title: Generation of Naivetropic Induced Pluripotent Stem Cells from Parkinson's Disease Patients for High-Efficiency Genetic Manipulation and Disease Modeling.
abstract: The lack of robust Parkinson's disease (PD) phenotype in parkin knockout rodents and the identification of defective dopaminergic (DA) neurotransmission in midbrain DA neurons derived from induced pluripotent stem cells (iPSC) of PD patients with parkin mutations demonstrate the utility of patient-specific iPSCs as an effective system to model the unique vulnerabilities of midbrain DA neurons in PD. Significant efforts have been directed at developing efficient genomic engineering technologies in human iPSCs to study diseases such as PD. In the present study, we converted patient-specific iPSCs from the primed state to a naivetropic state by DOX-induced expression of transgenes (Oct4, Sox2, Klf4, c-Myc, and Nanog) and the use of 2iL (MEK inhibitor PD0325901, GSK3 inhibitor CHIR99021, and human LIF). These patient-specific naivetropic iPSCs were pluripotent in terms of marker expression, spontaneous differentiation in vitro, and teratoma formation in vivo. They exhibited morphological, proliferative, and clonogenic characteristics very similar to naive mouse embryonic stem cells (ESC). The high clonal efficiency and proliferation rate of naivetropic iPSCs enabled very efficient gene targeting of GFP to the PITX3 locus by transcription activator-like effector nuclease. The naivetropic iPSCs could be readily reverted to the primed state upon the withdrawal of DOX, 2iL, and the switch to primed-state hESC culture conditions. Midbrain DA neurons differentiated from the reverted iPSCs retained the original phenotypes caused by parkin mutations, attesting to the robustness of these phenotypes and the usefulness of genomic engineering in patient-specific naivetropic iPSCs for studying PD.
authors.affiliation: 1 Department of Physiology and Biophysics, State University of New York at Buffalo , Buffalo, New York.|2 Veterans Affairs Western New York Healthcare System , Buffalo, New York.
authors.forename: Zhixing
authors.initials: Z
authors.lastname: Hu
authors.affiliation: 1 Department of Physiology and Biophysics, State University of New York at Buffalo , Buffalo, New York.|3 Department of Neurology, Second Affiliated Hospital, College of Medicine, Zhejiang University , Hangzhou, China .
authors.forename: Jiali
authors.initials: J
authors.lastname: Pu
authors.affiliation: 1 Department of Physiology and Biophysics, State University of New York at Buffalo , Buffalo, New York.|2 Veterans Affairs Western New York Healthcare System , Buffalo, New York.
authors.forename: Houbo
authors.initials: H
authors.lastname: Jiang
authors.affiliation: 1 Department of Physiology and Biophysics, State University of New York at Buffalo , Buffalo, New York.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhong
authors.affiliation: 4 Department of Pathology and Laboratory Medicine, Roswell Park Cancer Institute , Buffalo, New York.
authors.forename: Jingxin
authors.initials: J
authors.lastname: Qiu
authors.affiliation: 5 Department of Neurobiology, Key Laboratory for Neurodegenerative Disorders of the Ministry of Education, Beijing Institute for Brain Disorders, Capital Medical University , Beijing, China .
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.affiliation: 5 Department of Neurobiology, Key Laboratory for Neurodegenerative Disorders of the Ministry of Education, Beijing Institute for Brain Disorders, Capital Medical University , Beijing, China .
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Wang
authors.affiliation: 3 Department of Neurology, Second Affiliated Hospital, College of Medicine, Zhejiang University , Hangzhou, China .
authors.forename: Baorong
authors.initials: B
authors.lastname: Zhang
authors.affiliation: 1 Department of Physiology and Biophysics, State University of New York at Buffalo , Buffalo, New York.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yan
authors.affiliation: 1 Department of Physiology and Biophysics, State University of New York at Buffalo , Buffalo, New York.|2 Veterans Affairs Western New York Healthcare System , Buffalo, New York.|5 Department of Neurobiology, Key Laboratory for Neurodegenerative Disorders of the Ministry of Education, Beijing Institute for Brain Disorders, Capital Medical University , Beijing, China .
authors.forename: Jian
authors.initials: J
authors.lastname: Feng
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D059290:Dopaminergic Neurons / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D000090062:Kruppel-Like Factor 4; D051379:Mice; D010300:Parkinson Disease / Q000235:genetics*; D039904:Pluripotent Stem Cells / Q000166:cytology*