title: Dickkopf 3 Promotes the Differentiation of a Rostrolateral Midbrain Dopaminergic Neuronal Subset In Vivo and from Pluripotent Stem Cells In Vitro in the Mouse.
abstract: Wingless-related MMTV integration site 1 (WNT1)/β-catenin signaling plays a crucial role in the generation of mesodiencephalic dopaminergic (mdDA) neurons, including the substantia nigra pars compacta (SNc) subpopulation that preferentially degenerates in Parkinson's disease (PD). However, the precise functions of WNT1/β-catenin signaling in this context remain unknown. Stem cell-based regenerative (transplantation) therapies for PD have not been implemented widely in the clinical context, among other reasons because of the heterogeneity and incomplete differentiation of the transplanted cells. This might result in tumor formation and poor integration of the transplanted cells into the dopaminergic circuitry of the brain. Dickkopf 3 (DKK3) is a secreted glycoprotein implicated in the modulation of WNT/β-catenin signaling. Using mutant mice, primary ventral midbrain cells, and pluripotent stem cells, we show that DKK3 is necessary and sufficient for the correct differentiation of a rostrolateral mdDA neuron subset. Dkk3 transcription in the murine ventral midbrain coincides with the onset of mdDA neurogenesis and is required for the activation and/or maintenance of LMX1A (LIM homeobox transcription factor 1α) and PITX3 (paired-like homeodomain transcription factor 3) expression in the corresponding mdDA precursor subset, without affecting the proliferation or specification of their progenitors. Notably, the treatment of differentiating pluripotent stem cells with recombinant DKK3 and WNT1 proteins also increases the proportion of mdDA neurons with molecular SNc DA cell characteristics in these cultures. The specific effects of DKK3 on the differentiation of rostrolateral mdDA neurons in the murine ventral midbrain, together with its known prosurvival and anti-tumorigenic properties, make it a good candidate for the improvement of regenerative and neuroprotective strategies in the treatment of PD. Significance statement: We show here that Dickkopf 3 (DKK3), a secreted modulator of WNT (Wingless-related MMTV integration site)/β-catenin signaling, is both necessary and sufficient for the proper differentiation and survival of a rostrolateral (parabrachial pigmented nucleus and dorsomedial substantia nigra pars compacta) mesodiencephalic dopaminergic neuron subset, using Dkk3 mutant mice and murine primary ventral midbrain and pluripotent stem cells. The progressive loss of these dopamine-producing mesodiencephalic neurons is a hallmark of human Parkinson's disease, which can up to now not be halted by clinical treatments of this disease. Thus, the soluble DKK3 protein might be a promising new agent for the improvement of current protocols for the directed differentiation of pluripotent and multipotent stem cells into mesodiencephalic dopaminergic neurons and for the promotion of their survival in situ.
authors.affiliation: Institutes of Developmental Genetics and.
authors.forename: Yoshiyasu
authors.initials: Y
authors.lastname: Fukusumi
authors.affiliation: Institutes of Developmental Genetics and.
authors.forename: Florian
authors.identifier: http://orcid.org/0000-0003-4451-5025
authors.initials: F
authors.lastname: Meier
authors.affiliation: Institutes of Developmental Genetics and.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Götz
authors.affiliation: Institutes of Developmental Genetics and.
authors.forename: Friederike
authors.identifier: http://orcid.org/0000-0002-5947-3203
authors.initials: F
authors.lastname: Matheus
authors.affiliation: Experimental Genetics, Helmholtz Center Munich, German Research Center for Environmental Health, D-85764 Neuherberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Irmler
authors.affiliation: Emil Fischer Centre, Institute of Biochemistry, Friedrich Alexander University Erlangen-Nuremberg, D-91054 Erlangen, Germany.
authors.forename: Ruth
authors.initials: R
authors.lastname: Beckervordersandforth
authors.affiliation: Institutes of Developmental Genetics and.
authors.forename: Theresa
authors.initials: T
authors.lastname: Faus-Kessler
authors.affiliation: Institutes of Developmental Genetics and.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Minina
authors.affiliation: Institutes of Developmental Genetics and.
authors.forename: Benedict
authors.identifier: http://orcid.org/0000-0003-4003-4208
authors.initials: B
authors.lastname: Rauser
authors.affiliation: Institutes of Developmental Genetics and.
authors.forename: Jingzhong
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Laboratory of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics and.
authors.forename: Ernest
authors.initials: E
authors.lastname: Arenas
authors.affiliation: Department of Cell and Molecular Biology, Karolinska Institute, S-17177 Stockholm, Sweden.
authors.forename: Elisabet
authors.identifier: http://orcid.org/0000-0003-3095-465X
authors.initials: E
authors.lastname: Andersson
authors.affiliation: Division of Molecular Embryology, German Cancer Research Center-Center for Molecular Biology at Heidelberg University Alliance, German Cancer Research Center, D-69120 Heidelberg, Germany, Institute of Molecular Biology, D-55128 Mainz, Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: Niehrs
authors.affiliation: Experimental Genetics, Helmholtz Center Munich, German Research Center for Environmental Health, D-85764 Neuherberg, Germany, Technical University Munich, Chair of Experimental Genetics, D-85350 Freising-Weihenstephan, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Beckers
authors.affiliation: Institute of Genetics and Biophysics "A. Buzzati-Traverso," I-80131 Naples, Italy, Institute for Inpatient Treatment and Scientific Studies Neuromed, I-86077 Pozzilli, Isernia, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Simeone
authors.affiliation: Institutes of Developmental Genetics and Technical University Munich, Chair of Developmental Genetics/Helmholtz Center Munich, D-85764 Neuherberg, Germany, German Center for Neurodegenerative Diseases, D-80336 Munich, Germany, and Munich Cluster for Systems Neurology, Adolf Butenandt Institute, Ludwig Maximilians University Munich, D-80336 Munich, Germany wurst@helmholtz-muenchen.de nilima.prakash@hshl.de.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wurst
authors.affiliation: Institutes of Developmental Genetics and Technical University Munich, Chair of Developmental Genetics/Helmholtz Center Munich, D-85764 Neuherberg, Germany, wurst@helmholtz-muenchen.de nilima.prakash@hshl.de.
authors.forename: Nilima
authors.identifier: http://orcid.org/0000-0002-0568-6262
authors.initials: N
authors.lastname: Prakash
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002452:Cell Count; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D003857:Deoxyuridine / Q000031:analogs & derivatives / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000502:physiology*; D008636:Mesencephalon / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D058953:Neural Stem Cells / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000502:physiology*; D059467:Transcriptome; D051155:Wnt1 Protein / Q000235:genetics / Q000502:physiology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C405061:Dkk3 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D051155:Wnt1 Protein; C495246:Wnt1 protein, mouse; C031086:5-ethynyl-2'-deoxyuridine; D003857:Deoxyuridine
keywords: DKK3; WNT1; differentiation; mouse; stem cell; substantia nigra dopamine neuron