1. Aberrant axon branching via Fos-B dysregulation in FUS-ALS motor neurons
- Author
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Fumiyoshi Fujishima, Jiro Kawada, Mitsuru Ishikawa, Hiroaki Mitsuhashi, Hideyuki Okano, Keiko Nakayama, Kensuke Ikeda, Mayumi Nishida, Shio Mitsuzawa, Rumiko Izumi, Matsuyuki Shirota, Ryo Funayama, Tomomi Shijo, Tetsuya Akiyama, Yasuo Kitajima, Shohei Kaneda, Masahiro Nogami, Hiroya Ono, Shion Osana, Yohei Okada, Naoki Suzuki, Takefumi Sone, Takayuki Kamei, Teruo Fujii, Satoru Morimoto, Ayumi Nishiyama, Masashi Aoki, Hitoshi Warita, Tadashi Nakagawa, Koki Fujimori, and Yoshiho Ikeuchi
- Subjects
Human-induced pluripotent stem cell (hiPSC)-derived motor neuron ,0301 basic medicine ,Research paper ,Nerve organoid ,Neurogenesis ,Induced Pluripotent Stem Cells ,Mutant ,Regulator ,Fos-B ,Axon branching ,Biology ,medicine.disease_cause ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,03 medical and health sciences ,0302 clinical medicine ,Fused in sarcoma (FUS) ,medicine ,Animals ,Humans ,Amyotrophic lateral sclerosis (ALS) ,RNA, Small Interfering ,Axon ,Amyotrophic lateral sclerosis ,Induced pluripotent stem cell ,Gene Editing ,Motor Neurons ,Messenger RNA ,Mutation ,Amyotrophic Lateral Sclerosis ,Lentivirus ,High-Throughput Nucleotide Sequencing ,Cell Differentiation ,General Medicine ,medicine.disease ,Phenotype ,Axons ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,RNA-Binding Protein FUS ,Proto-Oncogene Proteins c-fos - 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.
- Published
- 2019