FUS pathology in ALS is linked to alterations in multiple ALS-associated proteins and rescued by drugs stimulating autophagy

Marrone, Lara; Drexler, Hannes C.A.; Wang, Jie; Tripathi, Priyanka; Distler, Tania; Heisterkamp, Patrick; Anderson, Eric N.; Kour, Sukhleen; Moraiti, Anastasia; Maharana, Shovamayee; Bhatnagar, Rajat; Belgard, T. Grant; Tripathy, Vadreenath; Kalmbach, Norman; Hosseinzadeh, Zohreh; Crippa, V.; Abo-Rady, Masin; Wegner, Florian; Poletti, Angelo; Troost, Dirk; Aronica, Eleonora; Busskamp, Volker; Weis, Joachim; Pandey, Udai Bhan; Hyman, Anthony; Alberti, Simon; Goswami, Anand; Sterneckert, Jared

doi:10.1007/s00401-019-01998-x

Cited by 94 publications

(92 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One possibility is that different FG-Nups might stabilize the liquid state when combined together. Analogues for this hypothesis are ubiquitous in the literature, e.g., the inhibition of amyloid-β plaques or aggregates by α-synuclein (Bachhuber et al, 2015) or mixing of the intrinsically disordered protein FUS with EWS or TAF15 (Marrone et al, 2019). In the future, more complex device designs that have additional mixers for on-chip preparation of different FG-Nups may address this question.…”

Section: Resultsmentioning

confidence: 99%

The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes

Celetti

Paci

Caria

et al. 2019

Journal of Cell Biology

104

112

View full text Add to dashboard Cite

Nuclear pore complexes (NPCs) regulate all cargo traffic across the nuclear envelope. The transport conduit of NPCs is highly enriched in disordered phenylalanine/glycine-rich nucleoporins (FG-Nups), which form a permeability barrier of still elusive and highly debated molecular structure. Here we present a microfluidic device that triggered liquid-to-liquid phase separation of FG-Nups, which yielded droplets that showed typical properties of a liquid state. On the microfluidic chip, droplets were perfused with different transport-competent or -incompetent cargo complexes, and then the permeability barrier properties of the droplets were optically interrogated. We show that the liquid state mimics permeability barrier properties of the physiological nuclear transport pathway in intact NPCs in cells: that is, inert cargoes ranging from small proteins to large capsids were excluded from liquid FG-Nup droplets, but functional import complexes underwent facilitated import into droplets. Collectively, these data provide an experimental model of how NPCs can facilitate fast passage of cargoes across an order of magnitude in cargo size.

show abstract

Section: Resultsmentioning

confidence: 99%

The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes

Celetti

Paci

Caria

et al. 2019

Journal of Cell Biology

104

112

View full text Add to dashboard Cite

show abstract

“…Finally, our study provides an additional proof of concept that modulating autophagy may be of therapeutic interest in ALS/FTD. Indeed, previous studies have shown that stimulating autophagy is beneficial in SOD1, FUS, and TDP‐43 cell or animal models of ALS (Hetz et al , ; Crippa et al , ; Wang et al , ; Castillo et al , ; Barmada et al , ; Perera et al , ; Marrone et al , , ). In agreement with these works, we found that compounds known to induce autophagy, notably promethazine, are able to bypass the reduced expression of C9ORF72 and can promote the clearance of DPR proteins, thus reducing their toxicity.…”

Section: Discussionmentioning

confidence: 99%

“…Promethazine is a sedative and antihistamine drug with antipsychotic effects that belongs to a class of phenothiazine derivatives known to activate autophagy in neuronal cell cultures (Tsvetkov et al , ). Interestingly, these compounds ameliorate neuronal cell dysfunctions and prevent neuronal cell death in SOD1, TDP‐43, and FUS cellular and animal models of ALS (Barmada et al , ; Patten et al , ; Marrone et al , , ). Furthermore, pimozide, a phenothiazine derivative related to promethazine, displayed some promising effects in a short randomized clinical trial of sporadic ALS (Patten et al , ).…”

Section: Discussionmentioning

confidence: 99%

Reduced autophagy upon C9ORF72 loss synergizes with dipeptide repeat protein toxicity in G4C2 repeat expansion disorders

et al. 2020

View full text Add to dashboard Cite

Expansion of G4C2 repeats within the C9ORF72 gene is the most common cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Such repeats lead to decreased expression of the autophagy regulator C9ORF72 protein. Furthermore, sense and antisense repeats are translated into toxic dipeptide repeat (DPR) proteins. It is unclear how these repeats are translated, and in which way their translation and the reduced expression of C9ORF72 modulate repeat toxicity. Here, we found that sense and antisense repeats are translated upon initiation at canonical AUG or near‐cognate start codons, resulting in polyGA‐, polyPG‐, and to a lesser degree polyGR‐DPR proteins. However, accumulation of these proteins is prevented by autophagy. Importantly, reduced C9ORF72 levels lead to suboptimal autophagy, thereby impairing clearance of DPR proteins and causing their toxic accumulation, ultimately resulting in neuronal cell death. Of clinical importance, pharmacological compounds activating autophagy can prevent neuronal cell death caused by DPR proteins accumulation. These results suggest the existence of a double‐hit pathogenic mechanism in ALS/FTD, whereby reduced expression of C9ORF72 synergizes with DPR protein accumulation and toxicity.

