Alternative splicing in human myeloid cell differentiation

Abstract

Alternative splicing (AS) plays a pivotal role in the differentiation of immune cells and regulates immune responses. Dysregulations in splicing have been linked to abnormal cell function and can result in diseases. As splicing factors (SFs) are critical in AS, this project aims to understand the role of RNA-binding motif protein 47 (RBM47), a SF, in myeloid cell differentiation. Human promyelocytic leukaemia (HL-60) cells were used as a model for monocyte-like and neutrophil-like differentiation. Differentially expressed genes (DEGs) and differential alternative splicing events (DASEs) associated with RBM47 overexpression were identified from RNA sequencing data. RBM47 overexpression was observed to upregulate a large pool of DEGs when these cells were treated with vitamin D3. Six notable DEGs were selected for further experiments. Monocyte-differentiated HL-60 with overexpressed RBM47 had increased ROS production and surface expression of myeloid maturation and monocytic markers. DASEs were validated in three genes. Findings from this project suggest that RBM47 overexpression supports myeloid cell differentiation in HL-60, resulting in more mature myeloid phenotypes, though more work is required before conclusions can be drawn.