Elucidating the importance and regulation of key enhancers for human MEIS1 expression

Myeloid ecotropic virus insertion site 1 (MEIS1) is essential for normal hematopoiesis and is a critical factor in the pathogenesis of a large subset of acute myeloid leukemia (AML). Despite the clinical relevance of MEIS1, its regulation is largely unknown. To understand the transcriptional regulatory mechanisms contributing to human MEIS1 expression, we created a knock-in green florescent protein (GFP) reporter system at the endogenous MEIS1 locus in a human AML cell line. Using this model, we have delineated and dissected a critical enhancer region of the MEIS1 locus for transcription factor (TF) binding through in silico prediction in combination with oligo pull-down, mass-spectrometry and knockout analysis leading to the identification of FLI1, an E-twenty-six (ETS) transcription factor, as an important regulator of MEIS1 transcription. We further show direct binding of FLI1 to the MEIS1 locus in human AML cell lines as well as enrichment of histone acetylation in MEIS1-high healthy and leukemic cells. We also observe a positive correlation between high FLI1 transcript levels and worse overall survival in AML patients. Our study expands the role of ETS factors in AML and our model constitutes a feasible tool for a more detailed understanding of transcriptional regulatory elements and their interactome.

Automated summary

In this study, we created a knock-in GFP reporter system at the endogenous MEIS1 locus in a human AML cell line to investigate the transcriptional regulatory mechanisms of MEIS1. We identified a critical enhancer region of the MEIS1 locus and found FLI1, an ETS transcription factor, to be an important regulator of MEIS1 transcription. Furthermore, we demonstrated direct binding of FLI1 to the MEIS1 locus in human AML cell lines and observed enrichment of histone acetylation in MEIS1-high healthy and leukemic cells. Additionally, high FLI1 transcript levels were associated with worse overall survival in AML patients.