Abstract
BACKGROUND AND OBJECTIVES: ADP-ribosylation is a post-translational modification critical for DNA repair, chromatin remodeling, and cellular stress responses. The enzyme ARH3/ADPRHL2 (encoded by the ADPRS gene) is a member of the ADP-ribosyl-acceptor hydrolase family and plays a pivotal role in the removal of mono-ADP-ribosylation, particularly in response to DNA damage. Variants in the ADPRS gene cause stress-induced childhood-onset neurodegeneration with variable ataxia and seizures (CONDSIAS). Here, we present the case of an 11-year-old Brazilian girl with compound heterozygous variants in ADPRS, including a novel missense variant, c.485T>C; p.Leu162Pro, of unknown clinical significance. This study aimed to determine the pathogenicity of this variant and to test the molecular rationale of using minocycline as a therapeutic strategy for CONDSIAS cases. METHODS: Clinical identification and diagnosis of the patient was based primarily on MRI and whole exome sequencing. Functional assays were performed in patient fibroblast cell lines using Western blotting and immunofluorescence-based assays and compared with an ARH3 KO cell line generated by CRISPR/Cas9 technology. RESULTS: Our data indicate that the novel p.Leu162Pro variant, combined with a known nonsense variant, c.316C>T; p.Gln106*, causes a severe reduction of ARH3 protein levels in patient fibroblasts. This leads to defective ARH3-dependent removal of ADP-ribosylation, both under basal conditions and following oxidative stress. Our results using minocycline indicate that it is a poor PARP1 inhibitor and was ineffective in reducing basal ADP-ribosylation in ARH3-deficient cells. DISCUSSION: This study contributes to the growing body of knowledge on ARH3 deficiency associated with CONDSIAS, by identifying a new pathogenic variant and questioning the molecular rationale underlying the therapeutic use of minocycline in patients with CONDSIAS.