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Mol Brain. 2016 Jul 19;9(1):70. doi: 10.1186/s13041-016-0246-y.

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

Author information

1
Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan.
2
Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, 2-1-14 Hoenzaka, Chuo-ku, Osaka, 540-0006, Japan.
3
Division of Stem Cell Research, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
4
Department of Pediatrics, Yamagata University Faculty of Medicine, 2-2-2 Iida-Nishi, Yamagata-shi, Yamagata, 990-9585, Japan.
5
CRIFM Clinical Research Institute of Fetal Medicine Pooh Maternity Clinic, 7-1-24 Uehommachi, Tennoji, Osaka, 543-0001, Japan.
6
Department of Obstetrics and Gynecology, Okayama Medical Center, National Hospital Organization, 1711-1 Tamasu, Okayama, Japan.
7
Department of Pediatrics, Tokyo Women's Medical University, Yachiyo Medical Center, 477-96 Owadashinden, Yachiyo-shi, 276-8524, Japan.
8
Department of Radiological Science, International University of Health and Welfare, Graduate School, 2-4-16, Momochihama, Sawara-ku, Fukuoka, 814-0001, Japan.
9
Laboratory Animal Research Department, Central Institute for Experimental Animals, 3-25-12 Tonomachi, Kawasaki-ku, Kawasaki, Kanagawa, 210-0821, Japan.
10
Program in Neuroscience and Mental Health, The Hospital for Sick Children, Toronto, ON, M5G 0A7, Canada.
11
McEwen Center for Regenerative Medicine, University Health Network, Toronto, ON, M5G 0A7, Canada.
12
Department of Molecular Genetics, University of Toronto, Toronto, ON, M5G1X5, Canada.
13
Department of Physiology, University of Toronto, Toronto, ON, M5G 1X5, Canada.
14
Department of Pediatric Neurosurgery, Takatsuki General Hospital, 1-3-13、Kosobe cho, Takatsuki, 569-1192, Japan.
15
Division of Molecular Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
16
Department of Neurosurgery, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
17
Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan. hidokano@a2.keio.jp.

Abstract

BACKGROUND:

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).

RESULTS:

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.

CONCLUSION:

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.

KEYWORDS:

Induced pluripotent stem cells; Lissencephaly; Neural progenitor cells; TUBA1A

PMID:
27431206
PMCID:
PMC4950778
DOI:
10.1186/s13041-016-0246-y
[Indexed for MEDLINE]
Free PMC Article

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