Adaptation to mTOR kinase inhibitors by amplification of eIF4E to maintain cap-dependent translation

Abstract

The mechanistic target of rapamycin (mTOR) protein kinase coordinates responses to nutrients and growth factors and is an anti-cancer drug target. To anticipate how cells will respond and adapt to chronic mTORC1 and mTORC2 inhibition we have generated SW620 colon cancer cells with acquired resistance to the ATP-competitive mTOR kinase inhibitor AZD8055 (SW620:8055R). AZD8055 inhibited mTORC1 and mTORC2 signalling and caused a switch from cap-dependent to IRES-dependent translation in parental SW620 cells. In contrast, SW620:8055R cells exhibited a loss of S6K signalling, an increase in expression of the eukaryotic translation initiation factor eIF4E and increased cap-dependent mRNA translation. As a result the expression of CCND1 and MCL1, proteins encoded by eIF4E-sensitive and capdependent transcripts, was refractory to AZD8055 in SW620:8055R cells. RNAimediated knockdown of eIF4E reversed acquired resistance to AZD8055 in SW620:8055R cells; furthermore increased expression of eIF4E was sufficient to reduce sensitivity to AZD8055 in a heterologous cell system. Finally, whilst the combination of MEK1/2 inhibitors with mTOR inhibitors is an attractive rational drug combination, SW620:8055R cells were actually cross-resistant to the MEK1/2 inhibitor selumetinib. These results exemplify the convergence of ERK1/2 and mTOR signalling at eIF4E and the key role of eIF4E downstream of mTOR in maintaining cell proliferation. They also have important implications for therapeutic strategies based around mTOR and MEK1/2-ERK1/2.