Abstract 4206: EUD-GK-001 is a novel kinase inhibitor with in vitro anti-lymphoma activity

Introduction. EUD-GK compounds, and in particular EUD-GK-91, show selectivity for a restricted number of kinases (ALK, RET, TRK, DDR2, ROS1) and have shown anti-tumor activity in specific solid tumor models (Zuccotto et al, EORTC-NCI-AACR 2016). Here, we have assessed a novel derivative of this class of compounds, EUD-GK-001, in a panel of lymphoma models. Methods. Lymphoma cell lines, including both B- and T-cells lymphomas, were exposed to a large range of concentrations of EUD-GK-001 as single agent for 72h, followed by MTT proliferation assay and IC50 calculation. Cell cycle were assessed in four cell lines (FARAGE, DOHH2, TMD8, OCI-LY-10) treated with 500 nM of EUD-GK-001 for 72h. Baseline protein expression was assessed by mass spectrometry. Baseline gene expression was obtained by using the Illumina HumanHT 12 Expression BeadChips. Results. EUD-GK-001 presented a median IC50 of 1.65 μM and 3.75 μM across 14 B- and 4 T- cell lines, respectively. Five out of 14 DLBCL cells lines, including both ABC- and GCB- subtypes, presented IC50 values lower than 1 μM, A dose-depended increase of the cells in the sub-G0 phase (15-55%) was observed in two GCB-DLBCL and two ABC-DLBCL cell lines after EUD-GK-001 exposure for 72h. Taking advantage of available omics data for the tested cell lines (Cascione at al, AACR 2019), functional annotation of RNA and protein levels showed that TNF/NFKB, inflammation, IL6/JAK/STAT3, IL2/STAT5 signatures were enriched among the transcripts more expressed in the sensitive cell lines, while E2F, DNA repair, MYC signatures were enriched in the resistant cell lines. No genes coding for kinases were more expressed in both GCB (n.=2) and ABC (n=2) sensitive than in resistant DLBCL cell lines (GCB, n=2; ABC, n=3 R). However, three kinases, namely SRC, JAK2 and LCK, were significantly more expressed in EUD-GK-001 GCB-DLBCL sensitive cell lines (DOHH2, FARAGE) than in EUD-GK-001 resistant cells (SU-DHL-8, TOLEDO) and merging the top-50 mRNA and top-50 proteins identified by limma, SRC appeared as a central hub of the top genes associated with sensitivity to EUD-GK-001. The pattern of activity of EUD-GK-001 was however different from what reported for the SRC inhibitor dasatinib in the same lymphoma models (Scuoppo et al, PNAS 2019). Conclusion. EUD-GK-001 is a novel small molecule with in vitro anti-tumor activity in subsets of lymphoma models, which needs to be further explored. Citation Format: Lorenzo Scalise, Chiara Tarantelli, Fabio Zuccotto, Filippo Spriano, Michele Mascia, Luciano Cascione, Eugenio Gaudio, Mauro Angiolini, Francesco Bertoni. EUD-GK-001 is a novel kinase inhibitor with in vitro anti-lymphoma activity [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4206.