title: Single-Cell Analysis Reveals a Close Relationship between Differentiating Dopamine and Subthalamic Nucleus Neuronal Lineages.
abstract: Stem cell engineering and grafting of mesencephalic dopamine (mesDA) neurons is a promising strategy for brain repair in Parkinson's disease (PD). Refinement of differentiation protocols to optimize this approach will require deeper understanding of mesDA neuron development. Here, we studied this process using transcriptome-wide single-cell RNA sequencing of mouse neural progenitors expressing the mesDA neuron determinant Lmx1a. This approach resolved the differentiation of mesDA and neighboring neuronal lineages and revealed a remarkably close relationship between developing mesDA and subthalamic nucleus (STN) neurons, while also highlighting a distinct transcription factor set that can distinguish between them. While previous hESC mesDA differentiation protocols have relied on markers that are shared between the two lineages, we found that application of these highlighted markers can help to refine current stem cell engineering protocols, increasing the proportion of appropriately patterned mesDA progenitors. Our results, therefore, have important implications for cell replacement therapy in PD.
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden; Department of Cell and Molecular Biology, Karolinska Institutet, 171 77 Stockholm, Sweden. Electronic address: nigel.kee@licr.ki.se.
authors.forename: Nigel
authors.initials: N
authors.lastname: Kee
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Volakakis
authors.affiliation: Department of Experimental Medical Science, Lund Stem Cell Centre, Lund University, 221 84 Lund, Sweden.
authors.forename: Agnete
authors.initials: A
authors.lastname: Kirkeby
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden.
authors.forename: Lina
authors.initials: L
authors.lastname: Dahl
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden.
authors.forename: Helena
authors.initials: H
authors.lastname: Storvall
authors.affiliation: Department of Experimental Medical Science, Lund Stem Cell Centre, Lund University, 221 84 Lund, Sweden.
authors.forename: Sara
authors.initials: S
authors.lastname: Nolbrant
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden.
authors.forename: Laura
authors.initials: L
authors.lastname: Lahti
authors.affiliation: Department for Cell and Molecular Biology, Science for Life Laboratory, Uppsala University, 751 24 Uppsala, Sweden.
authors.forename: Åsa K
authors.initials: ÅK
authors.lastname: Björklund
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden.
authors.forename: Linda
authors.initials: L
authors.lastname: Gillberg
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden.
authors.forename: Eliza
authors.initials: E
authors.lastname: Joodmardi
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden; Department of Cell and Molecular Biology, Karolinska Institutet, 171 77 Stockholm, Sweden.
authors.forename: Rickard
authors.initials: R
authors.lastname: Sandberg
authors.affiliation: Department of Experimental Medical Science, Lund Stem Cell Centre, Lund University, 221 84 Lund, Sweden.
authors.forename: Malin
authors.initials: M
authors.lastname: Parmar
authors.affiliation: Ludwig Institute for Cancer Research, Box 240, 171 77 Stockholm, Sweden; Department of Cell and Molecular Biology, Karolinska Institutet, 171 77 Stockholm, Sweden. Electronic address: thomas.perlmann@ki.se.
authors.forename: Thomas
authors.initials: T
authors.lastname: Perlmann
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D019521:Body Patterning / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D020869:Gene Expression Profiling; D053263:Gene Regulatory Networks; D006801:Humans; D007150:Immunohistochemistry; D060850:LIM-Homeodomain Proteins / Q000378:metabolism; D055495:Neurogenesis / Q000235:genetics; D017423:Sequence Analysis, RNA; D015398:Signal Transduction / Q000235:genetics; D059010:Single-Cell Analysis / Q000379:methods*; D020531:Subthalamic Nucleus / Q000166:cytology*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D015415:Biomarkers; D060850:LIM-Homeodomain Proteins; C489692:Lmx1a protein, mouse; D014157:Transcription Factors
keywords: Parkinson’s disease; development; dopamine; embryogenesis; graph clustering; hypothalamus; neurogenesis; single cell RNA-seq; stem cell; subthalamic nucleus