The fused in sarcoma protein forms cytoplasmic aggregates in motor neurons derived from integration-free induced pluripotent stem cells generated from a patient with familial amyotrophic lateral sclerosis carrying the FUS-P525L mutation

Liu, Xinxiu; Chen, Jiayu; Liu, WenChao; Li, Yong; Chen, Qi; Liu, Tao; Gao, Shaorong; Deng, Min

doi:10.1007/s10048-015-0448-y

Cited by 31 publications

(27 citation statements)

References 35 publications

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“…In young, 21 days differentiated MNs, FUS was mainly localized in nuclei, except for the most aggressive mutation in which FUS was already translocated into the cytoplasm. In mature 42 days differentiated MN from control cell lines, FUS was not only localized in the nuclei (Japtok et al, 2015; Lenzi et al, 2015; Liu et al, 2015) but was also detectable as small punctae along the neurites. This observation was very similar to the FUS localization along neurites and within the presynaptic compartment in hippocampal neurons (Schoen et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

“…Only a few studies using hiPS cell lines expressing different mFUS variants are published up to now showing a cytoplasmatic distribution of FUS in mFUS hiPSC derived spinal MN (Lenzi et al, 2015; Liu et al, 2015; Ichiyanagi et al, 2016; Naujock et al, 2016) and an enhanced recruitment of FUS into SGs after the induction of cellular stress. Similarly, in hiPSC-derived cortical neurons expressing different mFUS variants (a mild mutation R521C and an aggressive mutation R495QfsX527), cytoplasmic FUS inclusions were spontaneously formed and were dependent on both, the severity of the mutation and neuronal age.…”

Section: Introductionmentioning

confidence: 99%

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FUS Mislocalization and Vulnerability to DNA Damage in ALS Patients Derived hiPSCs and Aging Motoneurons

Higelin

Demestre

Putz

et al. 2016

Front. Cell. Neurosci.

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Mutations within the FUS gene (Fused in Sarcoma) are known to cause Amyotrophic Lateral Sclerosis (ALS), a neurodegenerative disease affecting upper and lower motoneurons. The FUS gene codes for a multifunctional RNA/DNA-binding protein that is primarily localized in the nucleus and is involved in cellular processes such as splicing, translation, mRNA transport and DNA damage response. In this study, we analyzed pathophysiological alterations associated with ALS related FUS mutations (mFUS) in human induced pluripotent stem cells (hiPSCs) and hiPSC derived motoneurons. To that end, we compared cells carrying a mild or severe mFUS in physiological- and/or stress conditions as well as after induced DNA damage. Following hyperosmolar stress or irradiation, mFUS hiPS cells recruited significantly more cytoplasmatic FUS into stress granules accompanied by impaired DNA-damage repair. In motoneurons wild-type FUS was localized in the nucleus but also deposited as small punctae within neurites. In motoneurons expressing mFUS the protein was additionally detected in the cytoplasm and a significantly increased number of large, densely packed FUS positive stress granules were seen along neurites. The amount of FUS mislocalization correlated positively with both the onset of the human disease (the earlier the onset the higher the FUS mislocalization) and the maturation status of the motoneurons. Moreover, even in non-stressed post-mitotic mFUS motoneurons clear signs of DNA-damage could be detected. In summary, we found that the susceptibility to cell stress was higher in mFUS hiPSCs and hiPSC derived motoneurons than in controls and the degree of FUS mislocalization correlated well with the clinical severity of the underlying ALS related mFUS. The accumulation of DNA damage and the cellular response to DNA damage stressors was more pronounced in post-mitotic mFUS motoneurons than in dividing hiPSCs suggesting that mFUS motoneurons accumulate foci of DNA damage, which in turn might be directly linked to neurodegeneration.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

FUS Mislocalization and Vulnerability to DNA Damage in ALS Patients Derived hiPSCs and Aging Motoneurons

Higelin

Demestre

Putz

et al. 2016

Front. Cell. Neurosci.

