Maturation State-Specific Alternative Splicing in FLT3-ITD and NPM1 Mutated AML

SIMPLE SUMMARY: In hematological malignancies, genome-wide sequencing studies found the process of splicing to be surprisingly frequently disrupted. While recent studies characterized altered splicing in relation to splicing factor mutations in AML, this study explored differential splicing profiles associated with two most common aberrations in AML: FLT3-ITD and NPM1 mutations. We identified the differential splicing of FAB-type specific gene sets in FLT3-ITD+/NPM1+ specimens as compared to FLT3-ITD−/NPM1− samples. The primary functions perturbed by differential splicing in all three FAB types included cell cycle control and DNA damage response. Interestingly, differential expression mainly affected genes involved in hematopoietic differentiation. Our findings increase our understanding of how genetic mutations translate to phenotypic features of AML cells to further improve response predictions and to find innovative therapeutic approaches. Altogether, to the best of our knowledge, this is the first study to report differential splicing profiles associated with FLT3-ITD with a concomitant NPM1 mutation in AML. ABSTRACT: Despite substantial progress achieved in unraveling the genetics of AML in the past decade, its treatment outcome has not substantially improved. Therefore, it is important to better understand how genetic mutations translate to phenotypic features of AML cells to further improve response predictions and to find innovative therapeutic approaches. In this respect, aberrant splicing is a crucial contributor to the pathogenesis of hematological malignancies. Thus far, altered splicing is well characterized in relation to splicing factor mutations in AML. However, splicing profiles associated with mutations in other genes remain largely unexplored. In this study, we explored differential splicing profiles associated with two of the most common aberrations in AML: FLT3-ITD and NPM1 mutations. Using RNA-sequencing data of a total of 382 primary AML samples, we found that the co-occurrence of FLT3-ITD and mutated NPM1 is associated with differential splicing of FAB-type specific gene sets. Despite the FAB-type specificity of particular gene sets, the primary functions perturbed by differential splicing in all three FAB types include cell cycle control and DNA damage response. Interestingly, we observed functional divergence between alternatively spliced and differentially expressed genes in FLT3-ITD+/NPM1+ samples in all analyzed FAB types, with differential expression affecting genes involved in hematopoietic differentiation. Altogether, these observations indicate that concomitant FLT3-ITD and mutated NPM1 are associated with the maturation state-specific differential splicing of genes with potential oncogenic relevance.