Gene expression analysis of multiple myeloma cell lines treated with novel PIM2 inhibitor JP11646

The PIM kinase family (PIM1, 2 and 3) play a central role in integrating growth and survival signals, and are expressed in a wide range of solid and hematological malignancies. We now confirm that PIM2 is overexpressed in multiple myeloma (MM) patients, and within MM group it is overexpressed in the high-risk MF subset (activation of proto-oncogenes MAF/MAFB). This is consistent with our finding of PIM2’s role in key signaling pathways (IL-6, CD28 activation) that confer chemotherapy resistance in MM cells. These studies have identified a novel PIM2-selective non-ATP competitive inhibitor (JP11646) that has a 4 to 760-fold greater suppression of MM proliferation and viability than ATP-competitive PIM inhibitors. This increased efficacy is due not only to the inhibition of PIM2 kinase activity, but also to a novel mechanism involving specific downregulation of PIM2 mRNA and protein expression not seen with the ATP competitive inhibitors. To better understand Off target effects of JP11646 and specific genes that have been directly or indirectly affected by JP11646, MM cell lines MM.1S and U266 were treated with JP11646 and then gene expression was analyzed. The chemotherapeutic sensitive multiple myeloma cell line MM1.S and the more resistant U266 were treated with the ATP non-competitive PIM2 specific novel inhibitor JP11646 (Jasco Pharmaceuticals LLC) for over 24 hours or left untreated and then analyzed for gene expression levels. Cells (5 million per replicate) were treated in triplicate with 20 nM or 200 nM JP11646 (sublethal concentrations that was predetermined to induce 50% cell death over 48 hr and >80% viability over 24 hrs) for 24 hours, or treated in plain media (RPMI1640+10%FBS). Cells were isolated, washed in cold PBS and the RNA isolated was used for gene expression analysis on a Illumina Human HT12 V4 gene chip at the Genomics facility at Roswell Park Cancer Institute, Buffalo, NY. Gene expression values for each gene (average of all the probesets) was normalized, quantile transformed and background corrected and is provided in this submission.