The Renin Angiotensin System (RAS) mediates bifunctional growth regulation in melanoma and is a novel target for therapeutic intervention

Alexander Renziehausen, Hexiao Wang, Bhavya Rao, Lynda Weir, Cristiana Lo Nigro, Laura Lattanzio, Marco Merlano, Antonio Vega-Rioja, Maria del Carmen Fernandez-Carranco, Nabil Hajji, Rubeta Matin, Catherine Harwood, Su Li, Van Ren Sim, Kevin O’Neill, Alan Evans, Alastair Thompson, Peter Szlosarek, Colin Fleming, Justin StebbingCharlotte Proby, Andreas G. Tzakos, Nelofer Syed, Tim Crook

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    Abstract

    Despite emergence of new systemic therapies, metastatic melanoma remains a challenging and often fatal form of skin cancer. The renin-angiotensin system (RAS) is a major physiological regulatory pathway controlling salt-water equilibrium, intravascular volume and blood pressure. Biological effects of the RAS are mediated by the vasoactive hormone angiotensin II (AngII) via two receptor subtypes, AT1R (encoded by AGTR1) and AT2R (encoded by AGTR2). We report decreasing expression and increasing CpG island methylation of AGTR1 in metastatic versus primary melanoma and detection in serum of methylated genomic DNA from the AGTR1 CpG island in metastatic melanoma implying that AGTR1 encodes a tumour suppressor function in melanoma. Consistent with this hypothesis, antagonism of AT1R using losartan or shRNA-mediated knock-down in melanoma cell lines expressing AGTR1 resulted in acquisition of the ability to proliferate in serum-free conditions. Conversely, ectopic expression of AGTR1 in cell lines lacking endogenous expression inhibits proliferation irrespective of the presence of AngII implying a ligand-independent suppressor function for AT1R. Treatment of melanoma cell lines expressing endogenous AT2R with either AngII or the AT2R-selective agonist Y6AII induces proliferation in serum-free conditions whereas the AT2R-specific antagonists PD123319 and EMA401 inhibit melanoma growth and angiogenesis and potentiate inhibitors of BRAF and MEK in cells with BRAF V600 mutations. Our results demonstrate that the RAS has both oncogenic and tumour suppressor functions in melanoma. Pharmacological inhibition of AT2R may provide therapeutic opportunities in melanomas expressing this receptor and AGTR1 CpG island methylation in serum may serve as a novel biomarker of metastatic melanoma.
    Original languageEnglish
    Pages (from-to)2320-2336
    Number of pages17
    JournalOncogene
    Volume38
    Early online date26 Nov 2018
    DOIs
    Publication statusPublished - 28 Mar 2019

    Fingerprint

    Renin-Angiotensin System
    Melanoma
    Growth
    CpG Islands
    Angiotensin II
    Therapeutics
    Serum
    Cell Line
    Methylation
    Angiogenesis Inhibitors
    Losartan
    Mitogen-Activated Protein Kinase Kinases
    Skin Neoplasms
    Small Interfering RNA
    Neoplasms
    Salts
    Biomarkers
    Hormones
    Pharmacology
    Blood Pressure

    Keywords

    • Melanoma
    • Angiotensin
    • Angiotensin receptor
    • epigenetics
    • Humans
    • Amides/pharmacology
    • Renin-Angiotensin System/drug effects
    • Angiotensin II/pharmacology
    • Neoplasm Metastasis
    • Imidazoles/pharmacology
    • Receptor, Angiotensin, Type 2/genetics
    • Antihypertensive Agents/pharmacology
    • Melanoma/genetics
    • Molecular Targeted Therapy/methods
    • Cell Proliferation/drug effects
    • Cells, Cultured
    • Angiotensin-Converting Enzyme Inhibitors/pharmacology
    • Zebrafish
    • Xenograft Model Antitumor Assays
    • Animals
    • DNA Methylation/drug effects
    • Embryo, Nonmammalian
    • Cell Line, Tumor
    • Fumarates/pharmacology
    • Pyridines/pharmacology
    • Receptor, Angiotensin, Type 1/genetics

