@article{288bf382bbf940149de9bb8fe5f9bed9,
title = "Reduced PHOX2B stability causes axonal growth impairment in motor neurons with TARDBP mutations",
abstract = "Amyotrophic lateral sclerosis (ALS) is an adult-onset incurable motor neuron (MN) disease. The reasons for selective MN vulnerability in ALS are unknown. Axonal pathology is among the earliest signs of ALS. We searched for novel modulatory genes in human MN axon shortening affected by TARDBP mutations. In transcriptome analysis of RNA present in the axon compartment of human-derived induced pluripotent stem cell (iPSC)-derived MNs, PHOX2B (paired-like homeobox protein 2B) showed lower expression in TARDBP mutant axons, which was consistent with axon qPCR and in situ hybridization. PHOX2B mRNA stability was reduced in TARDBP mutant MNs. Furthermore, PHOX2B knockdown reduced neurite length in human MNs. Finally, phox2b knockdown in zebrafish induced short spinal axons and impaired escape response. PHOX2B is known to be highly express in other types of neurons maintained after ALS progression. Collectively, TARDBP mutations induced loss of axonal resilience, which is an important ALS-related phenotype mediated by PHOX2B downregulation.",
keywords = "TAR-DNA binding protein (TARDBP), amyotrophic lateral sclerosis (ALS), human-induced pluripotent stem cell (hiPSC)-derived motor neurons, neurite length",
author = "Shio Mitsuzawa and Naoki Suzuki and Tetsuya Akiyama and Mitsuru Ishikawa and Takefumi Sone and Jiro Kawada and Ryo Funayama and Matsuyuki Shirota and Hiroaki Mitsuhashi and Satoru Morimoto and Kensuke Ikeda and Tomomi Shijo and Akiyuki Ohno and Naoko Nakamura and Hiroya Ono and Risako Ono and Shion Osana and Tadashi Nakagawa and Ayumi Nishiyama and Rumiko Izumi and Shohei Kaneda and Yoshiho Ikeuchi and Keiko Nakayama and Teruo Fujii and Hitoshi Warita and Hideyuki Okano and Masashi Aoki",
note = "Funding Information: The authors thank N. Sugeno, T. Hasegawa, Y. Takai, T. Misu, and M. Kato of the Department of Neurology, Tohoku University, for advice regarding this research; M. Suzuki, N. Shimakura, H. Shigihara, and A. Machii of the Department of Neurology, Tohoku University, for their technical assistance; K. Kuroda, M. Kikuchi, and M. Nakagawa of the Division of Cell Proliferation, Tohoku University, and A. Tabe of Subio for assistance with the RNA-seq procedure; S. Koyama, T. Kato, and Y. Suzuki for ongoing care of the G376D proband; H. Inoue, S. Yamanaka, and M. Nakagawa (Kyoto University) for donating hiPSC clones 409B2, 201B7, and CiRA00026; S. Nakamura and F. Ozawa for cells shipping and maintenance in Department of Physiology, Keio University; T. Niihori and Y. Aoki for genetic analysis; T. Kamei, T. Yamashita, and T. Matsuo of Takeda Pharmaceutical Company for their gift of TARDBP mutant isogenic iPSCs; A. Miyawaki and H. Miyoshi for donating lentivirus vectors; and University of California, San Diego ([UCSD], San Diego, USA) for permission to use their RFP; H. Momma for statistical advice; Y. Akiyama for the illustrations. We also thank the Biomedical Research Unit of Tohoku University Hospital for providing the locations to maintain hiPSCs. Zebrafish line Tg [hb9:Venus] and wild-type strain RIKEN WT were obtained from National BioResource Project, Zebrafish Core Institution, Japan. Sanger sequencing of zebrafish phox2b DNA constructs was performed by the Support Center for Medical Research and Education, Tokai University. This work was supported by JSPS KAKENHI grant No. JP20K16593. The authors would also like to thank Enago (www.enago.jp) for the English language review. Funding Information: The authors thank N. Sugeno, T. Hasegawa, Y. Takai, T. Misu, and M. Kato of the Department of Neurology, Tohoku University, for advice regarding this research; M. Suzuki, N. Shimakura, H. Shigihara, and A. Machii of the Department of Neurology, Tohoku University, for their technical assistance; K. Kuroda, M. Kikuchi, and M. Nakagawa of the Division of Cell Proliferation, Tohoku University, and A. Tabe of Subio for assistance with the RNA-seq procedure; S. Koyama, T. Kato, and Y. Suzuki for ongoing care of the G376D proband; H. Inoue, S. Yamanaka, and M. Nakagawa (Kyoto University) for donating hiPSC clones 409B2, 201B7, and CiRA00026; S. Nakamura and F. Ozawa for cells shipping and maintenance in Department of Physiology, Keio University; T. Niihori and Y. Aoki for genetic analysis; T. Kamei, T. Yamashita, and T. Matsuo of Takeda Pharmaceutical Company for their gift of TARDBP mutant isogenic iPSCs; A. Miyawaki and H. Miyoshi for donating lentivirus vectors; and University of California, San Diego ([UCSD], San Diego, USA) for permission to use their RFP; H. Momma for statistical advice; Y. Akiyama for the illustrations. We also thank the Biomedical Research Unit of Tohoku University Hospital for providing the locations to maintain hiPSCs. Zebrafish line Tg [hb9:Venus] and wild-type strain RIKEN WT were obtained from National BioResource Project, Zebrafish Core Institution, Japan. Sanger sequencing of zebrafish phox2b DNA constructs was performed by the Support Center for Medical Research and Education, Tokai University. This work was supported by JSPS KAKENHI grant No. JP20K16593 . The authors would also like to thank Enago ( www.enago.jp ) for the English language review. Publisher Copyright: {\textcopyright} 2021 The Authors",
year = "2021",
month = jun,
day = "8",
doi = "10.1016/j.stemcr.2021.04.021",
language = "English",
volume = "16",
pages = "1527--1541",
journal = "Stem Cell Reports",
issn = "2213-6711",
publisher = "Cell Press",
number = "6",
}
