Efficient induction of dopaminergic neuron differentiation from induced pluripotent stem cells reveals impaired mitophagy in PARK2 neurons

Sadafumi Suzuki; Wado Akamatsu; Fumihiko Kisa; Takefumi Sone; Kei ichi Ishikawa; Naoko Kuzumaki; Hiroyuki Katayama; Atsushi Miyawaki; Nobutaka Hattori; Hideyuki Okano

doi:10.1016/j.bbrc.2016.12.188

Efficient induction of dopaminergic neuron differentiation from induced pluripotent stem cells reveals impaired mitophagy in PARK2 neurons

Sadafumi Suzuki, Wado Akamatsu, Fumihiko Kisa, Takefumi Sone, Kei ichi Ishikawa, Naoko Kuzumaki, Hiroyuki Katayama, Atsushi Miyawaki, Nobutaka Hattori, Hideyuki Okano

Department of Physiology

Research output: Contribution to journal › Article › peer-review

51 Citations (Scopus)

Abstract

Patient-specific induced pluripotent stem cells (iPSCs) show promise for use as tools for in vitro modeling of Parkinson's disease. We sought to improve the efficiency of dopaminergic (DA) neuron induction from iPSCs by the using surface markers expressed in DA progenitors to increase the significance of the phenotypic analysis. By sorting for a CD184^high/CD44^- fraction during neural differentiation, we obtained a population of cells that were enriched in DA neuron precursor cells and achieved higher differentiation efficiencies than those obtained through the same protocol without sorting. This high efficiency method of DA neuronal induction enabled reliable detection of reactive oxygen species (ROS) accumulation and vulnerable phenotypes in PARK2 iPSCs-derived DA neurons. We additionally established a quantitative system using the mt-mKeima reporter system to monitor mitophagy in which mitochondria fuse with lysosomes and, by combining this system with the method of DA neuronal induction described above, determined that mitophagy is impaired in PARK2 neurons. These findings suggest that the efficiency of DA neuron induction is important for the precise detection of cellular phenotypes in modeling Parkinson's disease.

Original language	English
Pages (from-to)	88-93
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	483
Issue number	1
DOIs	https://doi.org/10.1016/j.bbrc.2016.12.188
Publication status	Published - 2017 Jan 29

Keywords

Dopaminergic neurons
Flow cytometry
Induced pluripotent stem cells
Mitophagy
Parkinson's disease

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2016.12.188

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Suzuki, S., Akamatsu, W., Kisa, F., Sone, T., Ishikawa, K. I., Kuzumaki, N., Katayama, H., Miyawaki, A., Hattori, N., & Okano, H. (2017). Efficient induction of dopaminergic neuron differentiation from induced pluripotent stem cells reveals impaired mitophagy in PARK2 neurons. Biochemical and Biophysical Research Communications, 483(1), 88-93. https://doi.org/10.1016/j.bbrc.2016.12.188

Suzuki, S, Akamatsu, W, Kisa, F, Sone, T, Ishikawa, KI, Kuzumaki, N, Katayama, H, Miyawaki, A, Hattori, N & Okano, H 2017, 'Efficient induction of dopaminergic neuron differentiation from induced pluripotent stem cells reveals impaired mitophagy in PARK2 neurons', Biochemical and Biophysical Research Communications, vol. 483, no. 1, pp. 88-93. https://doi.org/10.1016/j.bbrc.2016.12.188

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title = "Efficient induction of dopaminergic neuron differentiation from induced pluripotent stem cells reveals impaired mitophagy in PARK2 neurons",

abstract = "Patient-specific induced pluripotent stem cells (iPSCs) show promise for use as tools for in vitro modeling of Parkinson's disease. We sought to improve the efficiency of dopaminergic (DA) neuron induction from iPSCs by the using surface markers expressed in DA progenitors to increase the significance of the phenotypic analysis. By sorting for a CD184high/CD44- fraction during neural differentiation, we obtained a population of cells that were enriched in DA neuron precursor cells and achieved higher differentiation efficiencies than those obtained through the same protocol without sorting. This high efficiency method of DA neuronal induction enabled reliable detection of reactive oxygen species (ROS) accumulation and vulnerable phenotypes in PARK2 iPSCs-derived DA neurons. We additionally established a quantitative system using the mt-mKeima reporter system to monitor mitophagy in which mitochondria fuse with lysosomes and, by combining this system with the method of DA neuronal induction described above, determined that mitophagy is impaired in PARK2 neurons. These findings suggest that the efficiency of DA neuron induction is important for the precise detection of cellular phenotypes in modeling Parkinson's disease.",

