Advertisement
No access
Report
AMYOTROPHIC LATERAL SCLEROSIS

The Src/c-Abl pathway is a potential therapeutic target in amyotrophic lateral sclerosis

Science Translational Medicine
24 May 2017
Vol 9, Issue 391

A stepping stone to ALS drug discovery

ALS is a heterogeneous motor neuron disease for which there is no treatment and for which a common therapeutic target has yet to be identified. In a new study, Imamura et al. developed a drug screen using motor neurons generated from ALS patient induced pluripotent stem cells (iPSCs). They screened existing drugs and showed that inhibitors of Src/c-Abl kinases promoted autophagy and rescued ALS motor neurons from degeneration. One of the drugs was effective for promoting survival of motor neurons derived from ALS patients with different genetic mutations. The Src/c-Abl pathway may be a potential therapeutic target for developing new drugs to treat ALS.

Abstract

Amyotrophic lateral sclerosis (ALS), a fatal disease causing progressive loss of motor neurons, still has no effective treatment. We developed a phenotypic screen to repurpose existing drugs using ALS motor neuron survival as readout. Motor neurons were generated from induced pluripotent stem cells (iPSCs) derived from an ALS patient with a mutation in superoxide dismutase 1 (SOD1). Results of the screen showed that more than half of the hits targeted the Src/c-Abl signaling pathway. Src/c-Abl inhibitors increased survival of ALS iPSC-derived motor neurons in vitro. Knockdown of Src or c-Abl with small interfering RNAs (siRNAs) also rescued ALS motor neuron degeneration. One of the hits, bosutinib, boosted autophagy, reduced the amount of misfolded mutant SOD1 protein, and attenuated altered expression of mitochondrial genes. Bosutinib also increased survival in vitro of ALS iPSC-derived motor neurons from patients with sporadic ALS or other forms of familial ALS caused by mutations in TAR DNA binding protein (TDP-43) or repeat expansions in C9orf72. Furthermore, bosutinib treatment modestly extended survival of a mouse model of ALS with an SOD1 mutation, suggesting that Src/c-Abl may be a potentially useful target for developing new drugs to treat ALS.

Get full access to this article

View all available purchase options and get full access to this article.

Supplementary Material

Summary

Materials and methods
Fig. S1. Generation of iPSCs from control and ALS patients.
Fig. S2. Characterization of motor neurons and genetic correction of mutant SOD1 iPSCs.
Fig. S3. Investigation of the effects of Src/c-Abl inhibitors.
Fig. S4. mRNA expression changes after bosutinib treatment by single-cell analysis.
Fig. S5. Decrease in misfolded proteins after bosutinib treatment.
Fig. S6. Analysis of postmortem ALS spinal cord tissue.
Table S1. List of iPSC clones.
Table S2. Sequence variations in exon regions for sporadic ALS.
Table S3. List of hit compounds.
Table S4. Genes highly expressed in mutant SOD1 ALS motor neurons identified by single-cell RNA-seq.
Table S5. Genes highly expressed in control motor neurons identified by single-cell RNA-seq.
Table S6. List of postmortem spinal cord tissue for immunohistochemistry.
Table S7. List of postmortem spinal cord tissue for ELISA.
Table S8. Primer list for editing of SOD1 gene.
Table S9. Primer list for quantitative PCR.
References (4856)

Resources

File (aaf3962_sm.pdf)

