Stepwise acquirement of hallmark neuropathology in FUS-ALS iPSC models depends on mutation type and neuronal aging

Japtok, Julia; Lojewski, Xenia; Naumann, Marcel; Klingenstein, Moritz; Reinhardt, Peter; Sterneckert, Jared; Putz, Stefan; Demestre, Maria; Boeckers, Tobias M.; Ludolph, Albert C.; Liebau, Stefan; Storch, Alexander; Hermann, Andreas

doi:10.1016/j.nbd.2015.07.017

Cited by 61 publications

(71 citation statements)

References 47 publications

(92 reference statements)

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“…FUS1 harbored a “mild” missense mutation (R521C) with a late onset (57 years old), family history of ALS (Japtok et al, 2015), and a disease duration of 7 months. Cell line FUS2 carried the “malign” novel 1 bp deletion c.1483delC leading to a frameshift and the translation of 33 “new” amino acids before the STOP codon (R495QfsX527; Belzil et al, 2012; Japtok et al, 2015; Lenzi et al, 2015). This mutation presented with a juvenile ALS onset (27 years old) and death 16 months after onset.…”

Section: Resultsmentioning

confidence: 99%

“…Moreover, we corrected the mutation of the FUS2 line by CRISPR technology ( Supplementary Figure S2 ) to test for mutation specific phenotypes (isogenic CNTL cell line R495QfsX527 c.1483insC ) ( Table 1 ). All established hiPS cell lines (Asp502Thrfs ∗ 27 is shown as an example) were checked for the expression of characteristic pluripotency markers including the nuclear factors OCT4, SOX2, NANOG and SSEA-4,TRA-1-60 and TRA-1-81 ( Supplementary Figures S1A,B or as previously published (Japtok et al, 2015; Naujock et al, 2016). To confirm the ability of iPSCs to differentiate into ectoderm, endoderm, and mesoderm, specific markers were analyzed by immunocytochemistry including tubulin-ß-III, actinin, and AFP ( Supplementary Figure S1C ).…”

Section: Resultsmentioning

confidence: 99%

“…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: 98%

“…We generated a novel iPS cell line from a juvenile ALS patient presenting with a frame shift FUS mutation in the RGG-rich region Asp502Thrfs ∗ 27 (Hübers et al, 2015) and compared it with our previously described cell lines R521C and R495QfsX527 (Japtok et al, 2015). In iPS cells under physiological in vitro conditions FUS was only mislocalized in the cell line Asp502Thrfs ∗ 27 but not in the other cell lines in which mFUS remained mainly nuclear.…”

Section: Discussionmentioning

confidence: 99%

“…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. Furthermore, the severity of the FUS mutation determined the cell vulnerability to oxidative stress (Japtok et al, 2015). In addition to the two previously described mutations, we generated and differentiated an additional hiPSC line from a juvenile ALS-patient with a severe de novo FUS mutation Asp502Thrfs ∗ 27 (Hübers et al, 2015).…”

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: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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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Studying Human Neurological Disorders Using Induced Pluripotent Stem Cells: From 2D Monolayer to 3D Organoid and Blood Brain Barrier Models

Logan

Arzua

Canfield

et al. 2019

Comprehensive Physiology

103

145

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Neurological disorders have emerged as a predominant healthcare concern in recent years due to their severe consequences on quality of life and prevalence throughout the world. Understanding the underlying mechanisms of these diseases and the interactions between different brain cell types is essential for the development of new therapeutics. Induced pluripotent stem cells (iPSCs) are invaluable tools for neurological disease modeling, as they have unlimited self-renewal and differentiation capacity. Mounting evidence shows: 1) various brain cells can be generated from iPSCs in 2-dimensional (2D) monolayer cultures; 2) further advances in 3D culture systems have led to the differentiation of iPSCs into organoids with multiple brain cell types and specific brain regions. These 3D organoids have gained widespread attention as in vitro tools to recapitulate complex features of the brain, and 3) Complex interactions between iPSC-derived brain cell types can recapitulate physiological and pathological conditions of blood-brain barrier (BBB). As iPSCs can be generated from diverse patient populations, researchers have effectively applied 2D, 3D and BBB models to recapitulate genetically complex neurological disorders and reveal novel insights into molecular and genetic mechanisms of neurological disorders. In this review, we describe recent progress in the generation of 2D, 3D and BBB models from iPSCs and further discuss their limitations, advantages, and future ventures. This review also covers the current status of applications of 2D, 3D and BBB models in drug screening, precision medicine, and modeling a wide range of neurological diseases (e.g., neurodegenerative diseases, neurodevelopmental disorders, brain injury, and neuropsychiatric disorders).

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Dysregulated axonal RNA translation in amyotrophic lateral sclerosis

Yasuda

Mili

2016

WIREs RNA

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Amyotrophic lateral sclerosis (ALS) is an adult‐onset motor neuron disease that has been associated with a diverse array of genetic changes. Prominent among these are mutations in RNA‐binding proteins (RBPs) or repeat expansions that give rise to toxic RNA species. RBPs are additionally central components of pathologic aggregates that constitute a disease hallmark, suggesting that dysregulation of RNA metabolism underlies disease progression. In the context of neuronal physiology, transport of RNAs and localized RNA translation in axons are fundamental to neuronal survival and function. Several lines of evidence suggest that axonal RNA translation is a central process perturbed by various pathogenic events associated with ALS. Dysregulated translation of specific RNA groups could underlie feedback effects that connect and reinforce disease manifestations. Among such candidates are RNAs encoding proteins involved in the regulation of microtubule dynamics. Further understanding of axonally dysregulated RNA targets and of the feedback mechanisms they induce could provide useful therapeutic insights. WIREs RNA 2016, 7:589–603. doi: 10.1002/wrna.1352For further resources related to this article, please visit the WIREs website.

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Stepwise acquirement of hallmark neuropathology in FUS-ALS iPSC models depends on mutation type and neuronal aging

Cited by 61 publications

References 47 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

Studying Human Neurological Disorders Using Induced Pluripotent Stem Cells: From 2D Monolayer to 3D Organoid and Blood Brain Barrier Models

Dysregulated axonal RNA translation in amyotrophic lateral sclerosis

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