Growth and early characterization of neural organoids derived from patients with a neurodevelopmental and progeroid disease

Cockayne syndrome (CS) is a rare genetic disease characterized by precocious aging, neurodegeneration, and hypersensitivity to ultraviolet-(UV) radiation, with the brain being one of the most affected organs. This disease is difficult to study, due to the lack or poor representativity of animal models, and the elusive genotype-phenotype correlation concerning the severity of the disease. Cerebral organoids (COs) generated from patient cells reprogrammed into induced pluripotent stem cells (iPSCs) may provide a good experimental model to study molecular defects in CS, in particular neural dysfunctions. COs recapitulate the cytoarchitecture and cell composition of the early developing human brain, including neural rosettes that structure neural stem cells. Previous data in the lab showed that neural rosettes are altered, or absent in the most severe cases, in CS COs. I have generated and analyzed, mainly by immunofluorescence, COs from various CS patients and healthy controls at different time points. Results indicate that CS COs do not form proper neural rosettes, even as early as 16 days, discarding the hypothesis that in CS these structures form and disappear prematurely. I also observed increased cell-to-cell interactions in CS neural rosettes. Apparent reversion of major CS defects was occasionally observed, suggesting compensatory mutations during culturing.