title: Differentiation of human telencephalic progenitor cells into MSNs by inducible expression of Gsx2 and Ebf1.
abstract: Medium spiny neurons (MSNs) are a key population in the basal ganglia network, and their degeneration causes a severe neurodegenerative disorder, Huntington's disease. Understanding how ventral neuroepithelial progenitors differentiate into MSNs is critical for regenerative medicine to develop specific differentiation protocols using human pluripotent stem cells. Studies performed in murine models have identified some transcriptional determinants, including GS Homeobox 2 (Gsx2) and Early B-cell factor 1 (Ebf1). Here, we have generated human embryonic stem (hES) cell lines inducible for these transcription factors, with the aims of (i) studying their biological role in human neural progenitors and (ii) incorporating TF conditional expression in a developmental-based protocol for generating MSNs from hES cells. Using this approach, we found that Gsx2 delays cell-cycle exit and reduces Pax6 expression, whereas Ebf1 promotes neuronal differentiation. Moreover, we found that Gsx2 and Ebf1 combined overexpression in hES cells achieves high yields of MSNs, expressing Darpp32 and Ctip2, in vitro as well in vivo after transplantation. We show that hES-derived striatal progenitors can be transplanted in animal models and can differentiate and integrate into the host, extending fibers over a long distance.
authors.affiliation: Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122 Milan, Italy; andrea.faedo@unimi.it elena.cattaneo@unimi.it.|Istituto Nazionale Genetica Molecolare (INGM) Romeo ed Enrica Invernizzi, Milan 20122, Italy.
authors.forename: Andrea
authors.identifier: 0000-0002-2227-0656
authors.initials: A
authors.lastname: Faedo
authors.affiliation: Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122 Milan, Italy.|Istituto Nazionale Genetica Molecolare (INGM) Romeo ed Enrica Invernizzi, Milan 20122, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Laporta
authors.affiliation: Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122 Milan, Italy.|Istituto Nazionale Genetica Molecolare (INGM) Romeo ed Enrica Invernizzi, Milan 20122, Italy.
authors.forename: Alice
authors.initials: A
authors.lastname: Segnali
authors.affiliation: Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122 Milan, Italy.|Istituto Nazionale Genetica Molecolare (INGM) Romeo ed Enrica Invernizzi, Milan 20122, Italy.
authors.forename: Maura
authors.initials: M
authors.lastname: Galimberti
authors.affiliation: Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122 Milan, Italy.|Istituto Nazionale Genetica Molecolare (INGM) Romeo ed Enrica Invernizzi, Milan 20122, Italy.
authors.forename: Dario
authors.initials: D
authors.lastname: Besusso
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Cesana
authors.affiliation: Institute of Molecular Bioimaging and Physiology, National Research Council (IBFM-CNR), San Raffaele Scientific Institute, University of Milan Bicocca, 20132 Milan, Italy.|Department of Medicine and Surgery, University of Milan Bicocca, 20126 Milan, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Belloli
authors.affiliation: Institute of Molecular Bioimaging and Physiology, National Research Council (IBFM-CNR), San Raffaele Scientific Institute, University of Milan Bicocca, 20132 Milan, Italy.|Department of Medicine and Surgery, University of Milan Bicocca, 20126 Milan, Italy.
authors.forename: Rosa Maria
authors.initials: RM
authors.lastname: Moresco
authors.affiliation: Department of Neuroscience, Neuroscience Institute Cavalieri Ottolenghi, 10043 Torino, Italy.|Department of Neuroscience, National Institute of Neuroscience, 10043 Torino, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Tropiano
authors.affiliation: Department of Neuroscience, Neuroscience Institute Cavalieri Ottolenghi, 10043 Torino, Italy.|Department of Neuroscience, National Institute of Neuroscience, 10043 Torino, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Fucà
authors.affiliation: Miltenyi Biotec GmbH, 51429 Bergisch Gladbach, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Wild
authors.affiliation: Miltenyi Biotec GmbH, 51429 Bergisch Gladbach, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bosio
authors.affiliation: Department of Neuroscience, Neuroscience Institute Cavalieri Ottolenghi, 10043 Torino, Italy.|Department of Neuroscience, National Institute of Neuroscience, 10043 Torino, Italy.
authors.forename: Alessandro E
authors.initials: AE
authors.lastname: Vercelli
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Biella
authors.affiliation: Laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases, Department of Biosciences, University of Milan, 20122 Milan, Italy; andrea.faedo@unimi.it elena.cattaneo@unimi.it.|Istituto Nazionale Genetica Molecolare (INGM) Romeo ed Enrica Invernizzi, Milan 20122, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Cattaneo
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32 / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000378:metabolism* / Q000637:transplantation; D006801:Humans; D008819:Mice, Nude; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D033581:Stem Cell Transplantation / Q000379:methods; D013687:Telencephalon / Q000166:cytology; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014183:Transplantation, Heterologous; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C479740:BCL11B protein, human; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; C081976:EBF1 protein, human; C491626:Gsx2 protein, human; D018398:Homeodomain Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D025521:Tumor Suppressor Proteins
keywords: Ebf1; Gsx2; HD; MSNs; hES cells