Abstract B013: Associations of epigenome-wide DNA methylation patterns with chemosensitivity and chemoresistance of small cell lung cancer cell lines
Authors: Krushkal Julia , Silvers Thomas , Sonkin Dmitriy , Vural Suleyman , Connelly John , Varma Sudhir , Meltzer Paul S. , Reinhold William C. , Rapisarda Annamaria , Evans David , Pommier Yves , Teicher Beverly A. 
Published: March 2020

Abstract
Small cell lung cancer (SCLC), an aggressive neuroendocrine type of lung cancer, rapidly acquires resistance to treatment. SCLC progression, lineage differentiation, and resistance to therapy have been suggested to involve epigenetic processes. To date, epigenetic links connecting SCLC DNA methylation patterns to drug response and the ways in which these links are mediated by gene expression remain unclear. In order to understand how DNA methylation may affect SCLC response to chemotherapy, we performed an epigenome-wide association study of 66 SCLC cell lines. We used Illumina Infinium MethylationEPIC BeadChip to measure methylation of 866,091 probes. We examined how methylation of probes and gene regions was associated with SCLC in vitro response to 526 antitumor agents. We also identified associations of epigenetic variation with drug response which may be mediated by regulation of gene expression. A potentially important strong association was observed for TREX1, which encodes the 3’ exonuclease I (DNase III) that is involved in resolution of chromatin bridges and has a potential role in chromothripsis. Increased methylation and low expression of TREX1 were associated with SCLC cell line sensitivity to multiple Aurora kinase inhibitors AZD-1152, SCH-1473759, SNS-314, and TAK-901, as well as to the CDK inhibitor R-547, Vertex ATR inhibitor Cpd 45, and the mitotic spindle disruptor vinorelbine. TREX1 upregulation has been previously associated with resistance of other cancers to DNA damaging agents and with DNA repair or DNA degradation after drug exposure. In our analysis, when compared to other cancer categories, TREX1 in SCLC cell lines had low mRNA expression and increased DNA methylation upstream of its transcription start site, which may provide a possible molecular mechanism for SCLC sensitivity to Aurora kinase inhibitors. CEP350 and MLPH, which are involved in centrosome machinery and microtubule tracking, were associated with several Aurora kinase inhibitors and other agents. Among other examples, EPAS1 (HIF2A) was associated with several Aurora kinase inhibitors, the PLK1 inhibitor GSK-461364, and the Bcl-2 inhibitor ABT-737. Methylation of KDM1A, encoding the histone modifier lysine demethylase 1A (LSD1), was associated with PLK1 inhibitors and the KSP inhibitor SB-743921. IGFBP5, which is expressed in the tuft cell-like SCLC subtype, was associated with the mTOR inhibitor INK-128. Upstream regions of MDM2 and DLL3, a Notch pathway regulator overexpressed in ASCL1-high SCLC tumors, were associated with Bcl-2 inhibitors. Methylation and expression of YAP1, a SCLC lineage driver regulating the Hippo pathway, were correlated with the MTOR inhibitor rapamycin. Among non-neuroendocrine lineage markers, EPHA2 was associated with Aurora kinase inhibitors and a PLK1 inhibitor, and CD151 with Bcl-2 inhibitors. Increased methylation upstream of SLFN11 was correlated with resistance to DNA damaging agents, which is likely mediated by SLFN11 expression. The 5’ UTR region of the epigenetic modifier EZH2 was associated with Aurora kinase inhibitors and the FGFR inhibitor BGJ-398. These and multiple other associations identified in this study provide a novel understanding of epigenetic mechanisms which may modulate SCLC response to chemotherapy, and suggest potential molecular targets for combination therapies. This research was supported in part with federal funds from the National Cancer Institute, NIH, under contract HHSN261200800001E.
Citation Format: Julia Krushkal, Thomas Silvers, Dmitriy Sonkin, Suleyman Vural, John Connelly, Sudhir Varma, Paul S. Meltzer, William C. Reinhold, Annamaria Rapisarda, David Evans, Yves Pommier, Beverly A. Teicher. Associations of epigenome-wide DNA methylation patterns with chemosensitivity and chemoresistance of small cell lung cancer cell lines [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B013. doi:10.1158/1535-7163.TARG-19-B013