Hexokinase 3 enhances myeloid cell survival via non-glycolytic functions

Purpose: The goal was to compare transcriptome profiles of HL60 HK3-null and Cas9 expressing control cells treated with either ATRA or DMSO using RNA sequencing. In addition, to find accessible DNA regions by probing open chromatin with hyperactive mutant Tn5 Transposase using ATAC sequencing. Methods: HL60 HK3-null and Cas9 expressing control cells were treated with 1µM of ATRA or DMSO (0.01%) for 36h. Treated cells were then harvested for RNA-Seq and ATAC-Seq analysis. Results: We mapped sequence reads per sample to the human genome (GRCh38 - hg38) and identified differentially expressed transcripts as well as open chromatin peaks in AML HL60 HK3-null and Cas9 expressing cells upon treatment with ATRA or DMSO as control. Differentially expressed genes, were identified with a fold change ≥1.5 and FDR p-value <0.05. Conclusions: Our results show that HK3 knockout (HK3-null) AML cells had an accumulation of reactive oxygen species (ROS) as well as DNA damage during ATRA-induced differentiation. RNA sequencing analysis confirmed pathway enrichment for programmed cell death, oxidative stress and DNA damage response in HK3-null AML cells. These signatures were confirmed in ATAC sequencing, showing that loss of HK3 leads to changes in chromatin configuration and increases accessibility of genes involved in apoptosis and stress response.