title: Transcriptional Mechanisms of Proneural Factors and REST in Regulating Neuronal Reprogramming of Astrocytes.
abstract: Direct lineage reprogramming induces dramatic shifts in cellular identity, employing poorly understood mechanisms. Recently, we demonstrated that expression of Neurog2 or Ascl1 in postnatal mouse astrocytes generates glutamatergic or GABAergic neurons. Here, we take advantage of this model to study dynamics of neuronal cell fate acquisition at the transcriptional level. We found that Neurog2 and Ascl1 rapidly elicited distinct neurogenic programs with only a small subset of shared target genes. Within this subset, only NeuroD4 could by itself induce neuronal reprogramming in both mouse and human astrocytes, while co-expression with Insm1 was required for glutamatergic maturation. Cultured astrocytes gradually became refractory to reprogramming, in part by the repressor REST preventing Neurog2 from binding to the NeuroD4 promoter. Notably, in astrocytes refractory to Neurog2 activation, the underlying neurogenic program remained amenable to reprogramming by exogenous NeuroD4. Our findings support a model of temporal hierarchy for cell fate change during neuronal reprogramming.
authors.affiliation: Physiological Genomics, Biomedical Center, University of Munich, 80336 Munich, Germany; Institute for Stem Cell Research, Helmholtz Centre Munich, 85764 Neuherberg, Germany.
authors.forename: Giacomo
authors.initials: G
authors.lastname: Masserdotti
authors.affiliation: The Francis Crick Institute, Mill Hill Laboratory, The Ridgeway, London, NW7 1AA, UK; Hutchison/MRC Research Center, University of Cambridge, Cambridge Biomedical Campus, Cambridge, CB2 0XZ, UK.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Gillotin
authors.affiliation: Physiological Genomics, Biomedical Center, University of Munich, 80336 Munich, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Sutor
authors.affiliation: The Francis Crick Institute, Mill Hill Laboratory, The Ridgeway, London, NW7 1AA, UK.
authors.forename: Daniela
authors.initials: D
authors.lastname: Drechsel
authors.affiliation: Institute of Experimental Genetics, Helmholtz Centre Munich, 85764 Neuherberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Irmler
authors.affiliation: Department of Medicine, University of Cambridge, Cambridge, CB2 0QQ, UK.
authors.forename: Helle F
authors.initials: HF
authors.lastname: Jørgensen
authors.affiliation: Institute of Computational Biology, Helmholtz Centre Munich, 85764 Neuherberg, Germany.
authors.forename: Steffen
authors.initials: S
authors.lastname: Sass
authors.affiliation: Institute of Computational Biology, Helmholtz Centre Munich, 85764 Neuherberg, Germany; Department of Mathematics, Technical University Munich, 85748 Garching, Germany.
authors.forename: Fabian J
authors.initials: FJ
authors.lastname: Theis
authors.affiliation: Institute of Experimental Genetics, Helmholtz Centre Munich, 85764 Neuherberg, Germany; Center of Life and Food Sciences Weihenstephan, Technical University, 85354 Freising, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Beckers
authors.affiliation: Institute of Physiological Chemistry, University Medical Center of the Johannes Gutenberg University, D-55128 Mainz, Germany; Focus Program Translational Neuroscience, Johannes Gutenberg University, D-55128 Mainz, Germany.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Berninger
authors.affiliation: The Francis Crick Institute, Mill Hill Laboratory, The Ridgeway, London, NW7 1AA, UK. Electronic address: francois.guillemot@crick.ac.uk.
authors.forename: François
authors.initials: F
authors.lastname: Guillemot
authors.affiliation: Physiological Genomics, Biomedical Center, University of Munich, 80336 Munich, Germany; Institute for Stem Cell Research, Helmholtz Centre Munich, 85764 Neuherberg, Germany; Munich Cluster for Systems Neurology "SyNergy," Ludwig Maximilian University of Munich, 80539 Munich, Germany. Electronic address: magdalena.goetz@helmholtz-muenchen.de.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Götz
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism*; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D000099229:RE1-Silencing Transcription Factor
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D004268:DNA-Binding Proteins; D009419:Nerve Tissue Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D000099229:RE1-Silencing Transcription Factor; C499317:Ascl1 protein, mouse; C477059:Insm1 protein, mouse; C104516:Neurod4 protein, mouse; C102938:Neurog2 protein, mouse