Sil1-deficient fibroblasts generate an aberrant extracellular matrix leading to tendon disorganisation in Marinesco-Sjögren syndrome.

Cell attachment Cell motility Collagen Cytoskeleton DQ-collagen ER stress Electron micrograph Endoplasmic reticulum Extracellular matrix Skeletal muscle Tendon Transcriptomic analysis Unfolded protein response Woozy mouse Wound-healing

Journal

Journal of translational medicine
ISSN: 1479-5876
Titre abrégé: J Transl Med
Pays: England
ID NLM: 101190741

Informations de publication

Date de publication:
23 Aug 2024
Historique:
received: 29 05 2024
accepted: 05 08 2024
medline: 24 8 2024
pubmed: 24 8 2024
entrez: 23 8 2024
Statut: epublish

Résumé

Marinesco-Sjögren syndrome (MSS) is an autosomal recessive neuromuscular disorder that arises in early childhood and is characterized by congenital cataracts, myopathy associated with muscle weakness, and degeneration of Purkinje neurons leading to ataxia. About 60% of MSS patients have loss-of-function mutations in the SIL1 gene. Sil1 is an endoplasmic reticulum (ER) protein required for the release of ADP from the master chaperone Bip, which in turn will release the folded proteins. The expression of non-functional Sil1 leads to the accumulation of unfolded proteins in the ER and this triggers the unfolded protein response (UPR). A dysfunctional UPR could be a key element in the pathogenesis of MSS, although our knowledge of the molecular pathology of MSS is still incomplete. RNA-Seq transcriptomics was analysed using the String database and the Ingenuity Pathway Analysis platform. Fluorescence confocal microscopy was used to study the remodelling of the extracellular matrix (ECM). Transmission electron microscopy (TEM) was used to reveal the morphology of the ECM in vitro and in mouse tendon. Our transcriptomic analysis, performed on patient-derived fibroblasts, revealed 664 differentially expressed (DE) transcripts. Enrichment analysis of DE genes confirmed that the patient fibroblasts have a membrane trafficking issue. Furthermore, this analysis indicated that the extracellular space/ECM and the cell adhesion machinery, which together account for around 300 transcripts, could be affected in MSS. Functional assays showed that patient fibroblasts have a reduced capacity of ECM remodelling, reduced motility, and slower spreading during adhesion to Petri dishes. TEM micrographs of negative-stained ECM samples from these fibroblasts show differences of filaments in terms of morphology and size. Finally, structural analysis of the myotendinous junction of the soleus muscle and surrounding regions of the Achilles tendon revealed a disorganization of collagen fibres in the mouse model of MSS (woozy). ECM alterations can affect the proper functioning of several organs, including those damaged in MSS such as the central nervous system, skeletal muscle, bone and lens. On this basis, we propose that aberrant ECM is a key pathological feature of MSS and may help explain most of its clinical manifestations.

