Preclinical comparison of proteasome and ubiquitin E1 enzyme inhibitors in cutaneous squamous cell carcinoma

the identification of mechanisms of differential sensitivity

Angela McHugh, Kenneth Fernandes, Andrew South, Jemima E. Mellerio, Julio C. Salas-Alanis, Charlotte Proby, Irene Leigh, Mark Saville

    Research output: Contribution to journalArticle

    5 Citations (Scopus)
    67 Downloads (Pure)

    Abstract

    Proteasome inhibitors have distinct properties and the biochemical consequences of suppressing ubiquitin E1 enzymes and the proteasome differ. We compared the effects of the proteasome inhibitors bortezomib, ixazomib and carfilzomib and the ubiquitin E1 enzyme inhibitor MLN7243/TAK-243 on cell viability and cell death in normal keratinocytes and cutaneous squamous cell carcinoma (cSCC) cell lines. The effects of both a pulse of treatment and more extended incubation were investigated. This is relevant to directly-delivered therapy (topical treatment/intratumoral injection) where the time of exposure can be controlled and a short exposure may better reflect systemically-delivered inhibitor pharmacokinetics. These agents can selectively kill cSCC cells but there are variations in the pattern of cSCC cell line sensitivity/resistance. Variations in the responses to proteasome inhibitors are associated with differences in the specificity of the inhibitors for the three proteolytic activities of the proteasome. There is greater selectivity for killing cSCC cells compared to normal keratinocytes with a pulse of proteasome inhibitor treatment than with a more extended exposure. We provide evidence that c-MYC-dependent NOXA upregulation confers susceptibility to a short incubation with proteasome inhibitors by priming cSCC cells for rapid BAK-dependent death. We observed that bortezomibresistant cSCC cells can be sensitive to MLN7243-induced death. Low expression of the ubiquitin E1 UBA1/UBE1 participates in conferring susceptibility to MLN7243 by increasing sensitivity to MLN7243-mediated attenuation of ubiquitination. This study supports further investigation of the potential of proteasome and ubiquitin E1 inhibition for cSCC therapy. Direct delivery of inhibitors could facilitate adequate exposure of skin cancers.

    Original languageEnglish
    Pages (from-to)20265-20281
    Number of pages17
    JournalOncotarget
    Volume9
    DOIs
    Publication statusPublished - 17 Apr 2018

    Fingerprint

    Enzyme Inhibitors
    Proteasome Endopeptidase Complex
    Ubiquitin
    Proteasome Inhibitors
    Squamous Cell Carcinoma
    Skin
    Keratinocytes
    Cell Line
    Ubiquitination
    Skin Neoplasms
    Therapeutics
    Cell- and Tissue-Based Therapy
    Cell Survival
    Cell Death
    Up-Regulation
    Pharmacokinetics
    Injections
    Enzymes

    Keywords

    • MLN7243/TAK-243
    • Proteasome
    • Squamous cell carcinoma
    • UBA1
    • UBA6

