FUS is lost from nuclei and gained in neurites of motor neurons in a human stem cell model of VCP-related ALS

Harley, Jasmine; Hagemann, Cathleen; Serio, Andrea; Patani, Rickie

doi:10.1093/brain/awaa339

Cited by 15 publications

(25 citation statements)

References 10 publications

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“…Notably, we have previously demonstrated reduced nuclear-to-cytoplasmic ratio of SFPQ, FUS and TDP-43 proteins in VCP mutant hiPSC-derived neural precursors and/or motor neurons. 5 , 15 , 18 , 28 , 29 To contextualize these previous protein mislocalization findings with our current study, we next sought to examine the mechanistic relationship between RBP function and aberrant accumulation of IRTs in the cytoplasm, focusing on the interaction between SFPQ protein and SFPQ IRTs. We first performed RNA immunoprecipitation to demonstrate that the SFPQ protein physically interacts with SFPQ IRT ( Fig.…”

Section: Resultsmentioning

confidence: 88%

Aberrant cytoplasmic intron retention is a blueprint for RNA binding protein mislocalization in VCP-related amyotrophic lateral sclerosis

Tyzack

Neeves

Crerar

et al. 2021

Brain

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We recently described aberrantly increased cytoplasmic SFPQ intron-retaining transcripts (IRTs) and concurrent SFPQ protein mislocalization as new hallmarks of amyotrophic lateral sclerosis (ALS). However the generalizability and potential roles of cytoplasmic IRTs in health and disease remain unclear. Here, using time-resolved deep-sequencing of nuclear and cytoplasmic fractions of hiPSCs undergoing motor neurogenesis, we reveal that ALS-causing VCP gene mutations lead to compartment-specific aberrant accumulation of IRTs. Specifically, we identify >100 IRTs with increased cytoplasmic abundance in ALS samples. Furthermore, these aberrant cytoplasmic IRTs possess sequence-specific attributes and differential predicted binding affinity to RNA binding proteins (RBPs). Remarkably, TDP-43, SFPQ and FUS—RBPs known for nuclear-to-cytoplasmic mislocalization in ALS—abundantly and specifically bind to this aberrant cytoplasmic pool of IRTs, as opposed to any individual IRT. Our data are therefore consistent with a novel role for cytoplasmic IRTs in regulating compartment-specific protein abundance. This study provides new molecular insight into potential pathomechanisms underlying ALS and highlights aberrant cytoplasmic IRTs as potential therapeutic targets.

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Section: Resultsmentioning

confidence: 88%

Aberrant cytoplasmic intron retention is a blueprint for RNA binding protein mislocalization in VCP-related amyotrophic lateral sclerosis

Tyzack

Neeves

Crerar

et al. 2021

Brain

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“…Indeed neurite degeneration has been shown to occur upon oxidative stress through the cytoplasmic sequestration of two proteins (PRMT1 and Nd1-L) in in vitro models of FUS mutant-related ALS 35 . Furthermore TDP-43 mislocalization and aggregation has also been demonstrated in dystrophic neurites 1,28 , while we recently reported an increase in wild-type FUS within neuronal processes in VCP-mutant motor neurons 30 . Finally, a regulatory role for FUS has also been shown in synaptic formation and function [35][36][37][38] and aberrant FUS activity in the axonal compartment has been evidenced in a FUS mutant ALS mouse model 39 .…”

Section: Alsmentioning

confidence: 70%

“…CNN-based classifiers face challenges with interpretability and are not suited to specifically address the subcellular localisation of RBPs as previously described with conventional methods based on image segmentation 16,30 . Nevertheless here we showed that the phenotypes identified in the DAPI or III-tubulin fluorescent β images are indeed contained in the outlines of the nuclei and in the edges of the neurites respectively.…”

Section: Alsmentioning

confidence: 99%

Image-based deep learning reveals the responses of human motor neurons to stress and ALS

Verzat

Harley

Patani

et al. 2021

Preprint

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Although morphological attributes of cells and their substructures are recognized readouts of physiological or pathophysiological states, these have been relatively understudied in amyotrophic lateral sclerosis (ALS) research. In this study we integrate multichannel fluorescence high-content microscopy data with deep-learning imaging methods to reveal - directly from unsegmented images - novel neurite-associated morphological perturbations associated with (ALS-causing) VCP-mutant human motor neurons (MNs). Surprisingly, we reveal that previously unrecognized disease-relevant information is withheld in broadly used and often considered generic biological markers of nuclei (DAPI) and neurons (III-tubulin). Additionally, we identify changes within the information content of ALS-related RNA binding protein (RBP) immunofluorescence imaging that is captured in VCP-mutant MN cultures. Furthermore, by analyzing MN cultures exposed to different extrinsic stressors, we show that heat stress recapitulates key aspects of ALS. Our study therefore reveals disease-relevant information contained in a range of both generic and more specific fluorescent markers, and establishes the use of image-based deep learning methods for rapid, automated and unbiased testing of biological hypotheses.

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“…Interestingly, VCP mutations were identified in fALS patients [235]. Recently, FUS was found mislocalized in the cytoplasm of human induced pluripotent stem cells (iPSCs) derived from VCPmutant motoneurons [236,237]. Indeed, FUS cytoplasmic mislocalization under a diffuse form could be generalized in different ALS models, suggesting a broader role of FUS in the pathology.…”

Section: Nucleocytoplasmic Localizationmentioning

confidence: 99%

Amyotrophic Lateral Sclerosis Genes in Drosophila melanogaster

Layalle

They

Ourghani

et al. 2021

IJMS

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Amyotrophic lateral sclerosis (ALS) is a devastating adult-onset neurodegenerative disease characterized by the progressive degeneration of upper and lower motoneurons. Most ALS cases are sporadic but approximately 10% of ALS cases are due to inherited mutations in identified genes. ALS-causing mutations were identified in over 30 genes with superoxide dismutase-1 (SOD1), chromosome 9 open reading frame 72 (C9orf72), fused in sarcoma (FUS), and TAR DNA-binding protein (TARDBP, encoding TDP-43) being the most frequent. In the last few decades, Drosophila melanogaster emerged as a versatile model for studying neurodegenerative diseases, including ALS. In this review, we describe the different Drosophila ALS models that have been successfully used to decipher the cellular and molecular pathways associated with SOD1, C9orf72, FUS, and TDP-43. The study of the known fruit fly orthologs of these ALS-related genes yielded significant insights into cellular mechanisms and physiological functions. Moreover, genetic screening in tissue-specific gain-of-function mutants that mimic ALS-associated phenotypes identified disease-modifying genes. Here, we propose a comprehensive review on the Drosophila research focused on four ALS-linked genes that has revealed novel pathogenic mechanisms and identified potential therapeutic targets for future therapy.

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FUS is lost from nuclei and gained in neurites of motor neurons in a human stem cell model of VCP-related ALS

Cited by 15 publications

References 10 publications

Aberrant cytoplasmic intron retention is a blueprint for RNA binding protein mislocalization in VCP-related amyotrophic lateral sclerosis

Aberrant cytoplasmic intron retention is a blueprint for RNA binding protein mislocalization in VCP-related amyotrophic lateral sclerosis

Image-based deep learning reveals the responses of human motor neurons to stress and ALS

Amyotrophic Lateral Sclerosis Genes in Drosophila melanogaster

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