Ataxia-telangiectasia (AT) and ataxia-oculomotor apraxia type 2 (AOA2) are both autosomal recessive cerebellar ataxias characterized by elevated serum alpha-fetoprotein (AFP) levels. However, the source and clinical implications of this increase, as well as its relationship with liver diseases are unknown. In this study, we investigated the frequency of liver diseases and their relationship with high AFP in patients with AT and AOA2. The study involved 19 adult patients (13 patients with AT and 6 patients with AOA2) who were followed between January 1992 and March 2023. The demographic and clinical characteristics, serum levels of liver enzymes and AFP, liver imaging, and survival data were retrospectively reviewed. The mean age of patients was 26.8±5.1 years (11 men and 8 women).While 69% (9/13) of AT patients had elevated liver enzymes and 56% (5/9) had abnormal liver imaging, both were normal in all AOA2 patients.Liver enzyme elevation was associated with the presence of comorbid disease (p=0.007), but not with AFP level (p=0.33) in AT patients. Hepatosteatosis was not associated neither with comorbidity (p=0.524) nor AFP level (p=0.905) in this group. During a median follow-up of 17 (1-29) years, 5 AT patients passed away due to cancer (4 patients) and sepsis (1 patient). AFP level was not associated with the occurrence of cancer (p=0.382). This study found a high prevalence of liver disease (69%) in AT, unlike in AOA, independent of AFP levels. Since comorbid diseases, especially cancer, were associated with elevated liver enzymes, adult AT patients with abnormal liver functions should be screened for the development of cancers.

R-loops are RNA-DNA-hybrid-containing nucleic acids with important cellular roles. Deregulation of R-loop dynamics can lead to DNA damage and genome instability1, which has been linked to the action of endonucleases such as XPG2-4. However, the mechanisms and cellular consequences of such processing have remained unclear. Here we identify a new population of RNA-DNA hybrids in the cytoplasm that are R-loop-processing products. When nuclear R-loops were perturbed by depleting the RNA-DNA helicase senataxin (SETX) or the breast cancer gene BRCA1 (refs. 5-7), we observed XPG- and XPF-dependent cytoplasmic hybrid formation. We identify their source as a subset of stable, overlapping nuclear hybrids with a specific nucleotide signature. Cytoplasmic hybrids bind to the pattern recognition receptors cGAS and TLR3 (ref. 8), activating IRF3 and inducing apoptosis. Excised hybrids and an R-loop-induced innate immune response were also observed in SETX-mutated cells from patients with ataxia oculomotor apraxia type 2 (ref. 9) and in BRCA1-mutated cancer cells10. These findings establish RNA-DNA hybrids as immunogenic species that aberrantly accumulate in the cytoplasm after R-loop processing, linking R-loop accumulation to cell death through the innate immune response. Aberrant R-loop processing and subsequent innate immune activation may contribute to many diseases, such as neurodegeneration and cancer.

Ataxia with oculomotor apraxia is a rare neurodegenerative subgroup of diseases with manifestations that include cerebellar ataxia, oculomotor apraxia, extrapyramidal features, and sensorimotor neuropathy. In 2015, ataxia with oculomotor apraxia type 4 was described in 11 Portuguese individuals. The mean age of onset was 4.3 years, with severe extrapyramidal manifestations, neuropathy, rapid progression, and ataxia, being wheelchair-bound during adolescence. The disease is caused by homozygous or compound heterozygous mutations in the PNKP gene. In this case report, we describe two sisters, who were 52- and 58-years-old, with cerebellar dysarthria, oculomotor apraxia, dystonia, and gait ataxia. Two new mutations in the PNKP gene were detected in both sisters, confirming the diagnosis of ataxia with oculomotor apraxia. They were remarkable because they were able to walk unaided during adulthood and had epilepsy. With these clinical cases, we attempt to raise awareness of the possibility of different phenotypes of this rare disease, expanding the spectrum of manifestations of ataxia with oculomotor apraxia type 4.

Premature loss of ovarian activity before 40 years of age is known as primary ovarian insufficiency (POI) and occurs in ∼1% of women. A more subtle decline in ovarian activity, known as premature ovarian ageing (POA), occurs in ∼10% of women. Despite the high prevalence of POA, very little is known regarding its genetic causation. Senataxin (SETX) is an RNA/DNA helicase involved in repair of oxidative stress-induced DNA damage. Homozygous mutation of SETX leads to the neurodegenerative disorder, ataxia oculomotor apraxia type 2 (AOA2). There have been reports of POI in AOA2 females suggesting a link between SETX and ovarian ageing. Here, we studied female mice lacking either one (Setx+/-) or both (Setx-/-) copies of SETX over a 12- to 14-month period. We find that DNA damage is increased in oocytes from 8-month-old Setx+/- and Setx-/- females compared with Setx+/+ oocytes leading to a marked reduction in all classes of ovarian follicles at least 4 months earlier than typically occurs in female mice. Furthermore, during a 12-month long mating trial, Setx+/- and Setx-/- females produced significantly fewer pups than Setx+/+ females from 7 months of age onwards. These data show that SETX is critical for preventing POA in mice, likely by preserving DNA integrity in oocytes. Intriguingly, heterozygous Setx loss causes an equally severe impact on ovarian ageing as homozygous Setx loss. Because heterozygous SETX disruption is less likely to produce systemic effects, SETX compromise could underpin some cases of insidious POA.

Ataxia with oculomotor apraxia type 2 (AOA2) is a slowly progressive, autosomal recessive disease characterized by the triad of ataxia, oculomotor apraxia, and sensorimotor neuropathy. The genetic basis of AOA2 is biallelic mutation of the SETX gene, resulting in reduced or absent senataxin, a DNA/RNA repair protein essential for genomic stability. In this case report, we described a case of AOA2 with two clear pathogenic SETX mutations, one of which is novel. We then discussed two further likely "in cis" SETX sequence changes (previously reported in the literature as pathogenic), and presented the case that they are likely benign polymorphisms.