The Parkinson’s-disease-associated mutation LRRK2-G2019S alters dopaminergic differentiation dynamics via NR2F1

Increasing evidence suggests that neurodevelopmental alterations might contribute to increase the susceptibility to develop neurodegenerative diseases. We investigate the occurrence of developmental abnormalities in dopaminergic neurons in a model of Parkinson’s disease (PD). We monitor the differentiation of human patient-specific neuroepithelial stem cells (NESCs) into dopaminergic neurons. Using high-throughput image analyses and single-cell RNA sequencing, we observe that the PD-associated LRRK2-G2019S mutation alters the initial phase of neuronal differentiation by accelerating cell-cycle exit with a concomitant increase in cell death. We identify the NESC-specific core regulatory circuit and a molecular mechanism underlying the observed phenotypes. The expression of NR2F1, a key transcription factor involved in neurogenesis, decreases in LRRK2-G2019S NESCs, neurons, and midbrain organoids compared to controls. We also observe accelerated dopaminergic differentiation in vivo in NR2F1-deficient mouse embryos. This suggests a pathogenic mechanism involving the LRRK2-G2019S mutation, where the dynamics of dopaminergic differentiation are modified via NR2F1. [Display omitted] •LRRK2-G2019S accelerates cell-cycle exit and dopaminergic differentiation•LRRK2-G2019S reduces the viability of differentiating dopaminergic neurons•LRRK2-G2019S downregulates the core regulatory circuit transcription factor NR2F1•Dopaminergic differentiation is accelerated in NR2F1 mutant mouse embryos Walter et al. show that differentiating iPSC-derived neurons carrying the LRRK2-G2019 mutation are characterized by faster initial neuronal differentiation and cell-cycle exit, as well as increased cell death, compared to controls. Downregulation of the transcription factor NR2F1 appears key to mediate LRRK2-G2019S-dependent phenotypes.