REFERENCES AND NOTES

1
S.-C. Ling, M. Polymenidou, D. W. Cleveland, Converging mechanisms in ALS and FTD: Disrupted RNA and protein homeostasis. Neuron 79, 416–438 (2013).
2
J. Ravits, S. Appel, R. H. Baloh, R. Barohn, B. R. Brooks, L. Elman, M. K. Floeter, C. Henderson, C. Lomen-Hoerth, J. D. Macklis, L. McCluskey, H. Mitsumoto, S. Przedborski, J. Rothstein, J. Q. Trojanowski, L. H. van den Berg, S. Ringel, Deciphering amyotrophic lateral sclerosis: What phenotype, neuropathology and genetics are telling us about pathogenesis. Amyotrophic. Lateral Scler. Frontotemporal Degener. 14 (suppl. 1), 5–18 (2013).
3
D. R. Rosen, T. Siddique, D. Patterson, D. A. Figlewicz, P. Sapp, A. Hentati, D. Donaldson, J. Goto, J. P. O’Regan, H.-X. Deng, Z. Rahmani, A. Krizus, D. Mckenna-Yasek, A. Cayabyab, S. M. Gaston, R. Berger, R. E. Tanzi, J. J. Halperin, B. Herzfeldt, R. van den Bergh, W.-Y. Hung, T. Bird, G. Deng, D. W. Mulder, C. Smyth, N. G. Laing, E. Soriano, M. A. Pericak-Vance, J. Haines, G. A. Rouleau, J. S. Gusella, H. R. Horvitz, R. H. Brown Jr, Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature 362, 59–62 (1993).
4
M. E. Gurney, H. Pu, A. Y. Chiu, M. C. Dal Canto, C. Y. Polchow, D. D. Alexander, J. Caliendo, A. Hentati, Y. W. Kwon, H.-X. Deng, W. Chen, P. Zhai, R. L. Sufit, T. Siddique, Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation. Science 264, 1772–1775 (1994).
5
N. Egawa, S. Kitaoka, K. Tsukita, M. Naitoh, K. Takahashi, T. Yamamoto, F. Adachi, T. Kondo, K. Okita, I. Asaka, T. Aoi, A. Watanabe, Y. Yamada, A. Morizane, J. Takahashi, T. Ayaki, H. Ito, K. Yoshikawa, S. Yamawaki, S. Suzuki, D. Watanabe, H. Hioki, T. Kaneko, K. Makioka, K. Okamoto, H. Takuma, A. Tamaoka, K. Hasegawa, T. Nonaka, M. Hasegawa, A. Kawata, M. Yoshida, T. Nakahata, R. Takahashi, M. C. N. Marchetto, F. H. Gage, S. Yamanaka, H. Inoue, Drug screening for ALS using patient-specific induced pluripotent stem cells. Sci. Transl. Med. 4, 145ra104 (2012).
6
B. Bilican, A. Serio, S. J. Barmada, A. L. Nishimura, G. J. Sullivan, M. Carrasco, H. P. Phatnani, C. A. Puddifoot, D. Story, J. Fletcher, I.-H. Park, B. A. Friedman, G. Q. Daley, D. J. A. Wyllie, G. E. Hardingham, I. Wilmut, S. Finkbeiner, T. Maniatis, C. E. Shaw, S. Chandran, Mutant induced pluripotent stem cell lines recapitulate aspects of TDP-43 proteinopathies and reveal cell-specific vulnerability. Proc. Natl. Acad. Sci. U.S.A. 109, 5803–5808 (2012).
7
E. Kiskinis, J. Sandoe, L. A. Williams, G. L. Boulting, R. Moccia, B. J. Wainger, S. Han, T. Peng, S. Thams, S. Mikkilineni, C. Mellin, F. T. Merkle, B. N. Davis-Dusenbery, M. Ziller, D. Oakley, J. Ichida, S. Di Costanzo, N. Atwater, M. L. Maeder, M. J. Goodwin, J. Nemesh, R. E. Handsaker, D. Paull, S. Noggle, S. A. McCarroll, J. K. Joung, C. J. Woolf, R. H. Brown, K. Eggan, Pathways disrupted in human ALS motor neurons identified through genetic correction of mutant SOD1. Cell Stem Cell 14, 781–795 (2014).
8
