title: Directly converted patient-specific induced neurons mirror the neuropathology of FUS with disrupted nuclear localization in amyotrophic lateral sclerosis.
abstract: BACKGROUND
  Mutations in the fused in sarcoma (FUS) gene have been linked to amyotrophic lateral sclerosis (ALS). ALS patients with FUS mutations exhibit neuronal cytoplasmic mislocalization of the mutant FUS protein. ALS patients' fibroblasts or induced pluripotent stem cell (iPSC)-derived neurons have been developed as models for understanding ALS-associated FUS (ALS-FUS) pathology; however, pathological neuronal signatures are not sufficiently present in the fibroblasts of patients, whereas the generation of iPSC-derived neurons from ALS patients requires relatively intricate procedures.
  
  
  RESULTS
  Here, we report the generation of disease-specific induced neurons (iNeurons) from the fibroblasts of patients who carry three different FUS mutations that were recently identified by direct sequencing and multi-gene panel analysis. The mutations are located at the C-terminal nuclear localization signal (NLS) region of the protein (p.G504Wfs*12, p.R495*, p.Q519E): two de novo mutations in sporadic ALS and one in familial ALS case. Aberrant cytoplasmic mislocalization with nuclear clearance was detected in all patient-derived iNeurons, and oxidative stress further induced the accumulation of cytoplasmic FUS in cytoplasmic granules, thereby recapitulating neuronal pathological features identified in mutant FUS (p.G504Wfs*12)-autopsied ALS patient. Importantly, such FUS pathological hallmarks of the patient with the p.Q519E mutation were only detected in patient-derived iNeurons, which contrasts to predominant FUS (p.Q519E) in the nucleus of both the transfected cells and patient-derived fibroblasts.
  
  
  CONCLUSIONS
  Thus, iNeurons may provide a more reliable model for investigating FUS mutations with disrupted NLS for understanding FUS-associated proteinopathies in ALS.
authors.affiliation: Department of Translational Medicine, Graduate School of Biomedical Science and Engineering, Hanyang University, Seoul, 133-792, Republic of Korea. lims7@hanyang.ac.kr.|Cell Therapy Center, Hanyang University Hospital, Seoul, 133-792, Republic of Korea. lims7@hanyang.ac.kr.
authors.forename: Su Min
authors.initials: SM
authors.lastname: Lim
authors.affiliation: Department of Neurology, Sheikh Khalifa Specialty Hospital, Ras Al Khaimah, United Arab Emirates. bbnorydan@gmail.com.
authors.forename: Won Jun
authors.initials: WJ
authors.lastname: Choi
authors.affiliation: Cell Therapy Center, Hanyang University Hospital, Seoul, 133-792, Republic of Korea. kiwook.oh@gmail.com.|Department of Neurology, College of Medicine, Hanyang University, Seoul, 133-792, Republic of Korea. kiwook.oh@gmail.com.
authors.forename: Ki-Wook
authors.initials: KW
authors.lastname: Oh
authors.affiliation: Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. ycxue@sun5.ibp.ac.cn.
authors.forename: Yuanchao
authors.initials: Y
authors.lastname: Xue
authors.affiliation: Cell Therapy Center, Hanyang University Hospital, Seoul, 133-792, Republic of Korea. ddg2102@gmail.com.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Choi
authors.affiliation: Cell Therapy Center, Hanyang University Hospital, Seoul, 133-792, Republic of Korea. whitehalf@hanmail.net.
authors.forename: Sung Hoon
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Cell Therapy Center, Hanyang University Hospital, Seoul, 133-792, Republic of Korea. nmy92@snu.ac.kr.
authors.forename: Minyeop
authors.initials: M
authors.lastname: Nahm
authors.affiliation: Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, 135-710, Republic of Korea. juliana.ye.kim@gmail.com.
authors.forename: Young-Eun
authors.initials: YE
authors.lastname: Kim
authors.affiliation: Korean Bioinformation Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-806, Republic of Korea. mack97hyuk@gmail.com.|Department of Bioinformatics, University of Sciences and Technology, Daejeon, 305-806, Republic of Korea. mack97hyuk@gmail.com.
authors.forename: Jinhyuk
authors.initials: J
authors.lastname: Lee
authors.affiliation: Cell Therapy Center, Hanyang University Hospital, Seoul, 133-792, Republic of Korea. my12163@hanmail.net.
authors.forename: Min-Young
authors.initials: MY
authors.lastname: Noh
authors.affiliation: Department of Brain and Cognitive Sciences, College of Natural Sciences, Seoul National University, Seoul, 110-744, Republic of Korea. seunglee@snu.ac.kr.
authors.forename: Seungbok
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, Seoul, 133-792, Republic of Korea. hwangsj@hanyang.ac.kr.
authors.forename: Sejin
authors.initials: S
authors.lastname: Hwang
authors.affiliation: Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, 135-710, Republic of Korea. changski@skku.edu.
authors.forename: Chang-Seok
authors.initials: CS
authors.lastname: Ki
authors.affiliation: Department of Cellular Molecular Medicine, University of California, La Jolla, San Diego, CA, 92093, USA. xdfu@ucsd.edu.
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Fu
authors.affiliation: Department of Translational Medicine, Graduate School of Biomedical Science and Engineering, Hanyang University, Seoul, 133-792, Republic of Korea. kimsh1@hanyang.ac.kr.|Cell Therapy Center, Hanyang University Hospital, Seoul, 133-792, Republic of Korea. kimsh1@hanyang.ac.kr.|Department of Neurology, College of Medicine, Hanyang University, Seoul, 133-792, Republic of Korea. kimsh1@hanyang.ac.kr.
authors.forename: Seung Hyun
authors.initials: SH
authors.lastname: Kim
mesh_terms: D000328:Adult; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics* / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000378:metabolism / Q000473:pathology; D005260:Female; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D008297:Male; D009046:Motor Neurons / Q000378:metabolism*; D050505:Mutant Proteins / Q000235:genetics; D009154:Mutation / Q000235:genetics*; D000069338:Neuropathology / Q000379:methods; D034702:RNA-Binding Protein FUS / Q000235:genetics / Q000378:metabolism*
chemical_list: D050505:Mutant Proteins; D034702:RNA-Binding Protein FUS