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author = "Sadafumi Suzuki and Wado Akamatsu and Fumihiko Kisa and Takefumi Sone and Ishikawa, {Kei ichi} and Naoko Kuzumaki and Hiroyuki Katayama and Atsushi Miyawaki and Nobutaka Hattori and Hideyuki Okano",

note = "Funding Information: This study was supported by funding from the Project for the Realization of Regenerative Medicine and Support for Core Institutes for iPS Cell Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) to H.O., the Research Center Network for Realization Research Centers/Projects of Regenerative Medicine (the Program for Intractable Disease Research Utilizing Disease-Specific iPS Cells) from the Japan Science and Technology Agency (JST) and the Japan Agency for Medical Research and Development (AMED) to H.O., the New Energy and Industrial Technology Development Organization (NEDO) to H.O. and W.A., the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number JP16K09677 to W.A. and JP16K19524 to K.I., and a Grant in-Aid for the Global COE Program from MEXT to Keio University. H.O. is a scientific consultant for SanBio, Co. Ltd., Eisai, Co., Ltd. Publisher Copyright: {\textcopyright} 2016",

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AU - Suzuki, Sadafumi

AU - Akamatsu, Wado

AU - Kisa, Fumihiko

AU - Sone, Takefumi

AU - Ishikawa, Kei ichi

AU - Kuzumaki, Naoko

AU - Katayama, Hiroyuki

AU - Miyawaki, Atsushi

AU - Hattori, Nobutaka

AU - Okano, Hideyuki

N1 - Funding Information: This study was supported by funding from the Project for the Realization of Regenerative Medicine and Support for Core Institutes for iPS Cell Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) to H.O., the Research Center Network for Realization Research Centers/Projects of Regenerative Medicine (the Program for Intractable Disease Research Utilizing Disease-Specific iPS Cells) from the Japan Science and Technology Agency (JST) and the Japan Agency for Medical Research and Development (AMED) to H.O., the New Energy and Industrial Technology Development Organization (NEDO) to H.O. and W.A., the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number JP16K09677 to W.A. and JP16K19524 to K.I., and a Grant in-Aid for the Global COE Program from MEXT to Keio University. H.O. is a scientific consultant for SanBio, Co. Ltd., Eisai, Co., Ltd. Publisher Copyright: © 2016

PY - 2017/1/29

Y1 - 2017/1/29

N2 - Patient-specific induced pluripotent stem cells (iPSCs) show promise for use as tools for in vitro modeling of Parkinson's disease. We sought to improve the efficiency of dopaminergic (DA) neuron induction from iPSCs by the using surface markers expressed in DA progenitors to increase the significance of the phenotypic analysis. By sorting for a CD184high/CD44- fraction during neural differentiation, we obtained a population of cells that were enriched in DA neuron precursor cells and achieved higher differentiation efficiencies than those obtained through the same protocol without sorting. This high efficiency method of DA neuronal induction enabled reliable detection of reactive oxygen species (ROS) accumulation and vulnerable phenotypes in PARK2 iPSCs-derived DA neurons. We additionally established a quantitative system using the mt-mKeima reporter system to monitor mitophagy in which mitochondria fuse with lysosomes and, by combining this system with the method of DA neuronal induction described above, determined that mitophagy is impaired in PARK2 neurons. These findings suggest that the efficiency of DA neuron induction is important for the precise detection of cellular phenotypes in modeling Parkinson's disease.

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KW - Mitophagy

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Efficient induction of dopaminergic neuron differentiation from induced pluripotent stem cells reveals impaired mitophagy in PARK2 neurons

Abstract

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