M. F. Burkhardt, F. J. Martinez, S. Wright, C. Ramos, D. Volfson, M. Mason, J. Garnes, V. Dang, J. Lievers, U. Shoukat-Mumtaz, R. Martinez, H. Gai, R. Blake, E. Vaisberg, M. Grskovic, C. Johnson, S. Irion, J. Bright, B. Cooper, L. Nguyen, I. Griswold-Prenner, A. Javaherian, A cellular model for sporadic ALS using patient-derived induced pluripotent stem cells. Mol. Cell. Neurosci. 56, 355–364 (2013).
9
H. Chen, K. Qian, Z. Du, J. Cao, A. Petersen, H. Liu, L. W. Blackbourn IV, C.-L. Huang, A. Errigo, Y. Yin, J. Lu, M. Ayala, S.-C. Zhang, Modeling ALS with iPSCs reveals that mutant SOD1 misregulates neurofilament balance in motor neurons. Cell Stem Cell 14, 796–809 (2014).
10
C. J. Donnelly, P.-W. Zhang, J. T. Pham, A. R. Haeusler, N. A. Mistry, S. Vidensky, E. L. Daley, E. M. Poth, B. Hoover, D. M. Fines, N. Maragakis, P. J. Tienari, L. Petrucelli, B. J. Traynor, J. Wang, F. Rigo, C. F. Bennett, S. Blackshaw, R. Sattler, J. D. Rothstein, RNA toxicity from the ALS/FTD C9ORF72 expansion is mitigated by antisense intervention. Neuron 80, 415–428 (2013).
11
M. Mitne-Neto, M. Machado-Costa, M. C. N. Marchetto, M. H. Bengtson, C. A. Joazeiro, H. Tsuda, H. J. Bellen, H. C. A. Silva, A. S. B. Oliveira, M. Lazar, A. R. Muotri, M. Zatz, Downregulation of VAPB expression in motor neurons derived from induced pluripotent stem cells of ALS8 patients. Hum. Mol. Genet. 20, 3642–3652 (2011).
12
K. Woltjen, I. P. Michael, P. Mohseni, R. Desai, M. Mileikovsky, R. Hämäläinen, R. Cowling, W. Wang, P. Liu, M. Gertsenstein, K. Kaji, H.-K. Sung, A. Nagy, piggyBac transposition reprograms fibroblasts to induced pluripotent stem cells. Nature 458, 766–770 (2009).
13
T. T. Ashburn, K. B. Thor Drug repositioning: Identifying and developing new uses for existing drugs. Nat. Rev. Drug Discov. 3, 673–683 (2004).
14
J. S. Brugge, R. L. Erikson, Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature 269, 346–348 (1977).
15
E. K. Greuber, P. Smith-Pearson, J. Wang, A. M. Pendergast, Role of ABL family kinases in cancer: From leukaemia to solid tumours. Nat. Rev. Cancer 13, 559–571 (2013).
16
P. J. Welch, J. Y. J. Wang, A C-terminal protein-binding domain in the retinoblastoma protein regulates nuclear c-Abl tyrosine kinase in the cell cycle. Cell 75, 779–790 (1993).
17
Y. Huang, Z.-M. Yuan, T. Ishiko, S. Nakada, T. Utsugisawa, T. Kato, S. Kharbanda, D. W. Kufe, Pro-apoptotic effect of the c-Abl tyrosine kinase in the cellular response to 1-β-D-arabinofuranosylcytosine. Oncogene 15, 1947–1952 (1997).
18
J. Mojsilovic-Petrovic, G.-B. Jeong, A. Crocker, A. Arneja, S. David, D. Russell, R. G. Kalb, Protecting motor neurons from toxic insult by antagonism of adenosine A2a and Trk receptors. J. Neurosci. 26, 9250–9263 (2006).
19
C. E. Ellis, P. L. Schwartzberg, T. L. Grider, D. W. Fink, R. L. Nussbaum, α-Synuclein is phosphorylated by members of the Src family of protein-tyrosine kinases. J. Biol. Chem. 276, 3879–3884 (2001).
20
S. D. Schlatterer, C. M. Acker, P. Davies, c-Abl in neurodegenerative disease. J. Mol. Neurosci. 45, 445–452 (2011).
21
T. M. Dawson, V. L. Dawson, Parkin plays a role in sporadic Parkinson's disease. Neurodegener Dis 13, 69–71 (2014).
