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Gain-of-Function STIM1 L96V Mutation Causes Myogenesis Alteration in Muscle Cells From a Patient Affected by Tubular Aggregate Myopathy

Tubular Aggregate Myopathy (TAM) is a hereditary ultra-rare muscle disorder characterized by muscle weakness and cramps or myasthenic features. Biopsies from TAM patients show the presence of tubular aggregates originated from sarcoplasmic reticulum due to altered Ca(2+) homeostasis. TAM is caused b...

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Autores principales: Conte, Elena, Pannunzio, Alessandra, Imbrici, Paola, Camerino, Giulia Maria, Maggi, Lorenzo, Mora, Marina, Gibertini, Sara, Cappellari, Ornella, De Luca, Annamaria, Coluccia, Mauro, Liantonio, Antonella
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7952532/
https://www.ncbi.nlm.nih.gov/pubmed/33718371
http://dx.doi.org/10.3389/fcell.2021.635063
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author Conte, Elena
Pannunzio, Alessandra
Imbrici, Paola
Camerino, Giulia Maria
Maggi, Lorenzo
Mora, Marina
Gibertini, Sara
Cappellari, Ornella
De Luca, Annamaria
Coluccia, Mauro
Liantonio, Antonella
author_facet Conte, Elena
Pannunzio, Alessandra
Imbrici, Paola
Camerino, Giulia Maria
Maggi, Lorenzo
Mora, Marina
Gibertini, Sara
Cappellari, Ornella
De Luca, Annamaria
Coluccia, Mauro
Liantonio, Antonella
author_sort Conte, Elena
collection PubMed
description Tubular Aggregate Myopathy (TAM) is a hereditary ultra-rare muscle disorder characterized by muscle weakness and cramps or myasthenic features. Biopsies from TAM patients show the presence of tubular aggregates originated from sarcoplasmic reticulum due to altered Ca(2+) homeostasis. TAM is caused by gain-of-function mutations in STIM1 or ORAI1, proteins responsible for Store-Operated-Calcium-Entry (SOCE), a pivotal mechanism in Ca(2+) signaling. So far there is no cure for TAM and the mechanisms through which STIM1 or ORAI1 gene mutation lead to muscle dysfunction remain to be clarified. It has been established that post-natal myogenesis critically relies on Ca(2+) influx through SOCE. To explore how Ca(2+) homeostasis dysregulation associated with TAM impacts on muscle differentiation cascade, we here performed a functional characterization of myoblasts and myotubes deriving from patients carrying STIM1 L96V mutation by using fura-2 cytofluorimetry, high content imaging and real-time PCR. We demonstrated a higher resting Ca(2+) concentration and an increased SOCE in STIM1 mutant compared with control, together with a compensatory down-regulation of genes involved in Ca(2+) handling (RyR1, Atp2a1, Trpc1). Differentiating STIM1 L96V myoblasts persisted in a mononuclear state and the fewer multinucleated myotubes had distinct morphology and geometry of mitochondrial network compared to controls, indicating a defect in the late differentiation phase. The alteration in myogenic pathway was confirmed by gene expression analysis regarding early (Myf5, Mef2D) and late (DMD, Tnnt3) differentiation markers together with mitochondrial markers (IDH3A, OGDH). We provided evidences of mechanisms responsible for a defective myogenesis associated to TAM mutant and validated a reliable cellular model usefull for TAM preclinical studies.
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spelling pubmed-79525322021-03-13 Gain-of-Function STIM1 L96V Mutation Causes Myogenesis Alteration in Muscle Cells From a Patient Affected by Tubular Aggregate Myopathy Conte, Elena Pannunzio, Alessandra Imbrici, Paola Camerino, Giulia Maria Maggi, Lorenzo Mora, Marina Gibertini, Sara Cappellari, Ornella De Luca, Annamaria Coluccia, Mauro Liantonio, Antonella Front Cell Dev Biol Cell and Developmental Biology Tubular Aggregate Myopathy (TAM) is a hereditary ultra-rare muscle disorder characterized by muscle weakness and cramps or myasthenic features. Biopsies from TAM patients show the presence of tubular aggregates originated from sarcoplasmic reticulum due to altered Ca(2+) homeostasis. TAM is caused by gain-of-function mutations in STIM1 or ORAI1, proteins responsible for Store-Operated-Calcium-Entry (SOCE), a pivotal mechanism in Ca(2+) signaling. So far there is no cure for TAM and the mechanisms through which STIM1 or ORAI1 gene mutation lead to muscle dysfunction remain to be clarified. It has been established that post-natal myogenesis critically relies on Ca(2+) influx through SOCE. To explore how Ca(2+) homeostasis dysregulation associated with TAM impacts on muscle differentiation cascade, we here performed a functional characterization of myoblasts and myotubes deriving from patients carrying STIM1 L96V mutation by using fura-2 cytofluorimetry, high content imaging and real-time PCR. We demonstrated a higher resting Ca(2+) concentration and an increased SOCE in STIM1 mutant compared with control, together with a compensatory down-regulation of genes involved in Ca(2+) handling (RyR1, Atp2a1, Trpc1). Differentiating STIM1 L96V myoblasts persisted in a mononuclear state and the fewer multinucleated myotubes had distinct morphology and geometry of mitochondrial network compared to controls, indicating a defect in the late differentiation phase. The alteration in myogenic pathway was confirmed by gene expression analysis regarding early (Myf5, Mef2D) and late (DMD, Tnnt3) differentiation markers together with mitochondrial markers (IDH3A, OGDH). We provided evidences of mechanisms responsible for a defective myogenesis associated to TAM mutant and validated a reliable cellular model usefull for TAM preclinical studies. Frontiers Media S.A. 2021-02-26 /pmc/articles/PMC7952532/ /pubmed/33718371 http://dx.doi.org/10.3389/fcell.2021.635063 Text en Copyright © 2021 Conte, Pannunzio, Imbrici, Camerino, Maggi, Mora, Gibertini, Cappellari, De Luca, Coluccia and Liantonio. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Conte, Elena
Pannunzio, Alessandra
Imbrici, Paola
Camerino, Giulia Maria
Maggi, Lorenzo
Mora, Marina
Gibertini, Sara
Cappellari, Ornella
De Luca, Annamaria
Coluccia, Mauro
Liantonio, Antonella
Gain-of-Function STIM1 L96V Mutation Causes Myogenesis Alteration in Muscle Cells From a Patient Affected by Tubular Aggregate Myopathy
title Gain-of-Function STIM1 L96V Mutation Causes Myogenesis Alteration in Muscle Cells From a Patient Affected by Tubular Aggregate Myopathy
title_full Gain-of-Function STIM1 L96V Mutation Causes Myogenesis Alteration in Muscle Cells From a Patient Affected by Tubular Aggregate Myopathy
title_fullStr Gain-of-Function STIM1 L96V Mutation Causes Myogenesis Alteration in Muscle Cells From a Patient Affected by Tubular Aggregate Myopathy
title_full_unstemmed Gain-of-Function STIM1 L96V Mutation Causes Myogenesis Alteration in Muscle Cells From a Patient Affected by Tubular Aggregate Myopathy
title_short Gain-of-Function STIM1 L96V Mutation Causes Myogenesis Alteration in Muscle Cells From a Patient Affected by Tubular Aggregate Myopathy
title_sort gain-of-function stim1 l96v mutation causes myogenesis alteration in muscle cells from a patient affected by tubular aggregate myopathy
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7952532/
https://www.ncbi.nlm.nih.gov/pubmed/33718371
http://dx.doi.org/10.3389/fcell.2021.635063
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