show abstract

“…The inhibition of autophagy was 286 observed when over-expressing altFUS alone, absent when over-expressing FUS alone (wild-type 287 or ALS-associated R495x mutant) and reconstituted when co-expressing FUS and altFUS. This 288 demonstrates that the inhibition of autophagy, previously described in FUS-linked ALS 11,21 , has 289 been incorrectly associated to the FUS protein. AltFUS inhibits autophagy.…”

mentioning

confidence: 91%

“…The 53 protein is involved in RNA processing, DNA repair and cellular proliferation, although its functions 54 are not precisely elucidated 9 . Most of the mutations associated with neurodegenerative diseases 55 alter FUS NLS [11][12][13] . More recently, mutations in FUS 3'UTR were described in ALS patients and 56 linked to an increased level of FUS mRNA and protein [14][15][16] .…”

mentioning

confidence: 99%

FUSgene is dual-coding with both proteins united in FUS-mediated toxicity

Brunet

Jacques

Nassari

et al. 2019

Preprint

View full text Add to dashboard Cite

MAIN 44Amyotrophic Lateral Sclerosis (ALS) is a fatal motor neuron degenerative disease 1-3 characterized 45 by a progressive loss of motor neurons in the brain, brainstem and spinal cord 1 . Most cases are 46 sporadic (sALS), although up to 10 % are familial (fALS) 2 . The first gene associated with ALS (SOD1) 47 was described in 1993 4 . Since then, over 25 genes have been associated with fALS, sALS or both, 48 with the four most common being C9ORF72, SOD1, FUS, and TARDBP 2,5 . Notably, mutations in 49FUS are linked to both fALS and sALS, and present some of the earliest and more rapidly 50 progressive forms of the disease [6][7][8] . FUS is also involved in other neurodegenerative diseases, such 51 as frontotemporal dementia (FUS-FTLD), all characterized by FUS cytoplasmic inclusions 9 . 52 FUS is a nuclear RNA-binding protein, with a C-terminal nuclear localization signal (NLS) 9,10 . The 53 protein is involved in RNA processing, DNA repair and cellular proliferation, although its functions 54 are not precisely elucidated 9 . Most of the mutations associated with neurodegenerative diseases 55 alter FUS NLS [11][12][13] . More recently, mutations in FUS 3'UTR were described in ALS patients and 56 linked to an increased level of FUS mRNA and protein [14][15][16] . Surprisingly, over-expression of wild-57 type FUS provokes an aggressive ALS phenotype in mice and fruit flies, in accordance with findings 58 in yeast and mammalian cells [17][18][19][20][21] . The mechanism of the wild-type or ALS-linked mutated FUS 59 toxicity remains unclear 3,9,22 . 60With currently non-annotated proteins being increasingly reported 23-26 , we hypothesized that the 61 toxicity resulting from wild-type FUS over-expression may come from another, unseen, actor 26 . 62These novel proteins are coded by alternative open reading frames (altORFs) that are located 63 within "non-coding" RNAs (ncRNA), within the 5' or 3' "untranslated" regions (UTR) of mRNAs, or 64 overlapping a known coding sequence (CDS) within a different frame of an mRNA 25-27 . 65 Serendipitous discoveries and ribosome profiling have recently highlighted the distribution of 66 4 altORFs throughout the human genome, and the consequences of their absence from current 67 databases 26 . For example, mass spectrometry-based proteomics has become the gold standard 68 for protein identification and has been extensively used in ALS studies 28,29 . However, if a protein 69 is not annotated, it is not included in the protein database (e.g. UniProtKB), and thus cannot be 70 detected by mass spectrometry. An estimated 50 % of mass spectra from a proteomics 71 experiment are unmatched at the end of the analysis 26,30 . 72 Genome annotations must avoid spurious ORF annotations. Thus unless functional 73 characterization has been published, they rely upon 2 arbitrary criteria: a minimum length of 100 74 codons and a single ORF per transcript. Several groups developed tools to challenge such criteria, 75 such as the sORFs repository 31 and the OpenProt database 27 , which ...

show abstract

FUS pathology in ALS is linked to alterations in multiple ALS-associated proteins and rescued by drugs stimulating autophagy

Cited by 94 publications

References 33 publications

The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes

The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes

Reduced autophagy upon C9ORF72 loss synergizes with dipeptide repeat protein toxicity in G4C2 repeat expansion disorders

FUSgene is dual-coding with both proteins united in FUS-mediated toxicity

Contact Info

Product

Resources

About