22
Z. Jing, J. Caltagarone, R. Bowser, Altered subcellular distribution of c-Abl in Alzheimer's disease. J. Alzheimers Dis. 17, 409–422 (2009).
23
M. A. Tremblay, C. M. Acker, P. Davies, Tau phosphorylated at tyrosine 394 is found in Alzheimer's disease tangles and can be a product of the Abl-related kinase, Arg. J. Alzheimer’s Dis. 19, 721–733 (2010).
24
R. Katsumata, S. Ishigaki, M. Katsuno, K. Kawai, J. Sone, Z. Huang, H. Adachi, F. Tanaka, F. Urano, G. Sobue, c-Abl inhibition delays motor neuron degeneration in the G93A mouse, an animal model of amyotrophic lateral sclerosis. PLOS ONE 7, e46185 (2012).
25
M. E. Hester, M. J. Murtha, S. Song, M. Rao, C. J. Miranda, K. Meyer, J. Tian, G. Boulting, D. V. Schaffer, M. X. Zhu, S. L. Pfaff, F. H. Gage, B. K. Kaspar, Rapid and efficient generation of functional motor neurons from human pluripotent stem cells using gene delivered transcription factor codes. Mol. Ther. 19, 1905–1912 (2011).
26
T. Fujisawa, K. Homma, N. Yamaguchi, H. Kadowaki, N. Tsuburaya, I. Naguro, A. Matsuzawa, K. Takeda, Y. Takahashi, J. Goto, S. Tsuji, H. Nishitoh, H. Ichijo, A novel monoclonal antibody reveals a conformational alteration shared by amyotrophic lateral sclerosis-linked SOD1 mutants. Ann. Neurol. 72, 739–749 (2012).
27
F. Gros-Louis, G. Soucy, R. Lariviere, J.-P. Julien, Intracerebroventricular infusion of monoclonal antibody or its derived Fab fragment against misfolded forms of SOD1 mutant delays mortality in a mouse model of ALS. J. Neurochem. 113, 1188–1199 (2010).
28
Y. M. Yang, S. K. Gupta, K. J. Kim, B. E. Powers, A. Cerqueira, B. J. Wainger, H. D. Ngo, K. A. Rosowski, P. A. Schein, C. A. Ackeifi, A. C. Arvanites, L. S. Davidow, C. J. Woolf, L. L. Rubin, A small molecule screen in stem-cell-derived motor neurons identifies a kinase inhibitor as a candidate therapeutic for ALS. Cell Stem Cell 12, 713–726 (2013).
29
T. E. Brotherton, Y. Li, D. Cooper, M. Gearing, J.-P. Julien, J. D. Rothstein, K. Boylan, J. D. Glass, Localization of a toxic form of superoxide dismutase 1 protein to pathologically affected tissues in familial ALS. Proc. Natl. Acad. Sci. U.S.A. 109, 5505–5510 (2012).
30
A.-L. Mahul-Mellier, B. Fauvet, A. Gysbers, I. Dikiy, A. Oueslati, S. Georgeon, A. J. Lamontanara, A. Bisquertt, D. Eliezer, E. Masliah, G. Halliday, O. Hantschel, H. A. Lashuel, c-Abl phosphorylates α-synuclein and regulates its degradation: Implication for α-synuclein clearance and contribution to the pathogenesis of Parkinson's disease. Hum. Mol. Genet. 23, 2858–2879 (2014).
31
A. Ertmer, V. Huber, S. Gilch, T. Yoshimori, V. Erfle, J. Duyster, H.-P. Elsässer, H. M. Schatzl, The anticancer drug imatinib induces cellular autophagy. Leukemia 21, 936–942 (2007).
32
I. Choi, H. D. Song, S. Lee, Y. I. Yang, J. H. Nam, S. J. Kim, J.-J. Sung, T. Kang, J. Yi, Direct observation of defects and increased ion permeability of a membrane induced by structurally disordered Cu/Zn-superoxide dismutase aggregates. PLOS ONE 6, e28982 (2011).
33