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“…In 2008, the first ALS patient iPSC-derived motor neurons (MNs) were generated 4 . Since then, many ALS iPSC studies have been reported,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 and this technology is leading to new findings and therapeutic candidates for ALS.…”

Section: Introductionmentioning

confidence: 99%

Simple Derivation of Spinal Motor Neurons from ESCs/iPSCs Using Sendai Virus Vectors

Goto

Imamura

Komatsu

et al. 2017

Molecular Therapy - Methods & Clinical Development

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Amyotrophic lateral sclerosis (ALS) is a progressive and fatal degenerative disorder of motor neurons (MNs). Embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs) now help us to understand the pathomechanisms of ALS via disease modeling. Various methods to differentiate ESCs/iPSCs into MNs by the addition of signaling molecules have been reported. However, classical methods require multiple steps, and newer simple methods using the transduction of transcription factors run the risk of genomic integration of the vector genes. Heterogeneity of the expression levels of the transcription factors also remains an issue. Here we describe a novel approach for differentiating human and mouse ESCs/iPSCs into MNs using a single Sendai virus vector encoding three transcription factors, LIM/homeobox protein 3, neurogenin 2, and islet-1, which are integration free. This single-vector method, generating HB9-positive cells on day 2 from human iPSCs, increases the ratio of MNs to neurons compared to the use of three separate Sendai virus vectors. In addition, the MNs derived via this method from iPSCs of ALS patients and model mice display disease phenotypes. This simple approach significantly reduces the efforts required to generate MNs, and it provides a useful tool for disease modeling.

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“…In addition, the use of recently developed induced pluripotent stem cell (iPSC) technologies also enables understanding of the disease pathogenesis ( Mattis and Svendsen, 2011 , Okano and Yamanaka, 2014 ). Indeed, iPSCs have been generated from ALS patients with mutations in SOD1 ( Chestkov and Vasilieva, 2014 , Dimos et al., 2008 ), TDP-43 ( Bilican et al., 2012 , Egawa et al., 2012 ), C9ORF72 ( Almeida et al., 2013 , Sareen et al., 2013 ) and recent publications of FUS ( Lenzi et al., 2015 , Liu et al., 2015 , Di Salvio et al., 2015 ) suggest a useful tool for pursuing the cellular pathogenesis and mechanism underlying FALS.…”

Section: Introductionmentioning

confidence: 99%

Establishment of In Vitro FUS-Associated Familial Amyotrophic Lateral Sclerosis Model Using Human Induced Pluripotent Stem Cells

et al. 2016

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SummaryAmyotrophic lateral sclerosis (ALS) is a late-onset motor neuron disorder. Although its neuropathology is well understood, the cellular and molecular mechanisms are yet to be elucidated due to limitations in the currently available human genetic data. In this study, we generated induced pluripotent stem cells (iPSC) from two familial ALS (FALS) patients with a missense mutation in the fused-in sarcoma (FUS) gene carrying the heterozygous FUS H517D mutation, and isogenic iPSCs with the homozygous FUS H517D mutation by genome editing technology. These cell-derived motor neurons mimicked several neurodegenerative phenotypes including mis-localization of FUS into cytosolic and stress granules under stress conditions, and cellular vulnerability. Moreover, exon array analysis using motor neuron precursor cells (MPCs) combined with CLIP-seq datasets revealed aberrant gene expression and/or splicing pattern in FALS MPCs. These results suggest that iPSC-derived motor neurons are a useful tool for analyzing the pathogenesis of human motor neuron disorders.

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The fused in sarcoma protein forms cytoplasmic aggregates in motor neurons derived from integration-free induced pluripotent stem cells generated from a patient with familial amyotrophic lateral sclerosis carrying the FUS-P525L mutation

Cited by 31 publications

References 35 publications

FUS Mislocalization and Vulnerability to DNA Damage in ALS Patients Derived hiPSCs and Aging Motoneurons

FUS Mislocalization and Vulnerability to DNA Damage in ALS Patients Derived hiPSCs and Aging Motoneurons

Simple Derivation of Spinal Motor Neurons from ESCs/iPSCs Using Sendai Virus Vectors

Establishment of In Vitro FUS-Associated Familial Amyotrophic Lateral Sclerosis Model Using Human Induced Pluripotent Stem Cells

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