Cancer cells induce a set of adaptive response pathways to survive in the face of stressors due to inadequate vascularization1. One such adaptive pathway is the unfolded protein (UPR) or endoplasmic reticulum (ER) stress response mediated in part by the ER-localized transmembrane sensor IRE12 and its substrate XBP13. Previous studies report UPR activation in various human tumors4-6, but XBP1's role in cancer progression in mammary epithelial cells is largely unknown. Triple negative breast cancer (TNBC), a form of breast cancer in which tumor cells do not express the genes for estrogen receptor, progesterone receptor, and Her2/neu, is a highly aggressive malignancy with limited treatment options7, 8. Here, we report that XBP1 is activated in TNBC and plays a pivotal role in the tumorigenicity and progression of this human breast cancer subtype. In breast cancer cell line models, depletion of XBP1 inhibited tumor growth and tumor relapse and reduced the CD44high/CD24low population. Hypoxia-inducing factor (HIF)1α is known to be hyperactivated in TNBCs 9, 10. Genome-wide mapping of the XBP1 transcriptional regulatory network revealed that XBP1 drives TNBC tumorigenicity by assembling a transcriptional complex with HIF1α that regulates the expression of HIF1α targets via the recruitment of RNA polymerase II. Analysis of independent cohorts of patients with TNBC revealed a specific XBP1 gene expression signature that was highly correlated with HIF1α and hypoxia-driven signatures and that strongly associated with poor prognosis. Our findings reveal a key function for the XBP1 branch of the UPR in TNBC and imply that targeting this pathway may offer alternative treatment strategies for this aggressive subtype of breast cancer.

AUTHOR CONTRIBUTIONS G.R. designed the experimental approach, performed the experimental work, analyzed the data, coordinated the project and wrote the manuscript. A.H. performed primary tumour growth and exosome education in vivo studies, cancer cell proliferation in vitro studies, cancer cell culture and exosome isolation, coordinated the project and wrote the manuscript. C.M.K. generated CEMIP overexpression, performed molecular cloning work and genetic manipulation of cancer cells, cancer cell culture and exosome isolation, coordinated the project and wrote the manuscript. I.R.M. performed brain slice ex vivo FACS analysis and exosome education in vivo studies, cancer cell culture and exosome isolation, coordinated the project, wrote and reviewed the manuscript. L.S. performed brain slice ex vivo experimental work, tissue processing and immunostaining, ex vivo and in vivo ImageJ data analysis and quantification, cancer cell invasion in vitro studies, western blot analysis, cancer cell culture and exosome isolation, and contributed to figure panel assembly. D.F. performed density gradient exosome isolation, characterization and analysis, western blot analysis, and cancer cell culture. H.S.K. and P.R.O. performed RNA sequencing data analysis. I.S. performed tissue processing and immunostaining, ex vivo and in vivo ImageJ data analysis and quantification, cancer cell culture and exosome isolation. I.C.S. performed western blot analysis and assisted in analysis of human data.

Adenoid cystic carcinoma of the breast is a rare neoplasm that represents <1% of breast carcinomas. The tumors are histologically indistinguishable from examples in other sites, and they have a generally favorable prognosis. Several studies have investigated the possible correlation between histologic grade in adenoid cystic carcinoma (largely determined by cytology and growth pattern) and prognosis. Some earlier reports concluded that a solid variant of mammary adenoid cystic carcinoma had a more aggressive clinical course, but others did not confirm this impression. This report describes nine patients with a solid variant of mammary adenoid cystic carcinoma that has a striking basaloid appearance. All were women ranging in age from 37 to 83 years. A solitary mass was evident in all patients. Tumor size was 1.1-15 cm (mean 3.7 cm). The tumors exhibited a predominantly solid architecture comprised of basaloid appearing cells with moderate to marked nuclear atypia. Five tumors had >5 mitotic figures per 10 high power microscopic fields. Intercalated ducts were found in all tumors, being well formed in six and poorly formed in three. Immunohistochemical stains for cytokeratins, basement membranes, and vimentin were consistently positive. Surgery was performed in all cases consisting of excision in seven and mastectomy in two. Axillary lymph node metastases were found in two of six axillary dissections and four had negative lymph nodes. The lymph nodes were not examined in three patients. Follow-up information was available for seven patients. Six women had no evidence of disease after follow-up of 2-88 months (mean 32 months), one patient died of unknown causes, and one patient was lost to follow-up. It is concluded that the solid variant of mammary carcinoma with basaloid features is a histologically distinct tumor that is capable of axillary metastases. Long-term follow-up of a larger series of cases will be needed to determine whether the prognosis of these patients differs significantly from that of women with conventional adenoid cystic carcinoma. Presently, these patients are candidates for axillary staging by sentinel lymph node mapping or low axillary dissection if there is no clinical evidence of axillary metastases. Systemic adjuvant treatment would be prudent when axillary nodal metastases are present. Breast-conserving surgery with radiation is an option if negative margins can be achieved because this appears to be a unicentric form of carcinoma.