B. J. Wainger, E. Kiskinis, C. Mellin, O. Wiskow, S. S. W. Han, J. Sandoe, N. P. Perez, L. A. Williams, S. Lee, G. Boulting, J. D. Berry, R. H. Brown Jr, M. E. Cudkowicz, B. P. Bean, K. Eggan, C. J. Woolf, Intrinsic membrane hyperexcitability of amyotrophic lateral sclerosis patient-derived motor neurons. Cell Rep. 7, 1–11 (2014).
34
A.-C. Devlin, K. Burr, S. Borooah, J. D. Foster, E. M. Cleary, I. Geti, L. Vallier, C. E. Shaw, S. Chandran, G. B. Miles, Human iPSC-derived motoneurons harbouring TARDBP or C9ORF72 ALS mutations are dysfunctional despite maintaining viability. Nat. Commun. 6, 5999 (2015).
35
G. Le Masson, S. Przedborski, L. F. Abbott, A computational model of motor neuron degeneration. Neuron 83, 975–988 (2014).
36
F. Musumeci, S. Schenone, C. Brullo, M. Botta, An update on dual Src/Abl inhibitors. Future Med. Chem. 4, 799–822 (2012).
37
M. Soda, Y. L. Choi, M. Enomoto, S. Takada, Y. Yamashita, S. Ishikawa, S.-i. Fujiwara, H. Watanabe, K. Kurashina, H. Hatanaka, M. Bando, S. Ohno, Y. Ishikawa, H. Aburatani, T. Niki, Y. Sohara, Y. Sugiyama, H. Mano, Identification of the transforming EML4-ALK fusion gene in non-small-cell lung cancer. Nature 448, 561–566 (2007).
38
Y. Ito, P. Pandey, N. Mishra, S. Kumar, N. Narula, S. Kharbanda, S. Saxena, D. Kufe, Targeting of the c-Abl tyrosine kinase to mitochondria in endoplasmic reticulum stress-induced apoptosis. Mol. Cell. Biol. 21, 6233–6242 (2001).
39
Y.-S. Fang, K.-J. Tsai, Y.-J. Chang, P. Kao, R. Woods, P.-H. Kuo, C.-C. Wu, J.-Y. Liao, S.-C. Chou, V. Lin, L.-W. Jin, H. S. Yuan, I. H. Cheng, P.-H. Tu, Y.-R. Chen, Full-length TDP-43 forms toxic amyloid oligomers that are present in frontotemporal lobar dementia-TDP patients. Nat. Commun. 5, 4824 (2014).
40
M. Neumann, D. M. Sampathu, L. K. Kwong, A. C. Truax, M. C. Micsenyi, T. T. Chou, J. Bruce, T. Schuck, M. Grossman, C. M. Clark, L. F. McCluskey, B. L. Miller, E. Masliah, I. R. Mackenzie, H. Feldman, W. Feiden, H. A. Kretzschmar, J. Q. Trojanowski, V. M.-Y. Lee, Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Science 314,130–133 (2006).
41
C. Wenqiang, I. Lonskaya, M. L. Hebron, Z. Ibrahim, R. T. Olszewski, J. H. Neale, C. E.-H. Moussa, Parkin-mediated reduction of nuclear and soluble TDP-43 reverses behavioral decline in symptomatic mice. Hum. Mol. Genet. 23, 4960–4969 (2014).
42
K. J. Martin, J. S. C. Arthur, Selective kinase inhibitors as tools for neuroscience research. Neuropharmacology 63, 1227–1237 (2012).
43
T. J. Boggon, M. J. Eck, Structure and regulation of Src family kinases. Oncogene 23, 7918–7927 (2004).
44
J. D. Rothstein, Of mice and men: Reconciling preclinical ALS mouse studies and human clinical trials. Ann. Neurol. 53, 423–426 (2003).
45
K. Takahashi, K. Tanabe, M. Ohnuki, M. Narita, T. Ichisaka, K. Tomoda, S. Yamanaka, Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872 (2007).
46
K. Okita, T. Yamakawa, Y. Matsumura, Y. Sato, N. Amano, A. Watanabe, N. Goshima, S. Yamanaka, An efficient nonviral method to generate integration-free human-induced pluripotent stem cells from cord blood and peripheral blood cells. Stem Cells 31, 458–466 (2013).
