title: ALS mutant FUS proteins are recruited into stress granules in induced pluripotent stem cell-derived motoneurons.
abstract: Patient-derived induced pluripotent stem cells (iPSCs) provide an opportunity to study human diseases mainly in those cases for which no suitable model systems are available. Here, we have taken advantage of in vitro iPSCs derived from patients affected by amyotrophic lateral sclerosis (ALS) and carrying mutations in the RNA-binding protein FUS to study the cellular behavior of the mutant proteins in the appropriate genetic background. Moreover, the ability to differentiate iPSCs into spinal cord neural cells provides an in vitro model mimicking the physiological conditions. iPSCs were derived from FUS(R514S) and FUS(R521C) patient fibroblasts, whereas in the case of the severe FUS(P525L) mutation, in which fibroblasts were not available, a heterozygous and a homozygous iPSC line were raised by TALEN-directed mutagenesis. We show that aberrant localization and recruitment of FUS into stress granules (SGs) is a prerogative of the FUS mutant proteins and occurs only upon induction of stress in both undifferentiated iPSCs and spinal cord neural cells. Moreover, we show that the incorporation into SGs is proportional to the amount of cytoplasmic FUS, strongly correlating with the cytoplasmic delocalization phenotype of the different mutants. Therefore, the available iPSCs represent a very powerful system for understanding the correlation between FUS mutations, the molecular mechanisms of SG formation and ALS ethiopathogenesis.
authors.affiliation: Center for Life Nano Science, Istituto Italiano di Tecnologia, Viale Regina Elena 291, Rome 00161, Italy Department of Biology and Biotechnology Charles Darwin, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy.
authors.forename: Jessica
authors.initials: J
authors.lastname: Lenzi
authors.affiliation: Center for Life Nano Science, Istituto Italiano di Tecnologia, Viale Regina Elena 291, Rome 00161, Italy Department of Biology and Biotechnology Charles Darwin, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: De Santis
authors.affiliation: Center for Life Nano Science, Istituto Italiano di Tecnologia, Viale Regina Elena 291, Rome 00161, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: de Turris
authors.affiliation: Department of Biology and Biotechnology Charles Darwin, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy.
authors.forename: Mariangela
authors.initials: M
authors.lastname: Morlando
authors.affiliation: Center for Life Nano Science, Istituto Italiano di Tecnologia, Viale Regina Elena 291, Rome 00161, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Laneve
authors.affiliation: Amyotrophic Lateral Sclerosis Center, Rita Levi Montalcini Department of Neuroscience, University of Turin, 10126 Turin, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Calvo
authors.affiliation: Dermatologia Chirurgica AOU Città della Salute e della Scienza, 10126 Turin, Italy.
authors.forename: Virginia
authors.initials: V
authors.lastname: Caliendo
authors.affiliation: Amyotrophic Lateral Sclerosis Center, Rita Levi Montalcini Department of Neuroscience, University of Turin, 10126 Turin, Italy.
authors.forename: Adriano
authors.initials: A
authors.lastname: Chiò
authors.affiliation: Department of Biology and Biotechnology Charles Darwin, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy alessandro.rosa@uniroma1.it irene.bozzoni@uniroma1.it.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Rosa
authors.affiliation: Center for Life Nano Science, Istituto Italiano di Tecnologia, Viale Regina Elena 291, Rome 00161, Italy Department of Biology and Biotechnology Charles Darwin, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy Institute Pasteur Fondazione Cenci-Bolognetti, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy alessandro.rosa@uniroma1.it irene.bozzoni@uniroma1.it.
authors.forename: Irene
authors.initials: I
authors.lastname: Bozzoni
mesh_terms: D021581:Active Transport, Cell Nucleus; D019943:Amino Acid Substitution; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D002454:Cell Differentiation; D002460:Cell Line; D003594:Cytoplasmic Granules / Q000378:metabolism / Q000473:pathology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000473:pathology; D008959:Models, Neurological; D009046:Motor Neurons / Q000378:metabolism* / Q000473:pathology*; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism*; D034702:RNA-Binding Protein FUS / Q000235:genetics / Q000378:metabolism*; D013116:Spinal Cord / Q000378:metabolism / Q000473:pathology; D013312:Stress, Physiological
chemical_list: C000599550:FUS protein, human; D050505:Mutant Proteins; D034702:RNA-Binding Protein FUS
keywords: ALS; FUS; TALE nucleases; iPSCs