LAUSR

Elevated tumor necrosis factor (TNF) α serum levels constitute an adverse prognostic factor for the survival of acute myeloid leukemia (AML) patients. TNFα is produced by various immune effectors and directly or indirectly acts as a pro-proliferative autocrine tumor growth factor. TNFα-controlled regulatory mechanisms especially by long non-coding (lnc)RNA largely remain to be investigated. Here we used DNA microarrays, validated by RNA sequencing, to determine most regulated coding and non-coding transcripts after TNFα stimulation of erythroleukemic TF-1 AML cells. We selected the lncRNA PTTG1-1:1, also identified as MIR3142HG or ENST00000517927, as the most upregulated lncRNA. This long intergenic non-coding RNA of 2301 bases comprises two exons, is strongly conserved in higher vertebrates (Online Supplementary Figure S1) and co-expressed with genes associated with the inflammatory response (Online Supplementary Figure S2). It has recently been reported to be downregulated in adrenocortical carcinoma. We investigated its regulation by pro-inflammatory mediators and validated differential expression in AML cell lines and patient samples compared to healthy controls (Online Supplementary Figure S3). We validated the translational impact of expression of PTTG1-1:1 by in silico analysis using BloodSpot BloodPool, The Cancer Genome Atlas (TCGA), Verhaak and Therapeutically Applicable Research to Generate Effective Treatment (TARGET) AML databases. In line with a role in inflammation, the putative promoter region of PTTG1-1:1 surrounding the transcriptional start site (TSS) from -2,000 to +500 base pairs (bp) (Online Supplementary Figure S1) revealed two nuclear factor (NF)kB binding sites within a chromatin area hypersensitive to DNase I characterized by elevated histone (H)3K27 acetylation (Ac) and H3K4 monomethylation (Me1) and trimethylation (Me3) levels, reflecting transactivation potential. None of the predicted peptides (over