Kinase inhibitors of HER2/AKT pathway induce ERK phosphorylation via a FOXO-dependent feedback loop

Inhibitors of the HER2/PI3K/AKT pathway are being developed, and shown promise in clinical trials for various types of cancers. However, development of drug resistance is a challenging problem for therapy. Elucidating various adaptive pathways leading to resistance or reduced sensitivity to drugs targeting the HER2/PI3K/AKT pathway may provide new insights into countering the resistance. Epidermal growth factor receptor (EGFR, aka HER1), which can dimerize with HER2, can activate a cascade consisting of Ras/RAF/MEK/ERK, promoting tumorigenesis. Lapatinib inhibits the kinase activity of both HER1 and HER2. In the current study, we found that repeated treatment of HER2+ breast cancer cells with HER1/2 inhibitor Lapatinib led to increased phosphorylation of RAF, MEK, and ERK, while suppressing HER1 phosphorylation and reduced the active form of Ras, indicating existence of factor(s) activating RAF/MEK/ERK by bypassing RAS activation. Notably, the Lapatinib treatment-induced phosphorylation of ERK was dependent on FOXO transcription factors, which are also activated by Lapatinib-mediated suppression of AKT. Moreover, the Lapatinib-induced phosphorylation of RAF and ERK is inhibited by a pan-PKC inhibitor. Furthermore, the Lapatinib induced increased ERK phosphorylation is correlated with increased stability of c-Myc, which is known to be stabilized by ERK-mediated phosphorylation. Together, these results suggest that chronic inhibition of the HER1/2 by Lapatinib triggers a feedback loop to activate RAF/MEK/ERK pathway, in a FOXO dependent but Ras-independent manner.