FUS ALS neurons activate major stress pathways and reduce translation as an early protective mechanism against neurodegeneration

Szewczyk, Barbara; Günther, René; Japtok, Julia; Frech, Moritz J.; Naumann, Marcel; Hermann, Andreas

doi:10.1016/j.celrep.2023.112025

Cited by 18 publications

(20 citation statements)

References 82 publications

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“…The protein embeds a IDR which is responsible for self-assembly and spontaneous aggregation [24][25][26] ; ALS-linked mutations are clustered in the Nuclear Localization Sequence (NLS) reducing its nuclear reimport and resulting in an aberrant cytoplasmic accumulation and subsequent recruitment in SG 27,28 . As mutant FUS accumulates in the cytoplasm of motor-neurons, it triggers the activation of major stress pathways and impacts SG number and dynamics, finally leading to an increased vulnerability of these cells to harmful stimuli 29 . In this work, we focused on the RNA content of SG in a neuronal-like system, demonstrating that neuronal RNAs are specifically enriched in these condensates in response to oxidative stress.…”

Section: Introductionmentioning

confidence: 99%

ALS-associated FUS mutation reshapes the RNA and protein composition of Stress Granules

Mariani,

Setti,

Castagnetti

et al. 2023

Preprint

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Stress Granules (SG) formation is a cellular protection mechanism in harmful conditions, constituting a storage for untranslated mRNAs and RNA-binding proteins (RBPs); however, these condensates can turn into pathological aggregates, that in neurons are related to the onset of neurodegenerative diseases, like Amyotrophic Lateral Sclerosis (ALS). Mutations in the RBP FUS leading to its cytoplasmic mis-localization are causatively linked to ALS, since they mediate its accumulation in SGs, triggering their transition towards cytotoxic inclusions. Here, we describe the SG transcriptome in a neural context and compare with datasets from other systems, identifying both common rules and diversifying characteristics for SG recruitment of transcripts. We demonstrate that SG dynamics and RNA content are strongly modified by the incorporation of aberrantly localized mutant FUS, switching to a more unstructured, AU-rich SG transcriptome and losing a wide population of GC-rich, structured RNAs. We show that these alterations mainly depend on mutant FUS which favors the SG incorporation of its protein interactors and in turn of their target RNAs. Our data give a comprehensive view of the molecular differences between physiological and pathological SG in ALS conditions, showing how a single mutation is sufficient to impact the RNA and protein population of these condensates.

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Section: Introductionmentioning

confidence: 99%

ALS-associated FUS mutation reshapes the RNA and protein composition of Stress Granules

Mariani,

Setti,

Castagnetti

et al. 2023

Preprint

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“…ALS-causing mutations in FUS are mainly localized in its NLS and thus causing a cytoplasmic mislocalization, i.e. aggregation (Japtok et al, 2015, Szewczyk et al, 2023, Szewczyk et al, 2021). This is accompanied by a loss of nuclear FUS function including a lack of proper recruitment of FUS to DNA damage sites and DNA damage repair (DDR) (Naumann et al, 2018, Wang et al, 2018).…”

Section: Resultsmentioning

confidence: 99%

“…Most importantly, restoration of mitochondrial inner membrane potential delayed neurodegeneration in FUS-ALS (Naumann et al, 2018). ALS-causing mutations in FUS are mainly localized in its nuclear localization sequence (NLS) and thus causing a cytoplasmic mislocalization (Japtok, Lojewski et al, 2015, Szewczyk, Gunther et al, 2023, Szewczyk, Gunther et al, 2021. This is accompanied by a loss of nuclear FUS function including a lack of proper recruitment of FUS to DNA damage sites and DNA damage repair (DDR) (Naumann et al, 2018, Wang, Guo et al, 2018, a mechanism downstream of poly(ADPribose) polymerase 1 (PARP1) (Mastrocola, Kim et al, 2013, Rulten, Rotheray et al, 2014.…”

Section: Introductionmentioning

confidence: 99%

DJ-1 products glycolic acid and D-lactate restore deficient axonal trafficking and DNA damage response in FUS and SOD1-associated amyotrophic lateral sclerosis

