Transcriptomic analysis of WT and Polg D257A/D257A bone marrow derived macrophages (BMDMs) challenged with Pseudomonas aeruginosa
Mitochondrial diseases (MtD) represent a significant public health challenge due to their heterogenous clinical presentation, often severe and progressive symptoms, and lack of effective therapies. Environmental exposures, such bacterial and viral infection, can further compromise mitochondrial function and exacerbate the progression of MtD. Infections in MtD patients more frequently progress to sepsis, pneumonia, and other detrimental inflammatory endpoints. However, the underlying immune alterations that enhance immunopathology in MtD remain unclear, constituting a key gap in knowledge that complicates treatment and increases mortality in this vulnerable population. This dataset reports transcriptomic changes in bone marrow derived macrophages isolated from wild type (WT) C57BL/6J mice and a mouse model of polymerase gamma (Polg)-related MtD (Polg D257A/D257A), both at rest and six hours after infection with Pseudomonas aeruginosa strain PAO1. These data reveal a hyperinflammatory innate immune status in Polg D257A macrophages characterized by elevated expression of genes involved in interferon and inflammatory cell death pathways.  This work sheds new light on innate immune dysregulation in a model of mitochondrial dysfunction and reveals potential targets for limiting infection- and inflammation-related complications in Polg-related MtD. Overall design: After 7 days of differentiation of mouse bone marrow progenitor cells into bone marrow-derived macrophages (BMDMs), macrophages were plated and challenged with GFP-expressing Pseudomonas aeruginosa strain O1 (PAO1) for six hours. Total cellular RNA from WT and Polg D257A/D257A BMDMs was prepared using the Quick-RNA microprep kit (Zymo Research) and subjected to Illumina Stranded mRNA library preparation and RNA sequencing at the Experimental Genomics Core of the Texas A&M Institute for Genome Sciences and Society. FASTQ files were normalized, processed, and analyzed using BaseSpace Sequence Hub (Illumina). In brief, the STAR algorithm of RNAseq Alignment V2.0.0 software was utilized to align the results to the Mus musculus/mm10 (RefSeq) reference genome, then RNA-seq Differential Expression V1.0.0 software was used to obtain differential gene expression values and determine statistically significant changes in Polg D257A/D257A BMDMs relative to WT.