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

Jonas Walter,Kamil Grzyb,Lisa M Smits,Michèle Studer,Fredrik Hoel,Silvia Bolognin,Xiaobing Qing,Alexander Skupin,Rejko Krüger,Tomasz Ignac,Michele Bertacchi,Emanuel Berger,Anna S Monzel,Thomas Sauter,Lasse Sinkkonen,Javier Jarazo,Jochen Ohnmacht,Luis Salamanca,Stefano Magni,Rita Perfeito,Laura González-Cano ,Paul Antony ,Karl Johan Tronstad ,Luís Pereira De Almeida ,Sarah Louise Nickels ,Suresh Poovathingal ,Déborah Gérard ,Jens Christian Schwamborn

doi:10.1016/j.celrep.2021.109864

Abstract

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 invivo 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.

Highlights

Parkinson’s disease (PD) is the second most prevalent neurodegenerative disorder after Alzheimer’s disease
Using high-throughput image analyses and single-cell RNA sequencing, we observe that the PD-associated leucine-rich repeat serine/threonine-protein kinase 2 (LRRK2)-G2019S mutation alters the initial phase of neuronal differentiation by accelerating cell-cycle exit with a concomitant increase in cell death
The expression of NR2F1, a key transcription factor involved in neurogenesis, decreases in LRRK2-G2019S neuroepithelial stem cells (NESCs), neurons, and midbrain organoids compared to controls

Summary

Introduction

Parkinson’s disease (PD) is the second most prevalent neurodegenerative disorder after Alzheimer’s disease. It is histopathologically characterized by the progressive loss of midbrain dopaminergic neurons (mDAs) in the substantia nigra (Przedborski, 2017; Schapira et al, 2017). The pathology’s wide spectrum of clinical manifestations is only partially explained by the progressive loss of mDAs. Several lines of evidence have substantiated the hypothesis that PD might have a neurodevelopment component. The number of mDAs has been found to be decreased in the substantia nigra but strongly increased in the olfactory bulb (Huisman et al, 2004). LRRK2 is highly expressed during development (Zechel et al, 2010), and it binds to proteins of the dishevelled (DLV) family (Sancho et al, 2009), key mediators of the Wnt signaling

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