title: Efficient generation of dopaminergic-like neurons by overexpression of Nurr1 and Pitx3 in mouse induced Pluripotent Stem Cells.
abstract: Parkinson's disease (PD) is a neurodegenerative disorder, in which the nigro-striatal Dopaminergic (DAergic) neurons are selectively lost. Treatment of neurodegenerative diseases with Pluripotent Stem Cells (PSCs) is a big interest in cell therapy. Here, we used induced Pluripotent Stem Cells (iPSCs) expressing two master Dopaminergic (DAergic) transcription factors, i.e. Nurr1 and Pitx3, to generate functional in vitro DAergic-like neurons. After establishment and characterization of Doxycycline-inducible iPSCs from mouse fibroblasts, the cells were transduced by NURR1- and PITX3-harboring lentiviruses. The Nurr1/Pitx3 -iPSCs were differentiated through a five-stage protocol to generate DAergic-like neurons. The results confirmed the efficient expression of DAergic neuron markers in the end of protocol. Beside, the generated cells could exclusively synthesize and secrete Dopamine in response to secretagogues. In conclusion, overexpression of Nurr1 and Pitx3 in iPSCs could efficiently program iPSCs into functional DAergic-like neurons. This finding may have an impact on future stem cell therapy of PD.
authors.affiliation: National Center for Transgenic Mouse Research, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Salemeh
authors.initials: S
authors.lastname: Salemi
authors.affiliation: National Center for Transgenic Mouse Research, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Parvaneh
authors.initials: P
authors.lastname: Baktash
authors.affiliation: Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Bahareh
authors.initials: B
authors.lastname: Rajaei
authors.affiliation: National Center for Transgenic Mouse Research, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Mehri
authors.initials: M
authors.lastname: Noori
authors.affiliation: Department of Pharmacology, Neuroscience Research Center, Golestan University of Medical Sciences, Gorgan, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Amini
authors.affiliation: National Center for Transgenic Mouse Research, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. Electronic address: shamsa@nigeb.ac.ir.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Shamsara
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada; Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada. Electronic address: mohammad.massumi@utoronto.ca.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Massumi
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D059290:Dopaminergic Neurons / Q000378:metabolism*; D005822:Genetic Vectors; D018398:Homeodomain Proteins / Q000378:metabolism*; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D016086:Lentivirus / Q000502:physiology; D051379:Mice; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2 / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D018398:Homeodomain Proteins; C539883:Nr4a2 protein, mouse; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D014157:Transcription Factors; C433629:homeobox protein PITX3
keywords: Dopaminergic neurons; Induced-pluripotent stem cells; Nurr1; Parkinson’s disease; Pitx3