Cell lineage tracing links ERα loss in Erbb2-positive breast cancers to the arising of a highly aggressive breast cancer subtype.

Abstract

HER2-positive (HER2⁺) breast cancers (BrCs) contain approximately equal numbers of ERα⁺HER2⁺ and ERα^-HER2⁺ cases. An enduring obstacle is the unclear cell lineage-related characteristics of these BrCs. Although ERα⁺HER2⁺ BrCs could lose ERα to become ERα^-HER2⁺ BrCs, direct evidence is missing. To investigate ERα dependencies and their implications during BrC growth and metastasis, we generated ERα^CreRFP-T mice that produce an RFP-marked ERα⁺ mammary gland epithelial cell (MGEC) lineage. RCAS virus-mediated expression of Erbb2, a rodent Her2 homolog, first produced comparable numbers of ERα⁺RFP⁺Erbb2⁺ and ERα^-RFP^-Erbb2⁺ MGECs. Early hyperplasia developed mostly from ERα⁺RFP⁺Erbb2⁺ cells and ERα^-RFP^-Erbb2⁺ cells in these lesions were rare. The subsequently developed ductal carcinomas in situ had 64% slow-proliferating ERα⁺RFP⁺Erbb2⁺ cells, 15% fast-proliferating ERα^-RFP⁺Erbb2⁺ cells derived from ERα⁺RFP⁺Erbb2⁺ cells, and 20% fast-proliferating ERα^-RFP^-Erbb2⁺ cells. The advanced tumors had mostly ERα^-RFP⁺Erbb2⁺ and ERα^-RFP^-Erbb2⁺ cells and only a very small population of ERα⁺RFP⁺Erbb2⁺ cells. In ERα^-RFP⁺Erbb2⁺ cells, GATA3 and FoxA1 decreased expression and ERα promoter regions became methylated, consistent with the loss of ERα expression. Lung metastases consisted of mostly ERα^-RFP⁺Erbb2⁺ cells, a few ERα^-RFP^-Erbb2⁺ cells, and no ERα⁺RFP⁺Erbb2⁺ cells. The high metastatic capacity of ERα^-RFP⁺Erbb2⁺ cells was associated with ERK1/2 activation. These results show that the slow-proliferating, nonmetastatic ERα⁺RFP⁺Erbb2⁺ cells progressively lose ERα during tumorigenesis to become fast-proliferating, highly metastatic ERα^-RFP⁺Erbb2⁺ cells. The ERα^-Erbb2⁺ BrCs with an ERα⁺ origin are more aggressive than those ERα^-Erbb2⁺ BrCs with an ERα^- origin, and thus, they should be distinguished and treated differently in the future.