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Proteomic analysis of X19728 exposure in triple-negative breast cancer (TNBC) cells

ABSTRACT: To define the downstream protein-level consequences of X19728 exposure in triple-negative breast cancer (TNBC) cells, wild-type and AHR-knockout MDA-MB-468 cells were treated with vehicle control (DMSO) or 3 uM X19728 (2-phenyl-imidazo[1,2-a]pyridine) for 36 hours. Following treatment, whole-cell lysates were collected. Proteins were extracted, reduced, alkylated, and digested with trypsin prior to analysis using an Evosep One liquid chromatography system coupled to a timsTOF HT mass spectrometer (Bruker).

INSTRUMENT(S): timsTOF HT

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Siva Kolluri

PROVIDER: MSV000100825 | MassIVE | Thu Feb 12 10:53:00 GMT 2026

REPOSITORIES: MassIVE

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Project description:Purpose:Triple negative breast cancer (TNBC) commonly metastasizes to the brain and predicts poor prognosis with limited therapeutic options. TNBC frequently harbors BRCA mutations translating to platinum sensitivity; platinum response may be augmented by additional suppression of DNA repair mechanisms through poly(ADP-ribose)polymerase (PARP) inhibition. We evaluated brain penetrance and efficacy of Carboplatin +/- the PARP inhibitor ABT888, and investigated gene expression changes in murine intracranial (IC) TNBC models stratified by BRCA and molecular subtype status. Experimental design:Athymic mice were inoculated intra-cerebrally with BRCA-mutant: SUM149 (basal), MDA-MB-436 (claudin-low), or BRCA-wild-type: MDA-MB-468 (basal), MDA-MB-231BR (claudin-low) TNBC cells and treated with PBS control (IP, weekly), Carboplatin (50mg/kg/week, IP), ABT888 (25mg/kg/day, OG), or their combination. DNA-damage (?-H2AX) and apoptosis (cleaved-Caspase-3(cC3)) were assessed via IHC of IC tumors. Gene expression of BRCA-mutant IC tumors was measured. Results: Carboplatin+/-ABT888 significantly improved survival in BRCA-mutant IC models compared to control, but did not improve survival in BRCA-wild-type IC models. Carboplatin+ABT888 revealed a modest survival advantage versus Carboplatin in BRCA-mutant models. ABT888 yielded a marginal survival benefit in the MDA-MB-436 but not in the SUM149 model. BRCA-mutant SUM149 expression of ?-H2AX and cC3 proteins was elevated in all treatment groups compared to Control, while BRCA-wild-type MDA-MB-468 cC3 expression did not increase with treatment. Carboplatin treatment induced common gene expression changes in BRCA-mutant models.Conclusions: Carboplatin+/-ABT888 improves survival in BRCA-mutant IC TNBC models with corresponding DNA damage and gene expression changes. Combination therapy represents a promising treatment strategy for patients with TNBC brain metastases warranting further clinical investigation. reference x sample

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Proteomic analysis of X19728 exposure in triple-negative breast cancer (TNBC) cells

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