Abstract 4037: High throughput kinase inhibitor screen reveals novel inhibitor combinations acting in synergy with <i>TUBB3</i>/βIII-tubulin suppression in non-small cell lung cancer
Authors: Wee Siang Teo, Tanya Dwarte, Tim W. Failes, Greg M. Arndt, Joshua A. McCarroll, Maria Kavallaris
Journal: Cancer Research (2017)

Abstract Background: Non-Small Cell Lung Cancer (NSCLC) survival rates remain dismal and this malignancy is poorly responsive to current therapy. The mechanisms contributing to this resistance phenotype and aggressive behavior of this disease are not well defined. TUBB3/βIII-tubulin is aberrantly expressed in NSCLC and its expression correlates negatively with disease free survival of patients. Our group has previously shown that βIII-tubulin plays a functional role in chemotherapy resistance and tumorigenesis in NSCLC via modulation of the PTEN/AKT signaling pathway1, 2. This study aimed to identify kinase inhibitors that synergize with βIII-tubulin inhibition in NSCLC using a high-throughput cell-based screen. Methods: A high-throughput cell-based screen of a kinase inhibitor library was performed against our previously described1, 2 NSCLC H460 cells, expressing either βIII-tubulin shRNA or control non-targeting shRNA. The primary screen consisted of a total of 210 kinase inhibitors tested at a single, fixed concentration and cell viability determined using the Alamar blue assay. Secondary screens were performed using a five point dose response. Further validation of the kinase inhibitors was performed against two independent NSCLC, H460 and A549 expressing either βIII-tubulin shRNA or control non-targeting shRNA. Results: A number of kinase inhibitors that act on key signalling cascades were identified to act in synergy with TUBB3/βIII-tubulin knockdown to inhibit NSCLC cell growth. Interestingly, several different inhibitors which target mTOR were identified as top candidates for acting in combination with βIII-tubulin knockdown to inhibit NSCLC cell growth. Treatment of cells with clinically approved mTOR inhibitors, Temsirolimus and Rapamycin, resulted in a significant decrease in cell viability in the βIII-tubulin knockdown cells compared to control. Notably, mTOR acts downstream of AKT signalling and further highlights a potential connection with βIII-tubulin and the AKT signal transduction cascade. Other kinase inhibitors that act on key signaling cascades including Amuvatinib-Multi Kinase Inhibitor (Flt3, Met, Kit, c-Ret &amp; PDGFR); Mubritinib-HER-2 inhibitor; and Neratinib HER2, EGFR, also acted in combination with βIII-tubulin knockdown to inhibit NSCLC cell growth. Conclusions: Our results suggest that clinically approved therapies designed to target mTOR combined with inhibition of TUBB3/βIII-tubulin may offer a novel therapeutic strategy for the treatment of NSCLC patients. References: 1 McCarroll et al. Cancer Research 2010; 70: 4995-5003. 2 McCarroll et al. Cancer Research 2015; 75: 415-425. Citation Format: Wee Siang Teo, Tanya Dwarte, Tim W. Failes, Greg M. Arndt, Joshua A. McCarroll, Maria Kavallaris. High throughput kinase inhibitor screen reveals novel inhibitor combinations acting in synergy with TUBB3/βIII-tubulin suppression in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4037. doi:10.1158/1538-7445.AM2017-4037