    Cite this

    Renziehausen, Alexander ; Wang, Hexiao ; Rao, Bhavya ; Weir, Lynda ; Nigro, Cristiana Lo ; Lattanzio, Laura ; Merlano, Marco ; Vega-Rioja, Antonio ; del Carmen Fernandez-Carranco, Maria ; Hajji, Nabil ; Matin, Rubeta ; Harwood, Catherine ; Li, Su ; Sim, Van Ren ; O’Neill, Kevin ; Evans, Alan ; Thompson, Alastair ; Szlosarek, Peter ; Fleming, Colin ; Stebbing, Justin ; Proby, Charlotte ; Tzakos, Andreas G. ; Syed, Nelofer ; Crook, Tim. / The Renin Angiotensin System (RAS) mediates bifunctional growth regulation in melanoma and is a novel target for therapeutic intervention. In: Oncogene. 2019 ; Vol. 38. pp. 2320-2336.
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    abstract = "Despite emergence of new systemic therapies, metastatic melanoma remains a challenging and often fatal form of skin cancer. The renin-angiotensin system (RAS) is a major physiological regulatory pathway controlling salt-water equilibrium, intravascular volume and blood pressure. Biological effects of the RAS are mediated by the vasoactive hormone angiotensin II (AngII) via two receptor subtypes, AT1R (encoded by AGTR1) and AT2R (encoded by AGTR2). We report decreasing expression and increasing CpG island methylation of AGTR1 in metastatic versus primary melanoma and detection in serum of methylated genomic DNA from the AGTR1 CpG island in metastatic melanoma implying that AGTR1 encodes a tumour suppressor function in melanoma. Consistent with this hypothesis, antagonism of AT1R using losartan or shRNA-mediated knock-down in melanoma cell lines expressing AGTR1 resulted in acquisition of the ability to proliferate in serum-free conditions. Conversely, ectopic expression of AGTR1 in cell lines lacking endogenous expression inhibits proliferation irrespective of the presence of AngII implying a ligand-independent suppressor function for AT1R. Treatment of melanoma cell lines expressing endogenous AT2R with either AngII or the AT2R-selective agonist Y6AII induces proliferation in serum-free conditions whereas the AT2R-specific antagonists PD123319 and EMA401 inhibit melanoma growth and angiogenesis and potentiate inhibitors of BRAF and MEK in cells with BRAF V600 mutations. Our results demonstrate that the RAS has both oncogenic and tumour suppressor functions in melanoma. Pharmacological inhibition of AT2R may provide therapeutic opportunities in melanomas expressing this receptor and AGTR1 CpG island methylation in serum may serve as a novel biomarker of metastatic melanoma.",
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    author = "Alexander Renziehausen and Hexiao Wang and Bhavya Rao and Lynda Weir and Nigro, {Cristiana Lo} and Laura Lattanzio and Marco Merlano and Antonio Vega-Rioja and {del Carmen Fernandez-Carranco}, Maria and Nabil Hajji and Rubeta Matin and Catherine Harwood and Su Li and Sim, {Van Ren} and Kevin O’Neill and Alan Evans and Alastair Thompson and Peter Szlosarek and Colin Fleming and Justin Stebbing and Charlotte Proby and Tzakos, {Andreas G.} and Nelofer Syed and Tim Crook",
    note = "Acknowledgements: The work was supported by The Chief Scientific Officer of Scotland, The Anonymous Trust, Tenovus Scotland (Dr Crook), Melanoma Focus (Professor Proby), The Leng Foundation (Professor Proby) and The Brain Tumour Research Campaign (Dr Syed). Antonio Vega-Rioja is under contract Proyectos I+D+I para jovenes investigadores from de Economica y Competitividad (SAF2014-60649-JN) and co-funded by Fondo Europeo de Regional-FEDER. Tim Crook is a Scottish Senior Clinical Fellow in Medical Oncology.",
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    Renziehausen, A, Wang, H, Rao, B, Weir, L, Nigro, CL, Lattanzio, L, Merlano, M, Vega-Rioja, A, del Carmen Fernandez-Carranco, M, Hajji, N, Matin, R, Harwood, C, Li, S, Sim, VR, O’Neill, K, Evans, A, Thompson, A, Szlosarek, P, Fleming, C, Stebbing, J, Proby, C, Tzakos, AG, Syed, N & Crook, T 2019, 'The Renin Angiotensin System (RAS) mediates bifunctional growth regulation in melanoma and is a novel target for therapeutic intervention', Oncogene, vol. 38, pp. 2320-2336. https://doi.org/10.1038/s41388-018-0563-y

    The Renin Angiotensin System (RAS) mediates bifunctional growth regulation in melanoma and is a novel target for therapeutic intervention. / Renziehausen, Alexander; Wang, Hexiao; Rao, Bhavya; Weir, Lynda; Nigro, Cristiana Lo; Lattanzio, Laura; Merlano, Marco; Vega-Rioja, Antonio; del Carmen Fernandez-Carranco, Maria; Hajji, Nabil; Matin, Rubeta; Harwood, Catherine; Li, Su; Sim, Van Ren; O’Neill, Kevin; Evans, Alan; Thompson, Alastair; Szlosarek, Peter; Fleming, Colin; Stebbing, Justin; Proby, Charlotte; Tzakos, Andreas G.; Syed, Nelofer (Lead / Corresponding author); Crook, Tim (Lead / Corresponding author).

