Re-Analysis of GSE275437 Human MDA-MB-231 Breast Adenocarcinoma Cells Treated with 10Gy Radiation

Abstract

Original study investigated how ultrahighdose FLASH radiotherapy can spare normal tissues while controlling tumors by exploring iron homeostasis and ferroptosis. The reanalysis was motivated by the need to deepen mechanistic insight into the transcriptional response to a 10Gy dose in MDAMB231 breast cancer cells, using publicly available RNAseq data to generate a robust gene signature. The workflow employed the ARCHS4 platform to retrieve counts and metadata for GSE275437, constructed an AnnData object and applied Lim- maVoom for differential expression, then leveraged Enrichr to assess enrichment against GO, KEGG, ChEA and mouse phenotype libraries, and used PerturbSeqr to identify smallmolecule and gene perturbations that recapitulate or oppose the signature. The reanalysis revealed that the upregulated set is dominated by a mitotic and tissueremodeling program driven by FOXM1, ESR1/2, MYBL1 and RUNX1 targets, enriched for spindleattachment and focaladhesion disassembly pathways. Conversely, the downregulated set shows coordinated suppression of DNAreplication and repair processes, reflected by E2Ffamily and VDR depletion and enrichment for mismatchrepair pathways. These results underscore a shift from proliferative and DNAre- pair capacity toward heightened mitosis in response to highdose radiation, providing new biomarkers and therapeutic angles for optimizing FLASH radiotherapy. Additionally, perturbation analysis identified kinase inhibitors such as sorafenib and histone deacetylase inhibitors like panobinostat as top mimickers, reinforcing the relevance of cell cycle regulation.