In vitro characterization of neurite extension using induced pluripotent stem cells derived from lissencephaly patients with TUBA1A missense mutations View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-07-19

AUTHORS

Yohei Bamba, Tomoko Shofuda, Mitsuhiro Kato, Ritsuko K. Pooh, Yoko Tateishi, Jun-ichi Takanashi, Hidetsuna Utsunomiya, Miho Sumida, Daisuke Kanematsu, Hiroshi Suemizu, Yuichiro Higuchi, Wado Akamatsu, Denis Gallagher, Freda D. Miller, Mami Yamasaki, Yonehiro Kanemura, Hideyuki Okano

ABSTRACT

BackgroundLissencephaly, 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).ResultsNeural 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.ConclusionWe 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. More... »

PAGES

70

Journal

TITLE

Molecular Brain

ISSUE

1

VOLUME

9

Author Affiliations

  • Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, 540-0006, Chuo-ku, Osaka, Japan
  • Division of Stem Cell Research, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan
  • Department of Pediatrics, Yamagata University Faculty of Medicine, 2-2-2 Iida-Nishi, 990-9585, Yamagata-shi, Yamagata, Japan
  • CRIFM Clinical Research Institute of Fetal Medicine Pooh Maternity Clinic, 7-1-24 Uehommachi, 543-0001, Tennoji, Osaka, Japan
  • Department of Obstetrics and Gynecology, Okayama Medical Center, National Hospital Organization, 1711-1 Tamasu, Okayama, Japan
  • Department of Pediatrics, Tokyo Women’s Medical University, Yachiyo Medical Center, 477-96 Owadashinden, 276-8524, Yachiyo-shi, Japan
  • Department of Radiological Science, International University of Health and Welfare, Graduate School, 2-4-16, Momochihama, 814-0001, Sawara-ku, Fukuoka, Japan
  • Laboratory Animal Research Department, Central Institute for Experimental Animals, 3-25-12 Tonomachi, 210-0821, Kawasaki-ku, Kawasaki, Kanagawa, Japan
  • Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, 160-8582, Shinjuku-ku, Tokyo, Japan
  • Department of Molecular Genetics, University of Toronto, M5G1X5, Toronto, ON, Canada
  • Department of Physiology, University of Toronto, M5G 1X5, Toronto, ON, Canada
  • Division of Molecular Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan
  • Department of Neurosurgery, Osaka National Hospital, National Hospital Organization, Osaka, Japan
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/s13041-016-0246-y

    DOI

    http://dx.doi.org/10.1186/s13041-016-0246-y

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1029466309

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/27431206


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        "description": "BackgroundLissencephaly, 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).ResultsNeural 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.ConclusionWe 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.", 
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    351 grid-institutes:grid.416803.8 schema:alternateName Department of Neurosurgery, Osaka National Hospital, National Hospital Organization, Osaka, Japan
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    353 Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, 540-0006, Chuo-ku, Osaka, Japan
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    359 Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, 540-0006, Chuo-ku, Osaka, Japan
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