Abstract PO1-25-11: Deciphering biological differences between normal breast tissues in men and women using single cell sequencing and in vitro modeling

Abstract Breast cancer in men is relatively rare but outcome from the disease is worse than in women. Although 87% breast cancers in men are Estrogen Receptor-positive (ER+), anti-estrogen therapy is less efficacious in men compared to women with similar breast cancer subtype. Recent genomic analysis of breast tumors in men and women have identified overlapping as well as unique genomic aberrations in breast tumors of men. It is unclear whether genomic aberrations enriched in breast cancers of men uniquely affect the biology and thus alter response to antiestrogen therapy. This lack of knowledge is mainly due to scarcity of model systems. To overcome this limitation, we collected ultrasound guided breast biopsies of healthy men and characterized breast epithelial cells grown from these biopsies. Biopsies were also subjected to single nuclei sequencing. Single nuclei breast atlas of men is currently being superimposed on single nuclei breast atlas generated from breast biopsies of healthy women to determine whether distinct cell types exist in men and to distinguish ductal and lobular breast epithelial cells. Breast epithelial cells of men expressed higher levels of ER compared to similarly propagated breast epithelial cells of women. Expression of ER in a subpopulation of cultured breast epithelial cells was confirmed by immunofluorescence. Among the pioneer factors that determine genome wide binding pattern of ER, TBX3 is expressed at a higher level but FOXA1 is expressed at a lower level in breast epithelial cells of men compared to women. Comparative RNA-seq analysis of breast epithelial cells of men and women revealed elevated non-genomic ER signaling in men compared to women, which could be responsible for limited response of breast tumors of men to anti-estrogen therapy. Additional pathways uniquely upregulated in breast epithelial cells of men include non-canonical Wnt, HIF1A, JUN, NF-kB, TGFb, and FOXO1 signaling. Expectedly, breast epithelial cells of men expressed 15 Y chromosome associated genes including the epigenetic regulators KDM5D and UTY, which have recently been shown to be responsible for sex-specific differences in outcome from other cancer types and loss of Y chromosome could be driver event in certain cancer types. We have created immortalized and transformed variants of breast epithelial cells of men to further aid in mechanistic investigation and drug discovery. Furthermore, our tissue bank currently has breast biopsies of ~30 healthy men to be used as controls for breast cancer studies in men. To our knowledge, this is the first model system to study breast cancer in men starting with breast tissues of healthy men. Citation Format: Poornima Bhat-Nakshatri, Benjamin Fischer, Aditi Khatpe, Adedeji Adebayo, Hongyu Gao, Yunlong Liu, Michele Cote, Harikrishna Nakshatri. Deciphering biological differences between normal breast tissues in men and women using single cell sequencing and in vitro modeling [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-25-11.