Open Access Highly Accessed Research article

Effectiveness and molecular interactions of the clinically active mTORC1 inhibitor everolimus in combination with tamoxifen or letrozole in vitro and in vivo

Lesley-Ann Martin1*, Sunil Pancholi1, Ian Farmer1, Stephanie Guest1, Ricardo Ribas1, Marion T Weigel1, Allan M Thornhill2, Zara Ghazoui1, Roger A'Hern3, Dean B Evans4, Heidi A Lane4, Stephen R Johnston5 and Mitch Dowsett15

Author Affiliations

1 Breakthrough Breast Cancer Research Centre, Institute of Cancer Research, London, SW3 6JB UK

2 McElwain Laboratory, Institute of Cancer Research, London, UK

3 Clinical Trial Unit, Institute of Cancer Research, Surrey, SM2 5NG, UK

4 Novartis Institutes for BioMedical Research, CH-4002, Basel, Switzerland

5 The Royal Marsden Hospital, London, SW3 6JB, UK

For all author emails, please log on.

Breast Cancer Research 2012, 14:R132  doi:10.1186/bcr3330

Published: 17 October 2012

Abstract

Introduction

Strategies to improve the efficacy of endocrine agents in breast cancer (BC) therapy and to delay the onset of resistance include concomitant targeting of the estrogen receptor alpha (ER) and the mammalian target of rapamycin complex 1 (mTORC1), which regulate cell-cycle progression and are supported by recent clinical results.

Methods

BC cell lines expressing aromatase (AROM) and modeling endocrine-sensitive (MCF7-AROM1) and human epidermal growth factor receptor 2 (HER2)-dependent de novo resistant disease (BT474-AROM3) and long-term estrogen-deprived (LTED) MCF7 cells that had acquired resistance associated with HER2 overexpression were treated in vitro and as subcutaneous xenografts with everolimus (RAD001-mTORC1 inhibitor), in combination with tamoxifen or letrozole. End points included proliferation, cell-cycle arrest, cell signaling, and effects on ER-mediated transactivation.

Results

Everolimus caused a concentration-dependent decrease in proliferation in all cell lines, which was associated with reductions in S6 phosphorylation. Everolimus plus letrozole or tamoxifen enhanced the antiproliferative effect and G1-accumulation compared with monotherapy, as well as increased phosphorylation (Ser10) and nuclear accumulation of p27 and pronounced dephosphorylation of Rb. Sensitivity was greatest to everolimus in the LTED cells but was reduced by added estrogen. Increased pAKT occurred in all circumstances with everolimus and, in the BT474 and LTED cells, was associated with increased pHER3. Decreased ER transactivation suggested that the effectiveness of everolimus might be partly related to interrupting cross-talk between growth-factor signaling and ER. In MCF7-AROM1 xenografts, letrozole plus everolimus showed a trend toward enhanced tumor regression, versus the single agents. In BT474-AROM3 xenografts, everolimus alone was equally effective at reducing tumor volume as were the combination therapies.

Conclusions

The results provide mechanistic support for recent positive clinical data on the combination of everolimus and endocrine therapy, as well as data on potential routes of escape via enhanced HER2/3 signaling. This merits investigation for further improvements in treatment efficacy.