title: Phenotypic high-throughput screening elucidates target pathway in breast cancer stem cell-like cells.
abstract: Cancer stem cells (CSCs) are resistant to standard cancer treatments and are likely responsible for cancer recurrence, but few therapies target this subpopulation. Due to the difficulty in propagating CSCs outside of the tumor environment, previous work identified CSC-like cells by inducing human breast epithelial cells into an epithelial-to-mesenchymal transdifferentiated state (HMLE_sh_ECad). A phenotypic screen was conducted against HMLE_sh_ECad with 300 718 compounds from the Molecular Libraries Small Molecule Repository to identify selective inhibitors of CSC growth. The screen yielded 2244 hits that were evaluated for toxicity and selectivity toward an isogenic control cell line. An acyl hydrazone scaffold emerged as a potent and selective scaffold targeting HMLE_sh_ECad. Fifty-three analogues were acquired and tested; compounds ranged in potency from 790 nM to inactive against HMLE_sh_ECad. Of the analogues, ML239 was best-in-class with an IC(50)= 1.18 µM against HMLE_sh_ECad, demonstrated a >23-fold selectivity over the control line, and was toxic to another CSC-like line, HMLE_shTwist, and a breast carcinoma cell line, MDA-MB-231. Gene expression studies conducted with ML239-treated cells showed altered gene expression in the NF-κB pathway in the HMLE_sh_ECad line but not in the isogenic control line. Future studies will be directed toward the identification of ML239 target(s).
authors.affiliation: Chemical Biology Platform, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. lcarmody@broadinstitute.org
authors.forename: Leigh C
authors.initials: LC
authors.lastname: Carmody
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Germain
authors.forename: Lynn
authors.initials: L
authors.lastname: VerPlank
authors.forename: Partha P
authors.initials: PP
authors.lastname: Nag
authors.forename: Benito
authors.initials: B
authors.lastname: Muñoz
authors.forename: Jose R
authors.initials: JR
authors.lastname: Perez
authors.forename: Michelle A J
authors.initials: MA
authors.lastname: Palmer
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D005260:Female; D020869:Gene Expression Profiling; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D006835:Hydrazones / Q000737:chemistry / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D016328:NF-kappa B / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D011758:Pyrroles / Q000494:pharmacology*; D054852:Small Molecule Libraries / Q000494:pharmacology*
chemical_list: D006835:Hydrazones; C578078:ML239; D016328:NF-kappa B; D011758:Pyrroles; D054852:Small Molecule Libraries