Combination therapy for KIT-mutant mast cells: Targeting constitutive NFAT and KIT activity

Alison C. Macleod, Lillian R. Klug, Janice Patterson, Diana J. Griffith, Carol Beadling, Ajia Town, Michael Heinrich

    Research output: Contribution to journalArticle

    4 Citations (Scopus)

    Abstract

    Resistant KIT mutations have hindered the development of KIT kinase inhibitors for treatment of patients with systemic mastocytosis. The goal of this research was to characterize the synergistic effects of a novel combination therapy involving inhibition of KIT and calcineurin phosphatase, a nuclear factor of activated T cells (NFAT) regulator, using a panel of KIT-mutant mast cell lines. The effects of monotherapy or combination therapy on the cellular viability/survival of KIT-mutant mast cells were evaluated. In addition, NFAT-dependent transcriptional activity was monitored in a representative cell line to evaluate the mechanisms responsible for the efficacy of combination therapy. Finally, shRNA was used to stably knockdown calcineurin expression to confirm the role of calcineurin in the observed synergy. The combination of a KIT inhibitor and a calcineurin phosphatase inhibitor (CNPI) synergized to reduce cell viability and induce apoptosis in six distinct KIT-mutant mast cell lines. Both KIT inhibitors and CNPIs were found to decrease NFAT-dependent transcriptional activity. NFAT-specific inhibitors induced similar synergistic apoptosis induction as CNPIs when combined with a KIT inhibitor. Notably, NFAT was constitutively active in each KIT-mutant cell line tested. Knockdown of calcineurin subunit PPP3R1 sensitized cells to KIT inhibition and increased NFAT phosphorylation and cytoplasmic localization. Constitutive activation of NFAT appears to represent a novel and targetable characteristic of KIT-mutant mast cell disease. Our studies suggest that combining KIT inhibition with NFATinhibition might represent a new treatment strategy for mast cell disease.

    Original languageEnglish (US)
    Pages (from-to)2840-2851
    Number of pages12
    JournalMolecular Cancer Therapeutics
    Volume13
    Issue number12
    DOIs
    StatePublished - Dec 1 2014

    Fingerprint

    NFATC Transcription Factors
    Mast Cells
    Calcineurin
    Mastocytosis
    Cell Line
    Therapeutics
    Systemic Mastocytosis
    Apoptosis
    Small Interfering RNA
    Cell Survival
    Phosphotransferases
    Phosphorylation
    Mutation
    Research

    ASJC Scopus subject areas

    • Oncology
    • Cancer Research
    • Medicine(all)

    Cite this

    Combination therapy for KIT-mutant mast cells : Targeting constitutive NFAT and KIT activity. / Macleod, Alison C.; Klug, Lillian R.; Patterson, Janice; Griffith, Diana J.; Beadling, Carol; Town, Ajia; Heinrich, Michael.

    In: Molecular Cancer Therapeutics, Vol. 13, No. 12, 01.12.2014, p. 2840-2851.

    Research output: Contribution to journalArticle

    Macleod, Alison C. ; Klug, Lillian R. ; Patterson, Janice ; Griffith, Diana J. ; Beadling, Carol ; Town, Ajia ; Heinrich, Michael. / Combination therapy for KIT-mutant mast cells : Targeting constitutive NFAT and KIT activity. In: Molecular Cancer Therapeutics. 2014 ; Vol. 13, No. 12. pp. 2840-2851.
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