Epithelial to mesenchymal transition is associated with rapamycin resistance

// Ashley M. Holder 1, 2 , Argun Akcakanat 3 , Farrell Adkins 4 , Kurt Evans 3 , Huiqin Chen 5 , Caimiao Wei 5 , Denai R. Milton 5 , Yisheng Li 5 , Kim-Anh Do 5 , Filip Janku 3 , Funda Meric-Bernstam 1, 3 1 Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA 2 Department of Surgery, Washington University in St. Louis, St. Louis, MO, USA 3 Department of Investigational Cancer Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA 4 Department of Colorectal Surgery, Cleveland Clinic Florida, Weston, FL, USA 5 Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA Correspondence to: Funda Meric-Bernstam, e-mail: fmeric@mdanderson.org Keywords: epithelial-to-mesenchymal transition (EMT), rapamycin, trametinib, biomarker, E-cadherin Received: September 02, 2014 Accepted: March 25, 2015 Published: April 13, 2015 ABSTRACT Rapamycin analogues have antitumor efficacy in several tumor types, however few patients demonstrate tumor regression. Thus, there is a pressing need for markers of intrinsic response/resistance and rational combination therapies. We hypothesized that epithelial-to-mesenchymal transition (EMT) confers rapamycin resistance. We found that the epithelial marker E-cadherin protein is higher in rapamycin sensitive (RS) cells and mesenchymal breast cancer cell lines selected by transcriptional EMT signatures are less sensitive to rapamycin. MCF7 cells, transfected with constitutively active mutant Snail, had increased rapamycin resistance (RR) compared to cells transfected with wild-type Snail. Conversely, we transfected two RR mesenchymal cell lines—ACHN and MDA-MB-231—with miR-200b/c or ZEB1 siRNA to promote mesenchymal-to-epithelial transition. This induced E-cadherin expression in both cell lines, and ACHN demonstrated a significant increase in RS. Treatment of ACHN and MDA-MB-231 with trametinib modulated EMT in ACHN cells in vitro . Treatment of MDA-MB-231 and ACHN xenografts with trametinib in combination with rapamycin resulted in significant growth inhibition in both but without an apparent effect on EMT. Future studies are needed to determine whether EMT status is predictive of sensitivity to rapalogs and to determine whether combination therapy with EMT modulating agents can enhance antitumor effects of PI3K/mTOR inhibitors.