A Tumor Cell-Selective Inhibitor of Mitogen-Activated Protein Kinase Phosphatases Sensitizes Breast Cancer Cells to Lymphokine-Activated Killer Cell Activity.
Dual specificity mitogen-activated protein kinase (MAPK) phosphatases [dual specificity phosphatase/MAP kinase phosphatase (DUSP-MKP)] have been hypothesized to maintain cancer cell survival by buffering excessive MAPK signaling caused by upstream activating oncogenic products. A large and diverse body of literature suggests that genetic depletion of DUSP-MKPs can reduce tumorigenicity, suggesting that hyperactivating MAPK signaling by DUSP-MKP inhibitors could be a novel strategy to selectively affect the transformed phenotype. Through in vivo structure-activity relationship studies in transgenic zebrafish we recently identified a hyperactivator of fibroblast growth factor signaling [(E)-2-benzylidene-5-bromo-3-(cyclohexylamino)-2,3-dihydro-1H-inden-1-one (BCI-215)] that is devoid of developmental toxicity and restores defective MAPK activity caused by overexpression of DUSP1 and DUSP6 in mammalian cells. Here, we hypothesized that BCI-215 could selectively affect survival of transformed cells. In MDA-MB-231 human breast cancer cells, BCI-215 inhibited cell motility, caused apoptosis but not primary necrosis, and sensitized cells to lymphokine-activated killer cell activity. Mechanistically, BCI-215 induced rapid and sustained phosphorylation of extracellular signal-regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK) in the absence of reactive oxygen species, and its toxicity was partially rescued by inhibition of p38 but not JNK or ERK. BCI-215 also hyperactivated MKK4/SEK1, suggesting activation of stress responses. Kinase phosphorylation profiling documented BCI-215 selectively activated MAPKs and their downstream substrates, but not receptor tyrosine kinases, SRC family kinases, AKT, mTOR, or DNA damage pathways. Our findings support the hypothesis that BCI-215 causes selective cancer cell cytotoxicity in part through non-redox-mediated activation of MAPK signaling, and the findings also identify an intersection with immune cell killing that is worthy of further exploration.
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Christof T
CT
Kaltenmeier
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Laura L
LL
Vollmer
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Lawrence A
LA
Vernetti
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Lindsay
L
Caprio
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Keanu
K
Davis
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Vasiliy N
VN
Korotchenko
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Billy W
BW
Day
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Michael
M
Tsang
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Keren I
KI
Hulkower
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.).
Michael T
MT
Lotze
Departments of Surgery, Immunology and Biochemistry (C.T.K., M.T.L.), Drug Discovery Institute (L.L.V., L.A.V., L.C., K.D., M.T.L., A.V.), Department of Computational and Systems Biology (L.A.V., A.V.), Department of Pharmaceutical Sciences (V.N.K., B.W.D.), and Department of Developmental Biology (M.T.), University of Pittsburgh, Pittsburgh, Pennsylvania; and Platypus Technologies, LLC, Madison, Wisconsin (K.I.H.) avogt@pitt.edu.
Andreas
A
Vogt
D000818:Animals; D030801:Animals, Genetically Modified; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use; D001943:Breast Neoplasms / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D005260:Female; D006367:HeLa Cells; D022781:Hepatocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D015979:Killer Cells, Lymphokine-Activated / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D054639:Mitogen-Activated Protein Kinase Phosphatases / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D015027:Zebrafish
D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D048031:JNK Mitogen-Activated Protein Kinases; D054639:Mitogen-Activated Protein Kinase Phosphatases