The C9orf72 repeat size correlates with onset age of disease, DNA methylation and transcriptional downregulation of the promoter.

Pathological expansion of a G4C2 repeat, located in the 5' regulatory region of C9orf72, is the most common genetic cause of frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). C9orf72 patients have highly variable onset ages suggesting the presence of modifying factors and/or anticipation. We studied 72 Belgian index patients with FTLD, FTLD-ALS or ALS and 61 relatives with a C9orf72 repeat expansion. We assessed the effect of G4C2 expansion size on onset age, the role of anticipation and the effect of repeat size on methylation and C9orf72 promoter activity. G4C2 expansion sizes varied in blood between 45 and over 2100 repeat units with short expansions (45-78 units) present in 5.6% of 72 index patients with an expansion. Short expansions co-segregated with disease in two families. The subject with a short expansion in blood but an indication of mosaicism in brain showed the same pathology as those with a long expansion. Further, we provided evidence for an association of G4C2 expansion size with onset age (P<0.05) most likely explained by an association of methylation state of the 5' flanking CpG island and expansion size in blood (P<0.0001) and brain (P<0.05). In several informative C9orf72 parent-child transmissions, we identified earlier onset ages, increasing expansion sizes and/or increasing methylation states (P=0.0034) of the 5' CpG island, reminiscent of disease anticipation. Also, intermediate repeats (7-24 units) showed a slightly higher methylation degree (P<0.0001) and a decrease of C9orf72 promoter activity (P<0.0001) compared with normal short repeats (2-6 units). Decrease of transcriptional activity was even more prominent in the presence of small deletions flanking G4C2 (P<0.0001). Here we showed that increased methylation of CpGs in the C9orf72 promoter may explain how an increasing G4C2 size lead to loss-of-function without excluding repeat length-dependent toxic gain-of-function. These data provide insights into disease mechanisms and have important implications for diagnostic counseling and potential therapeutic approaches.

Male subfertility: communication, care, coping. An explorative study.

AIM OF THE STUDY: With this study, we wanted to examine the needs of men with fertility problems in terms of communication, care and coping with the diagnosis. METHODOLOGY: data gathered prospectively by means of a written questionnaire (quantitative data), and semi-structured interviews (qualitative data). SAMPLE: 78 subfertile men who consulted for subfertility at the department for reproductive medicine at the University Hospital of Ghent, Belgium were included in the assessment; of these, 23 were interviewed for qualitative evaluation. RESULTS AND CONCLUSIONS: More than one fifth of the participants was dissatisfied with the way they had been informed about their fertility status. There was no significant difference in satisfaction with the care received immediately after diagnosis whether it was given by a general practitioner or by a specialist. A significant influence of nationality was noted on the satisfaction about being informed, Dutch men being much less satisfied than Flemish men. Some men suggested to have a consult with the doctor on a structural basis about a week after the diagnosis. The internet seemed to be a good medium for obtaining medical information IT COULD BE USEFUL TO CREATE AN EXTRA FUNCTION: a 'coach' supporting the couple throughout the entire process., adding another argument to the need for professional psychological support of patients attending clinics for human reproduction. The subfertile men often felt that they were watching from the sideline, and wanted to be more actively involved in the treatment.