title: Homogeneous generation of iDA neurons with high similarity to bona fide DA neurons using a drug inducible system.
abstract: Recent work generating induced dopaminergic (iDA) neurons using direct lineage reprogramming potentially provides a novel platform for the study and treatment Parkinson's disease (PD). However, one of the most important issues for iDA-based applications is the degree to which iDA neurons resemble the molecular and functional properties of their endogenous DA neuron counterparts. Here we report that the homogeneity of the reprogramming gene expression system is critical for the generation of iDA neuron cultures that are highly similar to endogenous DA neurons. We employed an inducible system that carries iDA-inducing factors as defined transgenes for direct lineage reprogramming to iDA neurons. This system circumvents the need for viral transduction, enabling a more efficient and reproducible reprogramming process for the generation of genetically homogenous iDA neurons. We showed that this inducible system generates iDA neurons with high similarity to their bona fide in vivo counterparts in comparison to direct infection methods. Thus, our results suggest that homogenous expression of exogenous genes in direct lineage reprogramming is critical for the generation of high quality iDA neuron cultures, making such culture systems a valuable resource for iDA-based drug screening and, ultimately, potential therapeutic intervention in PD.
authors.affiliation: Lab of Stem Cells & Cell Reprogramming, Department of Biomedical Engineering, Dongguk University, Seoul, 100-715, Republic of Korea.
authors.forename: Hanseul
authors.initials: H
authors.lastname: Park
authors.affiliation: Lab of Stem Cells & Cell Reprogramming, Department of Biomedical Engineering, Dongguk University, Seoul, 100-715, Republic of Korea.
authors.forename: Hongwon
authors.initials: H
authors.lastname: Kim
authors.affiliation: Lab of Stem Cells & Cell Reprogramming, Department of Biomedical Engineering, Dongguk University, Seoul, 100-715, Republic of Korea.
authors.forename: Junsang
authors.initials: J
authors.lastname: Yoo
authors.affiliation: Center for Neural Science and Functional Connectomics, Brain Science Institute, Korea Institute of Science and Technology, Seoul, 136-791, Republic of Korea.
authors.forename: Jaekwang
authors.initials: J
authors.lastname: Lee
authors.affiliation: Lab of Stem Cells & Cell Reprogramming, Department of Biomedical Engineering, Dongguk University, Seoul, 100-715, Republic of Korea.
authors.forename: Hwan
authors.initials: H
authors.lastname: Choi
authors.affiliation: Lab of Stem Cells & Cell Reprogramming, Department of Biomedical Engineering, Dongguk University, Seoul, 100-715, Republic of Korea.
authors.forename: Soonbong
authors.initials: S
authors.lastname: Baek
authors.affiliation: Center for Neural Science and Functional Connectomics, Brain Science Institute, Korea Institute of Science and Technology, Seoul, 136-791, Republic of Korea.
authors.forename: C Justin
authors.initials: CJ
authors.lastname: Lee
authors.affiliation: Stem Cell Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBBB), Korea University of Science & Technology (UST), Daejeon 305-333, Republic of Korea.
authors.forename: Janghwan
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Lengner
authors.affiliation: Department of Life Science, Dongguk University, Seoul, 100-715, Republic of Korea.
authors.forename: Jung-Suk
authors.initials: JS
authors.lastname: Sung
authors.affiliation: Lab of Stem Cells & Cell Reprogramming, Department of Biomedical Engineering, Dongguk University, Seoul, 100-715, Republic of Korea. Electronic address: jk2316@gmail.com.
authors.forename: Jongpil
authors.initials: J
authors.lastname: Kim
mesh_terms: D000818:Animals; D019070:Cell Lineage / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D065150:Cellular Reprogramming / Q000187:drug effects; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D004318:Doxycycline / Q000494:pharmacology*; D055724:Electrophysiological Phenomena / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D018398:Homeodomain Proteins / Q000378:metabolism; D051379:Mice; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D018398:Homeodomain Proteins; D014157:Transcription Factors; C485184:enhanced green fluorescent protein; C433629:homeobox protein PITX3; D049452:Green Fluorescent Proteins; D004318:Doxycycline
keywords: Direct lineage reprogramming; Gene expression system; Induced neurons