Sections du résumé

BACKGROUND BACKGROUND
Marinesco-Sjögren syndrome (MSS) is an autosomal recessive neuromuscular disorder that arises in early childhood and is characterized by congenital cataracts, myopathy associated with muscle weakness, and degeneration of Purkinje neurons leading to ataxia. About 60% of MSS patients have loss-of-function mutations in the SIL1 gene. Sil1 is an endoplasmic reticulum (ER) protein required for the release of ADP from the master chaperone Bip, which in turn will release the folded proteins. The expression of non-functional Sil1 leads to the accumulation of unfolded proteins in the ER and this triggers the unfolded protein response (UPR). A dysfunctional UPR could be a key element in the pathogenesis of MSS, although our knowledge of the molecular pathology of MSS is still incomplete.
METHODS METHODS
RNA-Seq transcriptomics was analysed using the String database and the Ingenuity Pathway Analysis platform. Fluorescence confocal microscopy was used to study the remodelling of the extracellular matrix (ECM). Transmission electron microscopy (TEM) was used to reveal the morphology of the ECM in vitro and in mouse tendon.
RESULTS RESULTS
Our transcriptomic analysis, performed on patient-derived fibroblasts, revealed 664 differentially expressed (DE) transcripts. Enrichment analysis of DE genes confirmed that the patient fibroblasts have a membrane trafficking issue. Furthermore, this analysis indicated that the extracellular space/ECM and the cell adhesion machinery, which together account for around 300 transcripts, could be affected in MSS. Functional assays showed that patient fibroblasts have a reduced capacity of ECM remodelling, reduced motility, and slower spreading during adhesion to Petri dishes. TEM micrographs of negative-stained ECM samples from these fibroblasts show differences of filaments in terms of morphology and size. Finally, structural analysis of the myotendinous junction of the soleus muscle and surrounding regions of the Achilles tendon revealed a disorganization of collagen fibres in the mouse model of MSS (woozy).
CONCLUSIONS CONCLUSIONS
ECM alterations can affect the proper functioning of several organs, including those damaged in MSS such as the central nervous system, skeletal muscle, bone and lens. On this basis, we propose that aberrant ECM is a key pathological feature of MSS and may help explain most of its clinical manifestations.

Identifiants

pubmed: 39180052
doi: 10.1186/s12967-024-05582-0
pii: 10.1186/s12967-024-05582-0
doi:

Substances chimiques

SIL1 protein, human 0
Guanine Nucleotide Exchange Factors 0
SIL1 protein, mouse 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

787

Subventions

Organisme : NextGenerationEU - MUR, Fondo Promozione e Sviluppo, DM 737/2021
ID : D75F21003210001
Organisme : Fondazione Telethon
ID : GGP20092
Organisme : Ministero dell'Università e della Ricerca
ID : DOT1353593-1

Informations de copyright

© 2024. The Author(s).

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Auteurs

Laura Amodei (L)

Department of Innovative Technologies in Medicine and Dentistry, Chieti, Italy.
Center for Advanced Studies and Technology (CAST), Chieti, Italy.

Anna Giulia Ruggieri (AG)

Department of Innovative Technologies in Medicine and Dentistry, Chieti, Italy.
Center for Advanced Studies and Technology (CAST), Chieti, Italy.

Francesca Potenza (F)

Department of Innovative Technologies in Medicine and Dentistry, Chieti, Italy.
Center for Advanced Studies and Technology (CAST), Chieti, Italy.

Marianna Viele (M)

Department of Innovative Technologies in Medicine and Dentistry, Chieti, Italy.
Center for Advanced Studies and Technology (CAST), Chieti, Italy.

Beatrice Dufrusine (B)

Department of Bioscience and Technology for Food Agriculture and Environment, University of Teramo, Teramo, 64100, Italy.

Raffaella Franciotti (R)

Department of Neuroscience, Imaging and Clinical Science, Chieti, Italy.

Laura Pietrangelo (L)

Department of Medicine and Aging Sciences, Chieti, Italy.

Matteo Ardini (M)

Department of Life, Health and Environmental Sciences, University of L'Aquila, L'Aquila, 67100, Italy.

Liborio Stuppia (L)

Center for Advanced Studies and Technology (CAST), Chieti, Italy.
Department of Psychological Health and Territorial Sciences, "G. d'Annunzio" University of Chieti-Pescara, Chieti, 66100, Italy.

Luca Federici (L)

Department of Innovative Technologies in Medicine and Dentistry, Chieti, Italy.
Center for Advanced Studies and Technology (CAST), Chieti, Italy.

Vincenzo De Laurenzi (V)

Department of Innovative Technologies in Medicine and Dentistry, Chieti, Italy.
Center for Advanced Studies and Technology (CAST), Chieti, Italy.

Michele Sallese (M)

Department of Innovative Technologies in Medicine and Dentistry, Chieti, Italy. Michele.sallese@unich.it.
Center for Advanced Studies and Technology (CAST), Chieti, Italy. Michele.sallese@unich.it.

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