Characterization of mitochondrial alterations in Aicardi-Goutières patients mutated in RNASEH2A and RNASEH2B Genes

The database includes the raw data of the article “Characterization of mitochondrial alterations in Aicardi-Goutières patients mutated in RNASEH2A and RNASEH2B genes”. Aim of the work was to investigate the role of mitochondria in AGS, both from a functional and a morphological point of view in lymphoblastoid cell lines (LCLs) derived from AGS patients mutated in the RNASEH2A and RNASEH2B genes, to ultimately establish the role of mitochondria in the pathogenesis of AGS. The database contains data obtained by means the following evaluations: i) flow cytometry analysis for the evaluation of the intermembrane mitochondrial potential (TMRE-Mitotracker) and for the quantification of produced ROS; ii) Seahorse Analysis for the evaluation of metabolic and energetic status of mutated LCLs; iii) RT-qPCR to determine the expression level of two glycolytic genes (PDK1 and LDHb) and quantify the mtDNA release into the cytoplasm; iiii) Western Blot analysis to quantify Oxphos system complexes protein level.

Both mutant LCLs showed morphological and structural changes in mitochondria when compared to healthy controls, while RNASEH2A LCLs had the most loss of physiological membrane potential. Both mutant LCLs produced more ROS, but RNASEH2B LCLs produced substantially more. RNASEH2B LCLs had a considerably higher amount of cytoplasmic mtDNA. Mitochondrial damage was further confirmed by metabolic changes in both mutant cell lines. In particular RNASEH2A mutated LCLs showed higher ATP production rate and proton leak. These data were also confirmed by the expression level of two glycolytic genes that resulted enhanced in the RNASEH2A mutated cell line. Oxphos system complexes protein level did not show any significant differences. Our findings reveal for the first time the presence of mitochondrial abnormalities in AGS patients' LCLs, implying that these organelles play a critical role in AGS etiology.