Pal,

Grossmann,

Glaß

et al. 2023

Preprint

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Amyotrophic Lateral Sclerosis (ALS) is the most common motor neuron disease leading to death within 2-5 years. Currently available drugs can only slightly prolong survival. Despite the progress that has been achieved in unravelling the molecular mechanisms of the disease so far, the underlying pathophysiology is not fully understood. We present novel insights into the pathophysiology of Superoxide Dismutase 1 (SOD1)- and in particular Fused In Sarcoma (FUS)-ALS by revealing a putatively central role of the Parkinsons disease (PD) associated glyoxylase DJ-1 and its products glycolic acid (GA) and D-lactic acid (DL). Combined, but not single, treatment with GA and DL restored axonal trafficking deficits of mitochondria and lysosomes in FUS- and SOD1-ALS patient-derived motoneurons (MNs). This was accompanied by restoration of mitochondrial membrane potential as well as mitochondrial fragmentation (FUS-ALS) or elongation (SOD1-ALS). Furthermore, GA and DL restored cytoplasmic mislocalization of FUS and FUS recruitment to DNA damage sites. We further show that despite presenting an early axonal transport deficiency as well, TDP-43 patient-derived MNs did not share this mechanism. While this points towards the necessity of individualized (gene-) specific therapy stratification, it also suggests common therapeutic targets across different gene variants of ALS. Thus, we introduce a putative novel treatment for ALS based on a combination of the two substances GA and DL which might be not only an interesting novel drug candidate in subsets of ALS cases but also in other neurodegenerative diseases characterized by mitochondrial depolarization.

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“…Induced pluripotent stem cells (iPSCs)-derived small molecule neuroprogenitor cells (smNPCs), produced as described in 57,75 , were differentiated to reach 2-weeks matured motor neurons. The iPSC generation and respective characterizations was previously reported 57 and approved by the local ethics committee (EK45022009).…”

Section: Author Contributionsmentioning

confidence: 99%

Small molecule inhibitors of hnRNPA2B1-RNA interactions reveal a predictable sorting of RNA subsets into extracellular vesicles

Corsi,

Peroni,

Belli

et al. 2024

Preprint

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Extracellular vesicles (EVs) are cell-secreted membranous particles contributing to intercellular communication. Coding and non-coding RNAs are widely detected EV cargo, and RNA-binding proteins (RBPs), such as hnRNPA2B1, have been circumstantially implicated in sorting vesicular RNAs. However, the contribution of competitive RBP-RNA interactions responsible for RNA-sorting outcomes still needs to be deciphered, especially for EV-RNA interference and predictability. We conducted a reverse proteomic analysis that prioritized heterogeneous nuclear ribonucleoproteins recognizing purine-rich RNA sequences representing a subset of previously identified EXO motifs. A screening campaign using a full-length human hnRNPA2B1 protein and artificial purine-rich RNA brought to small molecule inhibitors orthogonally validated through biochemical and cell-based approaches. Selected drugs effectively interfered with a post-transcriptional layer impacting secreted EV- RNAs, reducing the vesicular pro-inflammatory miR-221 while counteracting the hnRNPA2B1- or TDP43Q331K-dependent paracrine activation of NF-κB in EV-recipient cells. This study demonstrates the possibility of predicting the EV-RNA quality for developing innovative strategies targeting discrete paracrine functions.SummaryExtracellular vesicles (EVs) are cell-released, heterogeneous lipid particles conceived as vehicles for intercellular communication. RNA is a widely detected cargo, and the comprehension of EV-sorting mechanisms represents a step forward in predicting EV quality and associated paracrine effects. While it is known that specific RNA-binding proteins (RBPs) play a role in EV-RNA sorting, the quantitative contribution of competing RBP-RNA interactions and the predictability of RNA-sorting outcomes are poorly understood. Here, we show that a core of hnRNPs compete for the binding to the heterogeneous EV-RNAin vitro. Given prioritized interactions with purine-rich RNA motifs, we set up a pharmacological screen platform to find inhibitors of protein-RNA interactions. Our results suggest that selected small molecules can interfere with EV-RNA quality, altering the distribution of specific miRNA cargoes and associating with a discriminant NF-kB activation in EV-recipient cells. This work highlights the role of RBP-RNA interactions in influencing the EV-RNA quality and paracrine functions.

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FUS ALS neurons activate major stress pathways and reduce translation as an early protective mechanism against neurodegeneration

Cited by 18 publications

References 82 publications

ALS-associated FUS mutation reshapes the RNA and protein composition of Stress Granules

ALS-associated FUS mutation reshapes the RNA and protein composition of Stress Granules

DJ-1 products glycolic acid and D-lactate restore deficient axonal trafficking and DNA damage response in FUS and SOD1-associated amyotrophic lateral sclerosis

Small molecule inhibitors of hnRNPA2B1-RNA interactions reveal a predictable sorting of RNA subsets into extracellular vesicles

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