Abstract P3-06-03: Dual p38/NLK kinase inhibitor as potential novel therapeutic agent for tamoxifen-resistant luminal breast cancer
Authors: Wang X , Cao X , Veeraraghavan J , Qin L , Kim J-A , Tan Y , Hilsenbeck SG , Schiff R , Wang X 
Published: February 2018

Background: Tamoxifen is the most commonly used endocrine agent for estrogen receptor (ER) positive breast cancer (also known as luminal breast cancer). However, approximately half of the patients develop resistance after initial response to tamoxifen. To date, no effective targeted therapy exists to overcome it. We previously identified the role of nemo-like kinase (NLK), a serine-threonine kinase that functions in stress response and neurite outgrowth, in breast endocrine resistance. In addition, activation of p38 MAPK has been reported to modulate ER signaling and promote endocrine resistance. We identified a highly selective dual p38 and NLK kinase inhibitor (PNKI) through analysis of public kinase profiling datasets, and evaluated its therapeutic effect in endocrine-resistant breast cancers using in vitro and in preclinical mouse models. Experimental design and methods: To determine the in vitro therapeutic window of PNKI, we treated an acquired tamoxifen-resistant cell line (MCF7-TamR) and a benign breast epithelial cell line (MCF10A) with gradually increasing doses of PNKI. To determine the effect of PNKI on tamoxifen-resistant breast cancer cells, we treated primary tamoxifen-resistant breast cancer cell line BT483, and MDAMB415, together with acquired tamoxifen-resistant line MCF7 TamR, T47D TamR, and ZR-75-B TamR, with 0.5 uM PNKI in the presence of different doses of Tamoxifen. To evaluate the therapeutic effect of PNKI in a T47D-derived xenograft tumor model with acquired tamoxifen resistance, we administered PNKI alone or in combination with Fulvestrant, the second-line endocrine therapy agent, or with Everolimus, the mTOR inhibitor that could improve patient outcomes in several clinical trials. Mice bearing xenografts were randomized into six treatment groups (Vehicle, PNKI, Fulvestrant, Fulvestrant+PNKI, Everolimus, Everolimus+PNKI). Tumor growth was tracked closely. The tumors harvested two weeks after treatments started were profiled with Reverse Phase Protein Array (RPPA) to assess the early signaling changes after treatments. The therapeutic effect of PNKI were also evaluated in a patient-derived xenograft (PDX) model of de novo endocrine resistant breast cancer. Mice bearing the PDX tumors were randomized to four treatment groups (Vehicle, PNKI, Everolimus, Everolimus+PNKI) and tumor growth curve was measured timely. Results: PNKI showed an in vitro therapeutic window at 0.1-1μM for MCF7-TamR cells. Breast cancer cell lines with either de novo or acquired Tamoxifen resistance became more sensitive to tamoxifen when treated with 0.5uM PNKI. The concomitant treatment of PNKI and Everolimus results in significant decreased tumor burden and prolonged progression free survival in the both T47D-TamR xenograft tumors and re-transplanted de novo endocrine-resistant PDX tumors compared to other treatments. RPPA data of T47D-TamR tumors harvested following 2-week treatments revealed that several key survival signaling in breast cancer are repressed only when PNKI are combined with Everolimus. Conclusion: The dual p38 and NLK inhibitor (PNKI) exhibited potential therapeutic value as adjuvant agent to the mTOR inhibitor everolimus for acquired or de novo tamoxifen-resistant luminal breast cancers.

Citation Format: Wang X, Cao X, Veeraraghavan J, Qin L, Kim J-A, Tan Y, Hilsenbeck SG, Schiff R, Wang X. Dual p38/NLK kinase inhibitor as potential novel therapeutic agent for tamoxifen-resistant luminal breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P3-06-03.