47
N. Fusaki, H. Ban, A. Nishiyama, K. Saeki, M. Hasegawa, Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome. Proc. Jpn. Acad. Ser. B Phys. Biol. Sci. 85, 348–362 (2009).
48
H. L. Li, N. Fujimoto, N. Sasakawa, S. Shirai, T. Ohkame, T. Sakuma, M. Tanaka, N. Amano, A. Watanabe, H. Sakurai, T. Yamamoto, S. Yamanaka, A. Hotta, Precise correction of the dystrophin gene in duchenne muscular dystrophy patient induced pluripotent stem cells by TALEN and CRISPR-Cas9. Stem Cell Rep. 4, 143–154 (2015).
49
H. Matsui, N. Fujimoto, N. Sasakawa, Y. Ohinata, M. Shima, S. Yamanaka, M. Sugimoto, A. Hotta, Delivery of full-length factor VIII using a piggyBac transposon vector to correct a mouse model of hemophilia A. PLOS ONE 9, e104957 (2014).
50
S.-I. Kim, F. Oceguera-Yanez, C. Sakurai, M. Nakagawa, S. Yamanaka, K. Woltjen, Inducible transgene expression in human iPS cells using versatile all-in-one piggyBac transposons. Methods Mol. Biol. 1357, 111–131 (2015).
51
G. B. Miles, D. C. Yohn, H. Wichterle, T. M. Jessell, V. F. Rafuse, R. M. Brownstone, Functional properties of motoneurons derived from mouse embryonic stem cells. J. Neurosci. 24, 7848–7858 (2004).
52
T. Kondo, M. Asai, K. Tsukita, Y. Kutoku, Y. Ohsawa, Y. Sunada, K. Imamura, N. Egawa, N. Yahata, K. Okita, K. Takahashi, I. Asaka, T. Aoi, A. Watanabe, K. Watanabe, C. Kadoya, R. Nakano, D. Watanabe, K. Maruyama, O. Hori, S. Hibino, T. Choshi, T. Nakahata, H. Hioki, T. Kaneko, M. Naitoh, K. Yoshikawa, S. Yamawaki, S. Suzuki, R. Hata, S. Ueno, T. Seki, K. Kobayashi, T. Toda, K. Murakami, K. Irie, W. L. Klein, H. Mori, T. Asada, R. Takahashi, N. Iwata, S. Yamanaka, H. Inoue, Modeling Alzheimer's disease with iPSCs reveals stress phenotypes associated with intracellular Aβ and differential drug responsiveness. Cell Stem Cell 12, 487–496 (2013).
53
A. Van Hoecke, L. Schoonaert, R. Lemmens, M. Timmers, K. A. Staats, A. S. Laird, E. Peeters, T. Philips, A. Goris, B. Dubois, P. M. Andersen, A. Al-Chalabi, V. Thijs, A. M. Turnley, P. W. van Vught, J. H. Veldink, O. Hardiman, L. Van Den Bosch, P. Gonzalez-Perez, P. Van Damme, R. H. Brown Jr, L. H. van den Berg, W. Robberecht, EPHA4 is a disease modifier of amyotrophic lateral sclerosis in animal models and in humans. Nat. Med. 18, 1418–1422 (2012).
54
C. S. Lobsiger, S. Boillee, C. Pozniak, A. M. Khan, M. McAlonis-Downes, J. W. Lewcock, D. W. Cleveland, C1q induction and global complement pathway activation do not contribute to ALS toxicity in mutant SOD1 mice. Proc. Natl. Acad. Sci. U.S.A. 110, E4385–E4392 (2013).
55
T. Fujisawa, M. Takahashi, Y. Tsukamoto, N. Yamaguchi, M. Nakoji, M. Endo, H. Kodaira, Y. Hayashi, H. Nishitoh, I. Naguro, K. Homma, H. Ichijo, The ASK1-specific inhibitors K811 and K812 prolong survival in a mouse model of amyotrophic lateral sclerosis. Hum. Mol. Genet. 25, 245–253 (2016).
56
B. R. Brooks, R. G. Miller, M. Swash, T. L. Munsat, El Escorial revisited: Revised criteria for the diagnosis of amyotrophic lateral sclerosis. Amyotroph. Lateral Scler. Other Motor Neuron Disord. 1, 293–299 (2000).