    In: Oncogene, Vol. 38, 28.03.2019, p. 2320-2336.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - The Renin Angiotensin System (RAS) mediates bifunctional growth regulation in melanoma and is a novel target for therapeutic intervention

    AU - Renziehausen, Alexander

    AU - Wang, Hexiao

    AU - Rao, Bhavya

    AU - Weir, Lynda

    AU - Nigro, Cristiana Lo

    AU - Lattanzio, Laura

    AU - Merlano, Marco

    AU - Vega-Rioja, Antonio

    AU - del Carmen Fernandez-Carranco, Maria

    AU - Hajji, Nabil

    AU - Matin, Rubeta

    AU - Harwood, Catherine

    AU - Li, Su

    AU - Sim, Van Ren

    AU - O’Neill, Kevin

    AU - Evans, Alan

    AU - Thompson, Alastair

    AU - Szlosarek, Peter

    AU - Fleming, Colin

    AU - Stebbing, Justin

    AU - Proby, Charlotte

    AU - Tzakos, Andreas G.

    AU - Syed, Nelofer

    AU - Crook, Tim

    N1 - Acknowledgements: The work was supported by The Chief Scientific Officer of Scotland, The Anonymous Trust, Tenovus Scotland (Dr Crook), Melanoma Focus (Professor Proby), The Leng Foundation (Professor Proby) and The Brain Tumour Research Campaign (Dr Syed). Antonio Vega-Rioja is under contract Proyectos I+D+I para jovenes investigadores from de Economica y Competitividad (SAF2014-60649-JN) and co-funded by Fondo Europeo de Regional-FEDER. Tim Crook is a Scottish Senior Clinical Fellow in Medical Oncology.

    PY - 2019/3/28

    Y1 - 2019/3/28

    N2 - Despite emergence of new systemic therapies, metastatic melanoma remains a challenging and often fatal form of skin cancer. The renin-angiotensin system (RAS) is a major physiological regulatory pathway controlling salt-water equilibrium, intravascular volume and blood pressure. Biological effects of the RAS are mediated by the vasoactive hormone angiotensin II (AngII) via two receptor subtypes, AT1R (encoded by AGTR1) and AT2R (encoded by AGTR2). We report decreasing expression and increasing CpG island methylation of AGTR1 in metastatic versus primary melanoma and detection in serum of methylated genomic DNA from the AGTR1 CpG island in metastatic melanoma implying that AGTR1 encodes a tumour suppressor function in melanoma. Consistent with this hypothesis, antagonism of AT1R using losartan or shRNA-mediated knock-down in melanoma cell lines expressing AGTR1 resulted in acquisition of the ability to proliferate in serum-free conditions. Conversely, ectopic expression of AGTR1 in cell lines lacking endogenous expression inhibits proliferation irrespective of the presence of AngII implying a ligand-independent suppressor function for AT1R. Treatment of melanoma cell lines expressing endogenous AT2R with either AngII or the AT2R-selective agonist Y6AII induces proliferation in serum-free conditions whereas the AT2R-specific antagonists PD123319 and EMA401 inhibit melanoma growth and angiogenesis and potentiate inhibitors of BRAF and MEK in cells with BRAF V600 mutations. Our results demonstrate that the RAS has both oncogenic and tumour suppressor functions in melanoma. Pharmacological inhibition of AT2R may provide therapeutic opportunities in melanomas expressing this receptor and AGTR1 CpG island methylation in serum may serve as a novel biomarker of metastatic melanoma.

    AB - Despite emergence of new systemic therapies, metastatic melanoma remains a challenging and often fatal form of skin cancer. The renin-angiotensin system (RAS) is a major physiological regulatory pathway controlling salt-water equilibrium, intravascular volume and blood pressure. Biological effects of the RAS are mediated by the vasoactive hormone angiotensin II (AngII) via two receptor subtypes, AT1R (encoded by AGTR1) and AT2R (encoded by AGTR2). We report decreasing expression and increasing CpG island methylation of AGTR1 in metastatic versus primary melanoma and detection in serum of methylated genomic DNA from the AGTR1 CpG island in metastatic melanoma implying that AGTR1 encodes a tumour suppressor function in melanoma. Consistent with this hypothesis, antagonism of AT1R using losartan or shRNA-mediated knock-down in melanoma cell lines expressing AGTR1 resulted in acquisition of the ability to proliferate in serum-free conditions. Conversely, ectopic expression of AGTR1 in cell lines lacking endogenous expression inhibits proliferation irrespective of the presence of AngII implying a ligand-independent suppressor function for AT1R. Treatment of melanoma cell lines expressing endogenous AT2R with either AngII or the AT2R-selective agonist Y6AII induces proliferation in serum-free conditions whereas the AT2R-specific antagonists PD123319 and EMA401 inhibit melanoma growth and angiogenesis and potentiate inhibitors of BRAF and MEK in cells with BRAF V600 mutations. Our results demonstrate that the RAS has both oncogenic and tumour suppressor functions in melanoma. Pharmacological inhibition of AT2R may provide therapeutic opportunities in melanomas expressing this receptor and AGTR1 CpG island methylation in serum may serve as a novel biomarker of metastatic melanoma.

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