Comprehensive Gene Expression and Epigenetic Profiling of Human Mast Cells: Effects of Butyrate Treatment

This study delves into gene expression and epigenetic modifications in human peripheral blood mononuclear cell-derived mast cells (PBCMCs) with a focus on the effects of butyrate treatment. PBCMCs were generated from CD34+ precursor cells isolated from healthy donors, cultured under serum-free conditions, and tested for maturity. RNA-Seq analysis was conducted to assess differentially expressed genes post-butyrate treatment, revealing significant transcriptional changes. Chromatin Immunoprecipitation Sequencing (ChIP-Seq) was used to explore histone modifications, focusing on H3K27Ac and H3K4Me2 marks. Mature human mast cells were treated with 5 mM butyrate or vehicle for 1, 3, 12 or 24 hours. RNA-Seq was performed to assess differentially expressed genes via Illumina HiSeq 4000, with reads aligned using HISAT, quantified and normalized through HOMER, and differential expression analyzed using DESeq2, followed by pathway enrichment analysis with Metascape. Chromatin Immunoprecipitation Sequencing (ChIP-Seq) was conducted on crosslinked and sonicated chromatin using H3K27Ac and H3K4Me2 antibodies, with sequencing on Illumina HiSeq2500. Data processing involved alignment with HISAT2, peak identification using HOMER, and differential enrichment analysis using DESeq2.