    Cite this

    @article{e99e17ddec264d6e83e267a703eb6603,
    title = "Preclinical comparison of proteasome and ubiquitin E1 enzyme inhibitors in cutaneous squamous cell carcinoma: the identification of mechanisms of differential sensitivity",
    abstract = "Proteasome inhibitors have distinct properties and the biochemical consequences of suppressing ubiquitin E1 enzymes and the proteasome differ. We compared the effects of the proteasome inhibitors bortezomib, ixazomib and carfilzomib and the ubiquitin E1 enzyme inhibitor MLN7243/TAK-243 on cell viability and cell death in normal keratinocytes and cutaneous squamous cell carcinoma (cSCC) cell lines. The effects of both a pulse of treatment and more extended incubation were investigated. This is relevant to directly-delivered therapy (topical treatment/intratumoral injection) where the time of exposure can be controlled and a short exposure may better reflect systemically-delivered inhibitor pharmacokinetics. These agents can selectively kill cSCC cells but there are variations in the pattern of cSCC cell line sensitivity/resistance. Variations in the responses to proteasome inhibitors are associated with differences in the specificity of the inhibitors for the three proteolytic activities of the proteasome. There is greater selectivity for killing cSCC cells compared to normal keratinocytes with a pulse of proteasome inhibitor treatment than with a more extended exposure. We provide evidence that c-MYC-dependent NOXA upregulation confers susceptibility to a short incubation with proteasome inhibitors by priming cSCC cells for rapid BAK-dependent death. We observed that bortezomibresistant cSCC cells can be sensitive to MLN7243-induced death. Low expression of the ubiquitin E1 UBA1/UBE1 participates in conferring susceptibility to MLN7243 by increasing sensitivity to MLN7243-mediated attenuation of ubiquitination. This study supports further investigation of the potential of proteasome and ubiquitin E1 inhibition for cSCC therapy. Direct delivery of inhibitors could facilitate adequate exposure of skin cancers.",
    keywords = "MLN7243/TAK-243, Proteasome, Squamous cell carcinoma, UBA1, UBA6",
    author = "Angela McHugh and Kenneth Fernandes and Andrew South and Mellerio, {Jemima E.} and Salas-Alanis, {Julio C.} and Charlotte Proby and Irene Leigh and Mark Saville",
    note = "This study was supported by DEBRA International and funded by DEBRA Austria (Saville-Proby 1). I.M.L and M.K.S were supported by an ERC Advanced Investigator Award (250170, Principal Investigator I.M.L.). C.M.P. and I.M.L. were supported by a Cancer Research UK Programme Grant (A13044).",
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    Preclinical comparison of proteasome and ubiquitin E1 enzyme inhibitors in cutaneous squamous cell carcinoma : the identification of mechanisms of differential sensitivity. / McHugh, Angela; Fernandes, Kenneth; South, Andrew; Mellerio, Jemima E.; Salas-Alanis, Julio C.; Proby, Charlotte; Leigh, Irene; Saville, Mark.

    In: Oncotarget, Vol. 9, 17.04.2018, p. 20265-20281.

    Research output: Contribution to journalArticle

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    T2 - the identification of mechanisms of differential sensitivity

    AU - McHugh, Angela

    AU - Fernandes, Kenneth

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    AU - Mellerio, Jemima E.

    AU - Salas-Alanis, Julio C.

    AU - Proby, Charlotte

    AU - Leigh, Irene

    AU - Saville, Mark

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    AB - Proteasome inhibitors have distinct properties and the biochemical consequences of suppressing ubiquitin E1 enzymes and the proteasome differ. We compared the effects of the proteasome inhibitors bortezomib, ixazomib and carfilzomib and the ubiquitin E1 enzyme inhibitor MLN7243/TAK-243 on cell viability and cell death in normal keratinocytes and cutaneous squamous cell carcinoma (cSCC) cell lines. The effects of both a pulse of treatment and more extended incubation were investigated. This is relevant to directly-delivered therapy (topical treatment/intratumoral injection) where the time of exposure can be controlled and a short exposure may better reflect systemically-delivered inhibitor pharmacokinetics. These agents can selectively kill cSCC cells but there are variations in the pattern of cSCC cell line sensitivity/resistance. Variations in the responses to proteasome inhibitors are associated with differences in the specificity of the inhibitors for the three proteolytic activities of the proteasome. There is greater selectivity for killing cSCC cells compared to normal keratinocytes with a pulse of proteasome inhibitor treatment than with a more extended exposure. We provide evidence that c-MYC-dependent NOXA upregulation confers susceptibility to a short incubation with proteasome inhibitors by priming cSCC cells for rapid BAK-dependent death. We observed that bortezomibresistant cSCC cells can be sensitive to MLN7243-induced death. Low expression of the ubiquitin E1 UBA1/UBE1 participates in conferring susceptibility to MLN7243 by increasing sensitivity to MLN7243-mediated attenuation of ubiquitination. This study supports further investigation of the potential of proteasome and ubiquitin E1 inhibition for cSCC therapy. Direct delivery of inhibitors could facilitate adequate exposure of skin cancers.

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