid malignancies and small molecules have been developed to limit the ability of NAE1 to add NEDD8 to its substrates. For example, pevonedistat is currently in a phase I clinical trial
      • Shoji H.
      • Takahari D.
      • Hara H.
      • Nagashima K.
      • Adachi J.
      • Boku N.
      A Phase I study of pevonedistat plus capecitabine plus oxaliplatin in patients with advanced gastric cancer refractory to platinum (NCCH-1811).
      and acts as a competitive AMP mimetic, thereby limiting the ability of NEDD8-AMP intermediates to form, which is a rate limiting step for NEDDlyation of NAE1 substrates. Olaizola and colleagues make use of the addiction that CCA cells have to protein NEDDlyation and demonstrate when treating human CCA cells in vitro or in xenografts in immunocompromised mice that pevonedistat reduces tumour cell proliferation and tumour progression. While a stalwart model in the field, xenografts do have their limitations and further work will be needed to really understand the interactions between CCA tumour cells and their microenvironment. A number of stroma-targeting approaches are being trialled in CCA and combining pevonedistat with either anti-PD-1/anti-PD-L1 immunotherapy
      • Jakubowski C.D.
      • Azad N.S.
      Immune checkpoint inhibitor therapy in biliary tract cancer (cholangiocarcinoma).
      ,
      • Klein O.
      • Kee D.
      • Nagrial A.
      • Markman B.
      • Underhill C.
      • Michael M.
      • et al.
      Evaluation of combination nivolumab and ipilimumab immunotherapy in patients with advanced biliary tract cancers: subgroup analysis of a phase 2 nonrandomized clinical trial.
      or anti-VEGF therapies such as regorafenib
      • Demols A.
      • Borbath I.
      • Van den Eynde M.
      • Houbiers G.
      • Peeters M.
      • Marechal R.
      • et al.
      Regorafenib after failure of gemcitabine and platinum-based chemotherapy for locally advanced/metastatic biliary tumors: REACHIN, a randomized, double-blind, phase II trial.
      ,
      • Cousin S.
      • Cantarel C.
      • Guegan J.-P.
      • Mazard T.
      • Gomez-Roca C.
      • Metges J.-P.
      • et al.
      Regorafenib-avelumab combination in patients with biliary tract cancer (REGOMUNE): a single-arm, open-label, phase II trial.
      in immunocompetent models could enhance uncoupling of cancer cells from their supportive stroma and further drive tumour regression (Fig. 1).
      Figure thumbnail gr1
      Fig. 1Tumour cells are part of a complex organ comprising tumour cells and supporting cells such as cancer-associated fibroblasts, endothelial cells and immune cells.
      To generate this pathological tissue, cancer cells must alter their proteome and secretome to influence the microenvironment. Part of this regulation occurs through NEDDylation, a process by which proteins are post-translationally modified to alter their stability and ability to form biological protein complexes.
      Whilst these findings are exciting, understanding the fundamental mechanism behind pevonedistat-reduced NEDDylation remains a challenge, as systemic therapies would alter these modifications in both cancer cells and those cells in the stroma that rely on NEDD8 function. To overcome this limitation, the authors make use of a sophisticated approach where they use a genetically modified mouse line in which NAE1 can be specifically deleted in tumour cells. When these animals are given CCA the tumours that form are smaller and have an altered secretome, where many families of pro-cancerous signals including Wnt, TNF and TGFβ are reduced, whilst apoptosis and necrosis are increased. By understanding how NEDDylation regulates cancer cell-stromal interactions, Olaizola et al. have opened a window into the complexity of the tumour organ in CCA.
      Over recent years, the development of precision medicine approaches in CCA have expanded the therapeutic toolkit. The reality remains, however, that even with these targeted therapies (including FGFR
      • Subbiah V.
      • Iannotti N.O.
      • Gutierrez M.
      • Smith D.C.
      • Féliz L.
      • Lihou C.F.
      • et al.
      FIGHT-101, a first-in-human study of potent and selective FGFR 1-3 inhibitor pemigatinib in pan-cancer patients with FGF/FGFR alterations and advanced malignancies.
      and IDH1
      • Lapin M.
      • Huang H.J.
      • Chagani S.
      • Javle M.
      • Shroff R.T.
      • Pant S.
      • et al.
      Monitoring of dynamic changes and clonal evolution in circulating tumor DNA from patients with IDH-mutated cholangiocarcinoma treated with isocitrate dehydrogenase inhibitors.
      -inhibitors) the majority of patients do not carry therapeutically actionable mutations and subsequently the majority receive chemotherapy as standard of care. With advances such as the one described here by Olaizola et al., where pevonedistat can be used to disrupt a fundamental biological function on which cancer cells rely, there is the potential for NEDDylation-inhibitors to be a broad-acting therapeutic in CCA regardless of the underlying genetic status or anatomical location. Furthermore, it is clear that there are groups of patients, such as those diagnosed with primary sclerosing cholangitis
      • Rudolph G.
      • Gotthardt D.
      • Kloeters-Plachky P.
      • Rost D.
      • Kulaksiz H.
      • Stiehl A.
      In PSC with dominant bile duct stenosis, IBD is associated with an increase of carcinomas and reduced survival.
      ,
      • Tischendorf J.J.W.
      • Krüger M.
      • Trautwein C.
      • Duckstein N.
      • Schneider A.
      • Manns M.P.
      • et al.
      Cholangioscopic characterization of dominant bile duct stenoses in patients with primary sclerosing cholangitis.
      (or other pre-malignant diseases including Caroli disease
      • Fard-Aghaie M.H.
      • Makridis G.
      • Reese T.
      • Feyerabend B.
      • Wagner K.C.
      • Schnitzbauer A.
      • et al.
      The rate of cholangiocarcinoma in Caroli Disease A German multicenter study.
      ) who live with a substantially increased risk of developing CCA. For these patients, extended long-term clinical monitoring is essential and the identification of prophylactic therapies that could limit the conversion of diseased ducts into cancer would be a substantial addition to their care. As Olaizola et al. demonstrated, the molecular machinery required for NEDDylation is upregulated in the pre-malignant disease state and, as such, pevonedistat has potential in this setting.

