Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons

Tetsuya Akiyama, Naoki Suzuki, Mitsuru Ishikawa, Koki Fujimori, Takefumi Sone, Jiro Kawada, Ryo Funayama, Fumiyoshi Fujishima, Shio Mitsuzawa, Kensuke Ikeda, Hiroya Ono, Tomomi Shijo, Shion Osana, Matsuyuki Shirota, Tadashi Nakagawa, Yasuo Kitajima, Ayumi Nishiyama, Rumiko Izumi, Satoru Morimoto, Yohei OkadaTakayuki Kamei, Mayumi Nishida, Masahiro Nogami, Shohei Kaneda, Yoshiho Ikeuchi, Hiroaki Mitsuhashi, Keiko Nakayama, Teruo Fujii, Hitoshi Warita, Hideyuki Okano, Masashi Aoki

Research output: Contribution to journalArticle

Abstract

Background: The characteristic structure of motor neurons (MNs), particularly of the long axons, becomes damaged in the early stages of amyotrophic lateral sclerosis (ALS). However, the molecular pathophysiology of axonal degeneration remains to be fully elucidated. Method: Two sets of isogenic human-induced pluripotent stem cell (hiPSCs)-derived MNs possessing the single amino acid difference (p.H517D) in the fused in sarcoma (FUS) were constructed. By combining MN reporter lentivirus, MN specific phenotype was analyzed. Moreover, RNA profiling of isolated axons were conducted by applying the microfluidic devices that enable axon bundles to be produced for omics analysis. The relationship between the target gene, which was identified as a pathological candidate in ALS with RNA-sequencing, and the MN phenotype was confirmed by intervention with si-RNA or overexpression to hiPSCs-derived MNs and even in vivo. The commonality was further confirmed with other ALS-causative mutant hiPSCs-derived MNs and human pathology. Findings: We identified aberrant increasing of axon branchings in FUS-mutant hiPSCs-derived MN axons compared with isogenic controls as a novel phenotype. We identified increased level of Fos-B mRNA, the binding target of FUS, in FUS-mutant MNs. While Fos-B reduction using si-RNA or an inhibitor ameliorated the observed aberrant axon branching, Fos-B overexpression resulted in aberrant axon branching even in vivo. The commonality of those phenotypes was further confirmed with other ALS causative mutation than FUS. Interpretation: Analyzing the axonal fraction of hiPSC-derived MNs using microfluidic devices revealed that Fos-B is a key regulator of FUS-mutant axon branching. Fund: Japan Agency for Medical Research and development; Japanese Ministry of Education, Culture, Sports, Science and Technology Clinical Research, Innovation and Education Center, Tohoku University Hospital; Japan Intractable Diseases (Nanbyo) Research Foundation; the Kanae Foundation for the Promotion of Medical Science; and “Inochi-no-Iro” ALS research grant.

Original languageEnglish
Pages (from-to)362-378
Number of pages17
JournalEBioMedicine
Volume45
DOIs
Publication statusPublished - 2019 Jul 1

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Amyotrophic Lateral Sclerosis
Motor Neurons
Sarcoma
Neurons
Axons
Lab-On-A-Chip Devices
RNA
Phenotype
Induced Pluripotent Stem Cells
Research
Microfluidics
Japan
Education
RNA Sequence Analysis
Lentivirus
Organized Financing
Pathology
Sports
Stem cells
Biomedical Research

Keywords

  • Amyotrophic lateral sclerosis (ALS)
  • Axon branching
  • Fos-B
  • Fused in sarcoma (FUS)
  • Human-induced pluripotent stem cell (hiPSC)-derived motor neuron
  • Nerve organoid

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Akiyama, T., Suzuki, N., Ishikawa, M., Fujimori, K., Sone, T., Kawada, J., ... Aoki, M. (2019). Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons. EBioMedicine, 45, 362-378. https://doi.org/10.1016/j.ebiom.2019.06.013

Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons. / Akiyama, Tetsuya; Suzuki, Naoki; Ishikawa, Mitsuru; Fujimori, Koki; Sone, Takefumi; Kawada, Jiro; Funayama, Ryo; Fujishima, Fumiyoshi; Mitsuzawa, Shio; Ikeda, Kensuke; Ono, Hiroya; Shijo, Tomomi; Osana, Shion; Shirota, Matsuyuki; Nakagawa, Tadashi; Kitajima, Yasuo; Nishiyama, Ayumi; Izumi, Rumiko; Morimoto, Satoru; Okada, Yohei; Kamei, Takayuki; Nishida, Mayumi; Nogami, Masahiro; Kaneda, Shohei; Ikeuchi, Yoshiho; Mitsuhashi, Hiroaki; Nakayama, Keiko; Fujii, Teruo; Warita, Hitoshi; Okano, Hideyuki; Aoki, Masashi.

In: EBioMedicine, Vol. 45, 01.07.2019, p. 362-378.

Research output: Contribution to journalArticle

Akiyama, T, Suzuki, N, Ishikawa, M, Fujimori, K, Sone, T, Kawada, J, Funayama, R, Fujishima, F, Mitsuzawa, S, Ikeda, K, Ono, H, Shijo, T, Osana, S, Shirota, M, Nakagawa, T, Kitajima, Y, Nishiyama, A, Izumi, R, Morimoto, S, Okada, Y, Kamei, T, Nishida, M, Nogami, M, Kaneda, S, Ikeuchi, Y, Mitsuhashi, H, Nakayama, K, Fujii, T, Warita, H, Okano, H & Aoki, M 2019, 'Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons', EBioMedicine, vol. 45, pp. 362-378. https://doi.org/10.1016/j.ebiom.2019.06.013
Akiyama T, Suzuki N, Ishikawa M, Fujimori K, Sone T, Kawada J et al. Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons. EBioMedicine. 2019 Jul 1;45:362-378. https://doi.org/10.1016/j.ebiom.2019.06.013
Akiyama, Tetsuya ; Suzuki, Naoki ; Ishikawa, Mitsuru ; Fujimori, Koki ; Sone, Takefumi ; Kawada, Jiro ; Funayama, Ryo ; Fujishima, Fumiyoshi ; Mitsuzawa, Shio ; Ikeda, Kensuke ; Ono, Hiroya ; Shijo, Tomomi ; Osana, Shion ; Shirota, Matsuyuki ; Nakagawa, Tadashi ; Kitajima, Yasuo ; Nishiyama, Ayumi ; Izumi, Rumiko ; Morimoto, Satoru ; Okada, Yohei ; Kamei, Takayuki ; Nishida, Mayumi ; Nogami, Masahiro ; Kaneda, Shohei ; Ikeuchi, Yoshiho ; Mitsuhashi, Hiroaki ; Nakayama, Keiko ; Fujii, Teruo ; Warita, Hitoshi ; Okano, Hideyuki ; Aoki, Masashi. / Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons. In: EBioMedicine. 2019 ; Vol. 45. pp. 362-378.
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T1 - Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons

AU - Akiyama, Tetsuya

AU - Suzuki, Naoki

AU - Ishikawa, Mitsuru

AU - Fujimori, Koki

AU - Sone, Takefumi

AU - Kawada, Jiro

AU - Funayama, Ryo

AU - Fujishima, Fumiyoshi

AU - Mitsuzawa, Shio

AU - Ikeda, Kensuke

AU - Ono, Hiroya

AU - Shijo, Tomomi

AU - Osana, Shion

AU - Shirota, Matsuyuki

AU - Nakagawa, Tadashi

AU - Kitajima, Yasuo

AU - Nishiyama, Ayumi

AU - Izumi, Rumiko

AU - Morimoto, Satoru

AU - Okada, Yohei

AU - Kamei, Takayuki

AU - Nishida, Mayumi

AU - Nogami, Masahiro

AU - Kaneda, Shohei

AU - Ikeuchi, Yoshiho

AU - Mitsuhashi, Hiroaki

AU - Nakayama, Keiko

AU - Fujii, Teruo

AU - Warita, Hitoshi

AU - Okano, Hideyuki

AU - Aoki, Masashi

PY - 2019/7/1

Y1 - 2019/7/1

N2 - Background: The characteristic structure of motor neurons (MNs), particularly of the long axons, becomes damaged in the early stages of amyotrophic lateral sclerosis (ALS). However, the molecular pathophysiology of axonal degeneration remains to be fully elucidated. Method: Two sets of isogenic human-induced pluripotent stem cell (hiPSCs)-derived MNs possessing the single amino acid difference (p.H517D) in the fused in sarcoma (FUS) were constructed. By combining MN reporter lentivirus, MN specific phenotype was analyzed. Moreover, RNA profiling of isolated axons were conducted by applying the microfluidic devices that enable axon bundles to be produced for omics analysis. The relationship between the target gene, which was identified as a pathological candidate in ALS with RNA-sequencing, and the MN phenotype was confirmed by intervention with si-RNA or overexpression to hiPSCs-derived MNs and even in vivo. The commonality was further confirmed with other ALS-causative mutant hiPSCs-derived MNs and human pathology. Findings: We identified aberrant increasing of axon branchings in FUS-mutant hiPSCs-derived MN axons compared with isogenic controls as a novel phenotype. We identified increased level of Fos-B mRNA, the binding target of FUS, in FUS-mutant MNs. While Fos-B reduction using si-RNA or an inhibitor ameliorated the observed aberrant axon branching, Fos-B overexpression resulted in aberrant axon branching even in vivo. The commonality of those phenotypes was further confirmed with other ALS causative mutation than FUS. Interpretation: Analyzing the axonal fraction of hiPSC-derived MNs using microfluidic devices revealed that Fos-B is a key regulator of FUS-mutant axon branching. Fund: Japan Agency for Medical Research and development; Japanese Ministry of Education, Culture, Sports, Science and Technology Clinical Research, Innovation and Education Center, Tohoku University Hospital; Japan Intractable Diseases (Nanbyo) Research Foundation; the Kanae Foundation for the Promotion of Medical Science; and “Inochi-no-Iro” ALS research grant.

AB - Background: The characteristic structure of motor neurons (MNs), particularly of the long axons, becomes damaged in the early stages of amyotrophic lateral sclerosis (ALS). However, the molecular pathophysiology of axonal degeneration remains to be fully elucidated. Method: Two sets of isogenic human-induced pluripotent stem cell (hiPSCs)-derived MNs possessing the single amino acid difference (p.H517D) in the fused in sarcoma (FUS) were constructed. By combining MN reporter lentivirus, MN specific phenotype was analyzed. Moreover, RNA profiling of isolated axons were conducted by applying the microfluidic devices that enable axon bundles to be produced for omics analysis. The relationship between the target gene, which was identified as a pathological candidate in ALS with RNA-sequencing, and the MN phenotype was confirmed by intervention with si-RNA or overexpression to hiPSCs-derived MNs and even in vivo. The commonality was further confirmed with other ALS-causative mutant hiPSCs-derived MNs and human pathology. Findings: We identified aberrant increasing of axon branchings in FUS-mutant hiPSCs-derived MN axons compared with isogenic controls as a novel phenotype. We identified increased level of Fos-B mRNA, the binding target of FUS, in FUS-mutant MNs. While Fos-B reduction using si-RNA or an inhibitor ameliorated the observed aberrant axon branching, Fos-B overexpression resulted in aberrant axon branching even in vivo. The commonality of those phenotypes was further confirmed with other ALS causative mutation than FUS. Interpretation: Analyzing the axonal fraction of hiPSC-derived MNs using microfluidic devices revealed that Fos-B is a key regulator of FUS-mutant axon branching. Fund: Japan Agency for Medical Research and development; Japanese Ministry of Education, Culture, Sports, Science and Technology Clinical Research, Innovation and Education Center, Tohoku University Hospital; Japan Intractable Diseases (Nanbyo) Research Foundation; the Kanae Foundation for the Promotion of Medical Science; and “Inochi-no-Iro” ALS research grant.

KW - Amyotrophic lateral sclerosis (ALS)

KW - Axon branching

KW - Fos-B

KW - Fused in sarcoma (FUS)

KW - Human-induced pluripotent stem cell (hiPSC)-derived motor neuron

KW - Nerve organoid

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