DNA damage response associated miRNA dysregulation in Ataxia-telangiectasia

Abstract Background Ataxia-telangiectasia (A-T) is a rare autosomal recessive genome-instability disorder due to pathogenic variants in ATM, a central kinase of the DNA double-strand break signaling, checkpoint activation and stress-response pathways. Although microRNAs (miRNAs) regulate DNA damage response (DDR), apoptosis, senescence and cell-cycle control, the stress-responsive behavior of selected ATM/DDR-associated miRNAs remains incompletely characterized in A-T fibroblast models. Methods and Results Four predefined ATM/DDR-associated candidate miRNAs (hsa-miR-34a-5p, hsa-miR-26a-5p, hsa-miR-106b-5p and hsa-miR-20a-5p) were quantified in one ATM-proficient fibroblast control and three A-T fibroblast cell lines both under basal and bleomycin exposed conditions. The relative expression of selected miRNAs was measured by RT-qPCR. miR-34a-5p and miR-26a-5p levels were lower in each of the three bleomycin-treated A-T lines relative to the single bleomycin-treated WT control line. miR-34a-5p and miR-26a-5p levels were lower in bleomycin-treated three A-T lines relative to the single control WT line. Within-cell-line analysis showed significant miR-34a-5p induction after bleomycin in WT, AT648 and AT9607, whereas significant miR-26a-5p induction was observed only in AT648. AT648 showed the broadest within-cell-line response, with significant induction of all four tested miRNAs after bleomycin. Target-based enrichment and network analyses of the predefined candidate panel identified annotations related to cell-cycle regulation, G1/S transition, RB/E2F signaling, p53 signaling, apoptosis, senescence, PI3K-Akt signaling, and DNA damage-associated processes. Network analysis revealed a number of shared candidate targets such as CCND1 , CCND2 , E2F1 , E2F3 , RB1 , WEE1 , PTEN , SMAD4 , MYC and VEGFA . Conclusions In this fibroblast panel, miR-34a-5p and miR-26a-5p showed the most consistent baseline alterations, whereas bleomycin responsiveness was cell-line specific and heterogeneous.