title: In vitro characterization of neurite extension using induced pluripotent stem cells derived from lissencephaly patients with TUBA1A missense mutations.
abstract: Lissencephaly, or smooth brain, is a severe congenital brain malformation that is thought to be associated with impaired neuronal migration during corticogenesis. However, the exact etiology of lissencephaly in humans remains unknown. Research on congenital diseases is limited by the shortage of clinically derived resources, especially for rare pediatric diseases. The research on lissencephaly is further limited because gyration in humans is more evolved than that in model animals such as mice. To overcome these limitations, we generated induced pluripotent stem cells (iPSCs) from the umbilical cord and peripheral blood of two lissencephaly patients with different clinical severities carrying alpha tubulin (TUBA1A) missense mutations (Patient A, p.N329S; Patient B, p.R264C).
  
  
  
  Neural progenitor cells were generated from these iPSCs (iPSC-NPCs) using SMAD signaling inhibitors. These iPSC-NPCs expressed TUBA1A at much higher levels than undifferentiated iPSCs and, like fetal NPCs, readily differentiated into neurons. Using these lissencephaly iPSC-NPCs, we showed that the neurons derived from the iPSCs obtained from Patient A but not those obtained from Patient B showed abnormal neurite extension, which correlated with the pathological severity in the brains of the patients.
  
  
  
  We established iPSCs derived from lissencephaly patients and successfully modeled one aspect of the pathogenesis of lissencephaly in vitro using iPSC-NPCs and iPSC-derived neurons. The iPSCs from patients with brain malformation diseases helped us understand the mechanism underlying rare diseases and human corticogenesis without the use of postmortem brains.
authors.affiliation: Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan.|Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, Chuo-ku, Osaka, 540-0006, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Bamba
authors.affiliation: Division of Stem Cell Research, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Shofuda
authors.affiliation: Department of Pediatrics, Yamagata University Faculty of Medicine, 2-2-2 Iida-Nishi, Yamagata-shi, Yamagata, 990-9585, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Kato
authors.affiliation: CRIFM Clinical Research Institute of Fetal Medicine Pooh Maternity Clinic, 7-1-24 Uehommachi, Tennoji, Osaka, 543-0001, Japan.
authors.forename: Ritsuko K
authors.initials: RK
authors.lastname: Pooh
authors.affiliation: Department of Obstetrics and Gynecology, Okayama Medical Center, National Hospital Organization, 1711-1 Tamasu, Okayama, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Tateishi
authors.affiliation: Department of Pediatrics, Tokyo Women's Medical University, Yachiyo Medical Center, 477-96 Owadashinden, Yachiyo-shi, 276-8524, Japan.
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Takanashi
authors.affiliation: Department of Radiological Science, International University of Health and Welfare, Graduate School, 2-4-16, Momochihama, Sawara-ku, Fukuoka, 814-0001, Japan.
authors.forename: Hidetsuna
authors.initials: H
authors.lastname: Utsunomiya
authors.affiliation: Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, Chuo-ku, Osaka, 540-0006, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Sumida
authors.affiliation: Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, Chuo-ku, Osaka, 540-0006, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kanematsu
authors.affiliation: Laboratory Animal Research Department, Central Institute for Experimental Animals, 3-25-12 Tonomachi, Kawasaki-ku, Kawasaki, Kanagawa, 210-0821, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Suemizu
authors.affiliation: Laboratory Animal Research Department, Central Institute for Experimental Animals, 3-25-12 Tonomachi, Kawasaki-ku, Kawasaki, Kanagawa, 210-0821, Japan.
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Higuchi
authors.affiliation: Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan.
authors.forename: Wado
authors.initials: W
authors.lastname: Akamatsu
authors.affiliation: Program in Neuroscience and Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A7, Canada.|McEwen Center for Regenerative Medicine, University Health Network, Toronto, ON, M5G 0A7, Canada.|Department of Molecular Genetics, University of Toronto, Toronto, ON, M5G1X5, Canada.
authors.forename: Denis
authors.initials: D
authors.lastname: Gallagher
authors.affiliation: Program in Neuroscience and Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A7, Canada.|Department of Molecular Genetics, University of Toronto, Toronto, ON, M5G1X5, Canada.|Department of Physiology, University of Toronto, Toronto, ON, M5G 1X5, Canada.
authors.forename: Freda D
authors.initials: FD
authors.lastname: Miller
authors.affiliation: Department of Pediatric Neurosurgery, Takatsuki General Hospital, 1-3-13、Kosobe cho, Takatsuki, 569-1192, Japan.|Division of Molecular Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
authors.forename: Mami
authors.initials: M
authors.lastname: Yamasaki
authors.affiliation: Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, Chuo-ku, Osaka, 540-0006, Japan.|Department of Neurosurgery, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
authors.forename: Yonehiro
authors.initials: Y
authors.lastname: Kanemura
authors.affiliation: Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan. hidokano@a2.keio.jp.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Okano
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D002675:Child, Preschool; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000473:pathology*; D054082:Lissencephaly / Q000235:genetics*; D008279:Magnetic Resonance Imaging; D008297:Male; D020125:Mutation, Missense / Q000235:genetics*; D058953:Neural Stem Cells / Q000378:metabolism; D016501:Neurites / Q000378:metabolism*; D009457:Neuroglia / Q000378:metabolism; D051785:Smad Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism; D014404:Tubulin / Q000235:genetics*
chemical_list: D005456:Fluorescent Dyes; D051785:Smad Proteins; C533522:TUBA1A protein, human; D014404:Tubulin
keywords: Induced pluripotent stem cells; Lissencephaly; Neural progenitor cells; TUBA1A