title: Generation of Dopamine Neurons from Rodent Fibroblasts through the Expandable Neural Precursor Cell Stage.
abstract: Recent groundbreaking work has demonstrated that combined expression of the transcription factors Brn2, Ascl1, and Myt1L (BAM; also known as Wernig factors) convert mouse fibroblasts into postmitotic neuronal cells. However, questions remain regarding whether trans-conversion is achieved directly or involves an intermediary precursor stage. Trans-conversion toward expandable neural precursor cells (NPCs) is more useful than direct one-step neuron formation with respect to yielding a sufficient number of cells and the feasibility of manipulating NPC differentiation toward certain neuron subtypes. Here, we show that co-expression of Wernig factors and Bcl-xL induces fibroblast conversion into NPCs (induced NPCs (iNPCs)) that are highly expandable for >100 passages. Gene expression analyses showed that the iNPCs exhibited high expression of common NPC genes but not genes specific to defined embryonic brain regions. This finding indicated that a regional identity of iNPCs was not established. Upon induction, iNPCs predominantly differentiated into astrocytes. However, the differentiation potential was not fixed and could be efficiently manipulated into general or specific subtypes of neurons by expression of additional genes. Specifically, overexpression of Nurr1 and Foxa2, transcription factors specific for midbrain dopamine neuron development, drove iNPCs to yield mature midbrain dopamine neurons equipped with presynaptic DA neuronal functions. We further assessed the therapeutic potential of iNPCs in Parkinson disease model rats.
authors.affiliation: From the Graduate School of Biomedical Science and Engineering, the Hanyang Biomedical Research Institute, the Departments of Microbiology.
authors.forename: Mi-Sun
authors.initials: MS
authors.lastname: Lim
authors.affiliation: the Hanyang Biomedical Research Institute, Biochemistry and Molecular Biology, College of Medicine, Hanyang University, Seoul 133-791, Korea.
authors.forename: Mi-Yoon
authors.initials: MY
authors.lastname: Chang
authors.affiliation: the Department of Biomedical Science, Graduate School, and.
authors.forename: Sang-Mi
authors.initials: SM
authors.lastname: Kim
authors.affiliation: the Hanyang Biomedical Research Institute, Biochemistry and Molecular Biology, College of Medicine, Hanyang University, Seoul 133-791, Korea.
authors.forename: Sang-Hoon
authors.initials: SH
authors.lastname: Yi
authors.affiliation: the Department of Anatomy and Brain Disease Research Center, College of Medicine, Ajou University, Suwon 443-749, Korea, and.
authors.forename: Haeyoung
authors.initials: H
authors.lastname: Suh-Kim
authors.affiliation: the Hanyang Biomedical Research Institute, Physiology, and.
authors.forename: Sung Jun
authors.initials: SJ
authors.lastname: Jung
authors.affiliation: the Hanyang Biomedical Research Institute, Physiology, and.
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: From the Graduate School of Biomedical Science and Engineering, the Hanyang Biomedical Research Institute, Physiology, and.
authors.forename: Jin Hyuk
authors.initials: JH
authors.lastname: Kim
authors.affiliation: From the Graduate School of Biomedical Science and Engineering, the Hanyang Biomedical Research Institute, Biochemistry and Molecular Biology, College of Medicine, Hanyang University, Seoul 133-791, Korea.
authors.forename: Yong-Sung
authors.initials: YS
authors.lastname: Lee
authors.affiliation: the Departments of Microbiology.
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Lee
authors.affiliation: the Department of Physiology and Cell Therapy Center, Yonsei University College of Medicine, Seoul 120-752, Korea.
authors.forename: Dong-Wook
authors.initials: DW
authors.lastname: Kim
authors.affiliation: the Hanyang Biomedical Research Institute, Biochemistry and Molecular Biology, College of Medicine, Hanyang University, Seoul 133-791, Korea, leesh@hanyang.ac.kr.
authors.forename: Sang-Hun
authors.initials: SH
authors.lastname: Lee
authors.affiliation: From the Graduate School of Biomedical Science and Engineering, the Hanyang Biomedical Research Institute, the Departments of Microbiology, chshpark@hanyang.ac.kr.
authors.forename: Chang-Hwan
authors.initials: CH
authors.lastname: Park
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D054338:Cell Transdifferentiation; D065150:Cellular Reprogramming; D004298:Dopamine / Q000378:metabolism*; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D015870:Gene Expression; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics; D008636:Mesencephalon / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000235:genetics; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism*; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2 / Q000235:genetics; D050809:POU Domain Factors / Q000235:genetics; D020734:Parkinsonian Disorders / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D051381:Rats; D011917:Rats, Inbred Lew; D017208:Rats, Wistar; D014157:Transcription Factors / Q000235:genetics
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D004298:Dopamine; D051542:Hepatocyte Nuclear Factor 3-beta; D009419:Nerve Tissue Proteins; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D050809:POU Domain Factors; D014157:Transcription Factors; C499317:Ascl1 protein, mouse; C496503:Foxa2 protein, mouse; C561019:Myt1l protein, mouse; C539883:Nr4a2 protein, mouse; C074066:Pou3f2 protein, mouse
keywords: Parkinson disease; cell therapy; neural stem cell (NSC); neurodegeneration; reprogramming