Epigenetic downregulation of HER2 during EMT leads to tumor resistance to HER2-targeted therapies in breast cancer

Abstract

HER2 receptor tyrosine kinase (encoded by ERBB2 gene) is overexpressed in approximately 25% of all breast cancer tumors (known as HER2-positive breast cancers). Overexpression of HER2 causes overactivation of downstream receptor tyrosine kinase pathways including PI3K/Akt and MAPK pathways and is a poor prognosis factor in breast cancer. Tyrosine kinase inhibitor lapatinib and anti-HER2 monoclonal antibodies trastuzumab and pertuzumab are FDA-approved HER2-targeted drugs for treatment of HER2-positive breast cancers. However, development of de novo resistance to HER2 blockade occurs in majority of patients after treatment started. Resistance to HER2 targeting therapies partially due to the loss of HER2 expression on their tumor cells during the treatment. But little is known about the exact mechanism of loss of HER2 on originally HER2-positive tumor cells. Downregulation of extracellular HER2 by metalloproteinases during epithelial-mesenchymal transition (EMT) in trastuzumabresistant/lapatinib-sensitive cells has been shown by limited studies, however, the mechanism of ERBB2 gene silencing during EMT and in the mesenchymal-like cells derived from trastuzumab-resistant/lapatinibresistant HER2-positive breast tumors was entirely unknown. In this study, hypothesized that EMT abrogates HER2 expression by chromatin-based epigenetic silencing of ERBB2 gene as a mechanism of acquired resistance to HER2-targeted therapies. we found that HER2 expression is positively and negatively correlated with the expression of epithelial and mesenchymal phenotype marker genes respectively in breast cancer tumors. We also found that chromatin of ERBB2 gene in HER2-high epithelial-like breast cancer cells is active, while, the chromatin is inactive in HER2-low mesenchymal-like cells. HER2-low breast cancer cell line also revealed less promoter-enhancer interaction and small chromatin loops compared to the HER2-high cell lines. The lower HER2 expression, the higher EMT phenotype, and inactivated chromatin all were found correlated with a lower response to lapatinib. The higher EMT phenotype was found correlated with a lower response to lapatinib. We also found that induction of EMT of HER2-positive breast cancer BT474 cells results in downregulated HER2 expression and lower binding rate of trastuzumab to the cells. These results show that the downregulation of HER2 in mesenchymal-like cells in the culture of HER2-positive breast cancer cell lines was due to ERBB2 gene silencing by epigenetic reprogramming of the cells during EMT. These results indicate that ERBB2 gene silencing by epigenetic regulation during EMT is the main mechanism of resistance of HER2-positive breast cancer cells to trastuzumab and lapatinib.

The relationship between Her 2 neu and N-cadherin expression in hepatocellular carcinoma of Egyptian patients: Immunohistochemical study.

“…The results of the current study showed a highly statistical significant association between high score and strong intensity of expression of Her 2 neu and low score and mild intensity of expression of N-cadherin. These results agree with those of Nami et al, (2020) who detected negative correlation between EMT markers and Her 2 neu expression in breast carcinoma.…”

Assessing the use of tumor-specific DARPin-toxin fusion proteins for ex vivo purging of cancer metastases from human ovarian cortex before autotransplantation

“…These cell lines represent various types of ductal carcinoma (BT-474, HCC202, HCC1954, and MCF7) and adenocarcinoma (MDA-MB-231) and were selected on the basis of their expression levels of EpCAM and HER2. The BT-474, HCC202, and HCC1954 cell lines are all EpCAM and HER2 positive (35,36). The MCF7 cell line is EpCAM-positive, with a low HER2 expression, and the MDA-MB-231 cell line is both EpCAM and HER2 negative (35,37).…”

“…This indicates that in spite of its effectiveness in the elimination of cancer cells, EpCAMtargeted DARPin is not a useful tool to eliminate breast cancer cells from ovarian cortex tissue. We therefore focused on HER2, a transmembrane protein frequently overexpressed in breast cancer cells (35,36). The HER2 protein is not expressed at detectable levels in ovarian cortex tissue (49) and is therefore a potential target for purging with an HER2-targeted DARPin fused to ETA.…”