      Financial support

      LB is funded by a personal fellowship from Cancer Research UK (C52499/A27948).

      Authors’ contributions

      The authors contributed equally to the production of this manuscript.

      Conflict of interest

      The authors have no conflicts of interest to declare regarding this editorial.
      Please refer to the accompanying ICMJE disclosure forms for further details.

      Supplementary data

      The following are the supplementary data to this article:

      References

        • Banales J.M.
        • Marin J.J.G.
        • Lamarca A.
        • Rodrigues P.M.
        • Khan S.A.
        • Roberts L.R.
        • et al.
        Cholangiocarcinoma 2020: the next horizon in mechanisms and management.
        Nat Rev Gastroenterol Hepatol. 2020; 17: 557-588https://doi.org/10.1038/s41575-020-0310-z
        • Brindley P.J.
        • Bachini M.
        • Ilyas S.I.
        • Khan S.A.
        • Loukas A.
        • Sirica A.E.
        • et al.
        Cholangiocarcinoma.
        Nat Rev Dis Primers. 2021; 7: 65https://doi.org/10.1038/s41572-021-00300-2
        • Guillot A.
        • Guerri L.
        • Feng D.
        • Kim S.-J.
        • Ahmed Y.A.
        • Paloczi J.
        • et al.
        Bile acid-activated macrophages promote biliary epithelial cell proliferation through integrin αvβ6 upregulation following liver injury.
        J Clin Invest. 2021; : 131https://doi.org/10.1172/JCI132305
        • Lobe C.
        • Vallette M.
        • Arbelaiz A.
        • Gonzalez-Sanchez E.
        • Izquierdo L.
        • Pellat A.
        • et al.
        Zinc finger E-box binding homeobox 1 promotes cholangiocarcinoma progression through tumor dedifferentiation and tumor-stroma paracrine signaling.
        Hepatology. 2021; 74: 3194-3212https://doi.org/10.1002/hep.32069
        • Wilson D.H.
        • Jarman E.J.
        • Mellin R.P.
        • Wilson M.L.
        • Waddell S.H.
        • Tsokkou P.
        • et al.
        Non-canonical Wnt signalling regulates scarring in biliary disease via the planar cell polarity receptors.
        Nat Commun. 2020; 11: 445https://doi.org/10.1038/s41467-020-14283-3
        • Thongchot S.
        • Vidoni C.
        • Ferraresi A.
        • Loilome W.
        • Khuntikeo N.
        • Sangkhamanon S.
        • et al.
        Cancer-associated fibroblast-derived IL-6 determines unfavorable prognosis in cholangiocarcinoma by affecting autophagy-associated chemoresponse.
        Cancers (Basel). 2021; 13https://doi.org/10.3390/cancers13092134
        • Lin Y.
        • Cai Q.
        • Chen Y.
        • Shi T.
        • Liu W.
        • Mao L.
        • et al.
        CAFs shape myeloid-derived suppressor cells to promote stemness of intrahepatic cholangiocarcinoma through 5-lipoxygenase.
        Hepatology. 2022; 75: 28-42https://doi.org/10.1002/hep.32099
        • Grosso S.
        • Marini A.
        • Gyuraszova K.
        • Voorde J.V.
        • Sfakianos A.
        • Garland G.D.
        • et al.
        The pathogenesis of mesothelioma is driven by a dysregulated translatome.
        Nat Commun. 2021; 12: 4920https://doi.org/10.1038/s41467-021-25173-7
        • Schmidt S.
        • Gay D.
        • Uthe F.W.
        • Denk S.
        • Paauwe M.
        • Matthes N.
        • et al.
        A MYC-GCN2-eIF2α negative feedback loop limits protein synthesis to prevent MYC-dependent apoptosis in colorectal cancer.
        Nat Cell Biol. 2019; 21: 1413-1424https://doi.org/10.1038/s41556-019-0408-0
        • Ohh M.
        • Kim W.Y.
        • Moslehi J.J.
        • Chen Y.
        • Chau V.
        • Read M.A.
        • et al.