(0)eLetters

eLetters is an online forum for ongoing peer review. Submission of eLetters are open to all. eLetters are not edited, proofread, or indexed. Please read our Terms of Service before submitting your own eLetter.

Log In to Submit a Response

No eLetters have been published for this article yet.

Information & Authors

Information

Published In

Science Translational Medicine
Volume 9 | Issue 391
May 2017

Submission history

Received: 15 April 2015
Accepted: 13 December 2016

Permissions

Request permissions for this article.

Acknowledgments

We thank all of our co-workers and collaborators including T. Enami, R. Shibukawa, M. Funayama, M. Kawada, K. Goto, H. Houlden, E. Preza, and C. Okada for their technical support. We acknowledge P. Karagiannis for critical reading of the paper and N. Endo and R. Taniguchi for their administrative support. Funding: This work was funded in part by a grant from the iPS Cell Research Fund (S.Y.); the Program for Intractable Diseases Research utilizing Disease-specific iPS cells from the Japan Agency for Medical Research and Development (AMED) (H. Inoue); the Research Center Network for Realization of Regenerative Medicine from AMED (A.H., S.Y., and H. Inoue); Research Project for Practical Applications of Regenerative Medicine from AMED (A.O., T.E., and H. Inoue); Parkinson’s UK Senior Fellowship (F-0902) (T. Kunath); grant-in-aid for scientific research from the Japan Society for the Promotion of Science (15H04270; H. Ito, 15H05581; A.H.); and the Daiichi Sankyo Foundation of Life Science (H. Inoue). Author contributions: H. Inoue conceived the project; K.I. and H. Inoue designed the experiments; K.I., K.T., T.Y., T. Kondo, and S. Kitaoka performed cell culture, molecular experiments, and compound screen; A.W. performed single-cell analysis; K.I. and A. Tanaka performed animal experiments; S. Kaneko, T.A., and H. Ito performed human-sample analysis; N.O., M.H., and H.A. performed resequencing; K.I., A.W., T.Y., D.W., and H. Inoue analyzed the data; K.W., A.H., A.O., T. Kunath., S.W., T.E., T.F., H.N., K.H., H. Ichijo, J-.P.J., and S. Kaneko contributed reagents, materials, and analysis tools; Y.I., M.M., H.T., A. Tamaoka, H.F., K.M., K.O., and R.K. recruited patients; D.W., R.T., and S.Y. provided critical reading and scientific discussions; K.I. and H. Inoue wrote the paper. Competing interests: S.Y. is an unpaid scientific advisor to iPS Academia Japan. Kyoto University has filed patents related to this manuscript: PCT application PCT/JP2014/058142, entitled “Pluripotent stem cells for neuronal differentiation” with K.I. and H. Inoue as coinventors; and PCT application PCT/JP2016/050883, entitled “Agent for preventing and/or treating Amyotrophic lateral sclerosis” with K.I. and H. Inoue as coinventors. All other authors declare that they have no competing interests.

Authors

Affiliations

Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Department of Clinical Neuroscience, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8503, Japan.
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Kayoko Tsukita
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Knut Woltjen
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Hakubi Center for Advanced Research, Kyoto University, Kyoto 606-8501, Japan.
Takuya Yamamoto
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8507, Japan.
Akitsu Hotta
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8507, Japan.
Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Saitama 332-0012, Japan.
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Shiho Kitaoka
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Akito Tanaka
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Dai Watanabe
Department of Biological Sciences, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
Division of Neurology, Department of Internal Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
Hiroshi Takuma
Department of Neurology, Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan.
Akira Tamaoka
Department of Neurology, Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan.
MRC Centre for Regenerative Medicine, School of Biological Sciences, University of Edinburgh, 5 Little France Drive, Edinburgh EH16 4UU, UK.
Selina Wray
Department of Molecular Neuroscience, University College London Institute of Neurology, Queen Square, London WC1N 3BG, UK.
Department of Neurology, Kochi Medical School, Kochi University, Kochi 783-8505, Japan.
Takumi Era
Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto 860-0811, Japan.
Kouki Makioka
Department of Neurology, Gunma University Graduate School of Medicine, Maebashi 371-8511, Japan.
Koichi Okamoto
Geriatrics Research Institute and Hospital, Maebashi 371-0847, Japan.
Takao Fujisawa
Cell Signaling, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
Department of Medical Sciences, University of Miyazaki, Miyazaki 889-1601, Japan.
Cell Signaling, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
Hidenori Ichijo
Cell Signaling, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
Jean-Pierre Julien
Department of Psychiatry and Neurosciences, Research Centre of Mental Health Institute of Quebec (IUSMQ), Laval University, Québec, Canada.
Nanako Obata
Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo 156-8506, Japan.
Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo 156-8506, Japan.
Haruhiko Akiyama
Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo 156-8506, Japan.
Satoshi Kaneko
Department of Neurology, Kansai Medical University, Hirakata 573-1191, Japan.
Takashi Ayaki
Department of Neurology, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Department of Neurology, Wakayama Medical University, Kimiidera, Wakayama 641-8509, Japan.
Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8507, Japan.
Department of Neurology, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Shinya Yamanaka
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158, USA.
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.

