Wnt Inhibitor Screen Reveals Iron Dependence of -Catenin Signaling in Cancers

Department of Biochemistry, Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada.
Cancer Research (Impact Factor: 9.33). 12/2011; 71(24):7628-39. DOI: 10.1158/0008-5472.CAN-11-2745
Source: PubMed

ABSTRACT Excessive signaling from the Wnt pathway is associated with numerous human cancers. Using a high throughput screen designed to detect inhibitors of Wnt/β-catenin signaling, we identified a series of acyl hydrazones that act downstream of the β-catenin destruction complex to inhibit both Wnt-induced and cancer-associated constitutive Wnt signaling via destabilization of β-catenin. We found that these acyl hydrazones bind iron in vitro and in intact cells and that chelating activity is required to abrogate Wnt signaling and block the growth of colorectal cancer cell lines with constitutive Wnt signaling. In addition, we found that multiple iron chelators, desferrioxamine, deferasirox, and ciclopirox olamine similarly blocked Wnt signaling and cell growth. Moreover, in patients with AML administered ciclopirox olamine, we observed decreased expression of the Wnt target gene AXIN2 in leukemic cells. The novel class of acyl hydrazones would thus be prime candidates for further development as chemotherapeutic agents. Taken together, our results reveal a critical requirement for iron in Wnt signaling and they show that iron chelation serves as an effective mechanism to inhibit Wnt signaling in humans.

21 Reads
  • Source
    • "This effect may partly be related to the dependency of the aberrantly activated, tumor-promoting Wnt/␤-catenin pathway on iron excess. On the other hand, Wnt/␤-catenin signaling has been shown to be required for the development of a highly proliferative leukemic stem cell, necessary to maintain leukemias and high levels of ␤-catenin expression correlate with poor prognosis in AML [3]. Hence, the iron chelator DFO might inhibit the leukemic cell proliferation, not only through the reduction of the iron overload along with the decreased expression of NF-␬Bp65 and survivin and increased expression of the apoptotic proteins Bax and Caspase-3, as reported by Yu et al., but also through the abrogation of Wnt/␤catenin signaling. "
    Leukemia Research 09/2014; 38(9). DOI:10.1016/j.leukres.2014.06.021 · 2.35 Impact Factor
  • Source
    • "Iron is an essential element involved in multiple key processes including DNA and heme synthesis, Wnt signalling, and cellular metabolism [4,5]. Many cancer cells exhibit an increased demand for iron in order to maintain their high cellular turnover and DNA synthesis. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Despite improvements in treatment strategies for head and neck squamous cell carcinoma (HNSCC), outcomes have not significantly improved; highlighting the importance of identifying novel therapeutic approaches to target this disease. To address this challenge, we proceeded to evaluate the role of iron in HNSCC. Expression levels of iron-related genes were evaluated in HNSCC cell lines using quantitative RT-PCR. Cellular phenotypic effects were assessed using viability (MTS), clonogenic survival, BrdU, and tumor formation assays. The prognostic significance of iron-related proteins was determined using immunohistochemistry. In a panel of HNSCC cell lines, hemochromatosis (HFE) was one of the most overexpressed genes involved in iron regulation. In vitro knockdown of HFE in HNSCC cell lines significantly decreased hepcidin (HAMP) expression and intracellular iron level. This in turn, resulted in a significant decrease in HNSCC cell viability, clonogenicity, DNA synthesis, and Wnt signalling. These cellular changes were reversed by re-introducing iron back into HNSCC cells after HFE knockdown, indicating that iron was mediating this phenotype. Concordantly, treating HNSCC cells with an iron chelator, ciclopirox olamine (CPX), significantly reduced viability and clonogenic survival. Finally, patients with high HFE expression experienced a reduced survival compared to patients with low HFE expression. Our data identify HFE as potentially novel prognostic marker in HNSCC that promotes tumour progression via HAMP and elevated intracellular iron levels, leading to increased cellular proliferation and tumour formation. Hence, these findings suggest that iron chelators might have a therapeutic role in HNSCC management.
    PLoS ONE 08/2013; 8(8):e74075. DOI:10.1371/journal.pone.0074075 · 3.23 Impact Factor
  • Source
    • "Growth inhibition of test compounds was determined using the Sulforhodamine B (SRB) assay. Cells were plated overnight in 96-well dishes (2500 cells/well) prior to compound or DMSO addition and 72 h later were fixed with 10% (w/v) trichloroacetic acid and stained as published [39]. The amount of SRB present in each well was determined by optical density reading at 490 nm and the results were expressed as a percentage of control cell growth. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Constitutive Wnt signalling is characterized by excessive levels of β-catenin protein and is a frequent occurrence in cancer. APC and Axin are key components of the β-catenin destruction complex that acts to promote β-catenin degradation. The levels of Axin are in turn controlled by tankyrases, members of the PARP-family of poly-ADP-ribosylation enzymes. In colorectal cancer cells, which typically harbor APC mutations, inhibition of tankyrase activity promotes Axin stabilization and attenuates Wnt signalling. Here, we examined the effect of inhibiting tankyrases in breast cancer cells with normal APC. We show that application of the small molecule tankyrase inhibitor, XAV939 or siRNA-mediated abrogation of tankyrase expression increases Axin1 and Axin2 protein levels and attenuates Wnt-induced transcriptional responses in several breast cancer lines. In MDA-MB-231 cells, inhibiton of tankyrase activity also attenuate Wnt3a induced cell migration. Moreover, in both MDA-MB-231 and colorectal cancer cells, XAV939 inhibits cell growth under conditions of serum-deprivation. However, the presence of serum prevents this growth inhibitory effect, although inhibition of Wnt-induced transcriptional and migratory responses was maintained. These results indicate that stabilization of Axin by inhibition of tankyrases alone, may not be an effective means to block tumor cell growth and that combinatorial therapeutic approaches should be considered.
    PLoS ONE 11/2012; 7(11):e48670. DOI:10.1371/journal.pone.0048670 · 3.23 Impact Factor
Show more