Prelude to malignancy: A gene expression signature in normal mammary gland from breast cancer patients suggests pre‐tumorous alterations and is associated with adverse outcomes

Abstract Despite advances in early detection and treatment strategies, breast cancer recurrence and mortality remain a significant health issue. Recent insights suggest the prognostic potential of microscopically healthy mammary gland, in the vicinity of the breast lesion. Nonetheless, a comprehensive understanding of the gene expression profiles in these tissues and their relationship to patient outcomes remain missing. Furthermore, the increasing trend towards breast‐conserving surgery may inadvertently lead to the retention of existing cancer‐predisposing mutations within the normal mammary gland. This study assessed the transcriptomic profiles of 242 samples from 83 breast cancer patients with unfavorable outcomes, including paired uninvolved mammary gland samples collected at varying distances from primary lesions. As a reference, control samples from 53 mammoplasty individuals without cancer history were studied. A custom panel of 634 genes linked to breast cancer progression and metastasis was employed for expression profiling, followed by whole‐transcriptome verification experiments and statistical analyses to discern molecular signatures and their clinical relevance. A distinct gene expression signature was identified in uninvolved mammary gland samples, featuring key cellular components encoding keratins, CDH1, CDH3, EPCAM cell adhesion proteins, matrix metallopeptidases, oncogenes, tumor suppressors, along with crucial genes ( FOXA1 , RAB25 , NRG1 , SPDEF , TRIM29 , and GABRP ) having dual roles in cancer. Enrichment analyses revealed disruptions in epithelial integrity, cell adhesion, and estrogen signaling. This signature, named KAOS for Keratin‐Adhesion‐Oncogenes‐Suppressors, was significantly associated with reduced tumor size but increased mortality rates. Integrating molecular assessment of non‐malignant mammary tissue into disease management could enhance survival prediction and facilitate personalized patient care.