        An intact NEDD8 pathway is required for Cullin-dependent ubiquitylation in mammalian cells.
        EMBO Rep. 2002; 3: 177-182https://doi.org/10.1093/embo-reports/kvf028
        • Shoji H.
        • Takahari D.
        • Hara H.
        • Nagashima K.
        • Adachi J.
        • Boku N.
        A Phase I study of pevonedistat plus capecitabine plus oxaliplatin in patients with advanced gastric cancer refractory to platinum (NCCH-1811).
        Future Sci OA. 2021; 7: FSO721https://doi.org/10.2144/fsoa-2021-0023
        • Jakubowski C.D.
        • Azad N.S.
        Immune checkpoint inhibitor therapy in biliary tract cancer (cholangiocarcinoma).
        Chin Clin Oncol. 2020; 9: 2https://doi.org/10.21037/cco.2019.12.10
        • Klein O.
        • Kee D.
        • Nagrial A.
        • Markman B.
        • Underhill C.
        • Michael M.
        • et al.
        Evaluation of combination nivolumab and ipilimumab immunotherapy in patients with advanced biliary tract cancers: subgroup analysis of a phase 2 nonrandomized clinical trial.
        JAMA Oncol. 2020; 6: 1405-1409https://doi.org/10.1001/jamaoncol.2020.2814
        • Demols A.
        • Borbath I.
        • Van den Eynde M.
        • Houbiers G.
        • Peeters M.
        • Marechal R.
        • et al.
        Regorafenib after failure of gemcitabine and platinum-based chemotherapy for locally advanced/metastatic biliary tumors: REACHIN, a randomized, double-blind, phase II trial.
        Ann Oncol. 2020; 31: 1169-1177https://doi.org/10.1016/j.annonc.2020.05.018
        • Cousin S.
        • Cantarel C.
        • Guegan J.-P.
        • Mazard T.
        • Gomez-Roca C.
        • Metges J.-P.
        • et al.
        Regorafenib-avelumab combination in patients with biliary tract cancer (REGOMUNE): a single-arm, open-label, phase II trial.
        Eur J Cancer. 2022; 162: 161-169https://doi.org/10.1016/j.ejca.2021.11.012
        • Subbiah V.
        • Iannotti N.O.
        • Gutierrez M.
        • Smith D.C.
        • Féliz L.
        • Lihou C.F.
        • et al.
        FIGHT-101, a first-in-human study of potent and selective FGFR 1-3 inhibitor pemigatinib in pan-cancer patients with FGF/FGFR alterations and advanced malignancies.
        Ann Oncol. 2022; https://doi.org/10.1016/j.annonc.2022.02.001
        • Lapin M.
        • Huang H.J.
        • Chagani S.
        • Javle M.
        • Shroff R.T.
        • Pant S.
        • et al.
        Monitoring of dynamic changes and clonal evolution in circulating tumor DNA from patients with IDH-mutated cholangiocarcinoma treated with isocitrate dehydrogenase inhibitors.
        JCO Precis Oncol. 2022; 6e2100197https://doi.org/10.1200/PO.21.00197
        • Rudolph G.
        • Gotthardt D.
        • Kloeters-Plachky P.
        • Rost D.
        • Kulaksiz H.
        • Stiehl A.
        In PSC with dominant bile duct stenosis, IBD is associated with an increase of carcinomas and reduced survival.
        J Hepatol. 2010; 53: 313-317https://doi.org/10.1016/j.jhep.2010.02.030
        • Tischendorf J.J.W.
        • Krüger M.
        • Trautwein C.
        • Duckstein N.
        • Schneider A.
        • Manns M.P.
        • et al.
        Cholangioscopic characterization of dominant bile duct stenoses in patients with primary sclerosing cholangitis.
        Endoscopy. 2006; 38: 665-669https://doi.org/10.1055/s-2006-925257
        • Fard-Aghaie M.H.
        • Makridis G.
        • Reese T.
        • Feyerabend B.
        • Wagner K.C.
        • Schnitzbauer A.
        • et al.
        The rate of cholangiocarcinoma in Caroli Disease A German multicenter study.
        HPB (Oxford). 2022; 24: 267-276https://doi.org/10.1016/j.hpb.2021.06.423