LONA lncRNA overexpression delays myeloid differentiation of OCI-AML3 cell line
Here, we report the identification of one long noncoding RNA (lncRNA) overexpressed in NPM1-mutated AML patients (named LONA) and whose intracellular localization inversely reflects that of NPM1. LONA overexpression in mutant NPM1 cells inhibits in vitro myeloid differentiation and enhances in vivo tumor growth. LONA overexpression combined with a genome-wide RNA-seq search followed by RT-qPCR validations identified a set of mRNA targets encoding proteins involved in myeloid differentiation (including THSB1, MAFB and ASB2) and in tumor cell apoptosis and interaction with its microenvironment. Overall design: We cloned the entire sequence of LONA lncRNA in the retroviral pMSCV-PIG vector. The vector contains a RNA polymerase II promoter driving the lncRNA expression and the PGK promoter driving the puromycin followed by an internal ribosome entry site and GFP . OCI-AML3 (NPM1c+) cell line was transduced with either the PIG-LONA or the PIG EMPTY vector viruses and sorted for GFP+ cells. RNA-sequencing was performed on two independent transductions.