Notes

*
Corresponding author. Email: [email protected]

Metrics & Citations

Metrics

Article Usage
Altmetrics

Citations

Export citation

Select the format you want to export the citation of this publication.

Cited by

  1. Stem cells in regenerative processes: Induced pluripotent stem cells, Regenerative Nephrology, (145-159), (2022).https://doi.org/10.1016/B978-0-12-823318-4.00030-5
    Crossref
  2. Traditional Chinese medicine compounds regulate autophagy for treating neurodegenerative disease: A mechanism review, Biomedicine & Pharmacotherapy, 133, (110968), (2021).https://doi.org/10.1016/j.biopha.2020.110968
    Crossref
  3. Drug Discovery in Induced Pluripotent Stem Cell Models, Reference Module in Biomedical Sciences, (2021).https://doi.org/10.1016/B978-0-12-820472-6.00049-9
    Crossref
  4. Towards Advanced iPSC-based Drug Development for Neurodegenerative Disease, Trends in Molecular Medicine, 27, 3, (263-279), (2021).https://doi.org/10.1016/j.molmed.2020.09.013
    Crossref
  5. Phenotyping Neurodegeneration in Human iPSCs, Annual Review of Biomedical Data Science, 4, 1, (83-100), (2021).https://doi.org/10.1146/annurev-biodatasci-092820-025214
    Crossref
  6. Small-Molecule Kinase Inhibitors for the Treatment of Nononcologic Diseases, Journal of Medicinal Chemistry, 64, 3, (1283-1345), (2021).https://doi.org/10.1021/acs.jmedchem.0c01511
    Crossref
  7. Induced pluripotent stem cells as models for Amyotrophic Lateral Sclerosis, iPSCs for Modeling Central Nervous System Disorders, (83-104), (2021).https://doi.org/10.1016/B978-0-323-85764-2.00001-6
    Crossref
  8. Electrophysiological Phenotype Characterization of Human iPSC‐Derived Neuronal Cell Lines by Means of High‐Density Microelectrode Arrays, Advanced Biology, 5, 3, (2000223), (2021).https://doi.org/10.1002/adbi.202000223
    Crossref
  9. Losing the Beat: Contribution of Purkinje Cell Firing Dysfunction to Disease, and Its Reversal, Neuroscience, 462, (247-261), (2021).https://doi.org/10.1016/j.neuroscience.2020.06.008
    Crossref
  10. Generation of Motor Neurons from Human ESCs/iPSCs Using Sendai Virus Vectors, Neural Reprogramming, (127-132), (2021).https://doi.org/10.1007/978-1-0716-1601-7_9
    Crossref
  11. See more
Loading...

View Options

Check Access

Log in to view the full text

AAAS ID LOGIN

AAAS login provides access to Science for AAAS Members, and access to other journals in the Science family to users who have purchased individual subscriptions.

Log in via OpenAthens.
Log in via Shibboleth.

More options

Register for free to read this article

As a service to the community, this article is available for free. Login or register for free to read this article.

View options

PDF format

Download this article as a PDF file

Download PDF

Full Text

FULL TEXT

Media

Figures

Multimedia

Tables

Share

Share

Share article link

Share on social media