Skip to Content
Merck
  • HDAC inhibitors tune miRNAs in extracellular vesicles of dystrophic muscle-resident mesenchymal cells.

HDAC inhibitors tune miRNAs in extracellular vesicles of dystrophic muscle-resident mesenchymal cells.

EMBO reports (2020-08-06)
Martina Sandonà, Silvia Consalvi, Luca Tucciarone, Marco De Bardi, Manuel Scimeca, Daniela Francesca Angelini, Valentina Buffa, Adele D'Amico, Enrico Silvio Bertini, Sara Cazzaniga, Paolo Bettica, Marina Bouché, Antonella Bongiovanni, Pier Lorenzo Puri, Valentina Saccone
ABSTRACT

We show that extracellular vesicles (EVs) released by mesenchymal cells (i.e., fibro-adipogenic progenitors-FAPs) mediate microRNA (miR) transfer to muscle stem cells (MuSCs) and that exposure of dystrophic FAPs to HDAC inhibitors (HDACis) increases the intra-EV levels of a subset of miRs, which cooperatively target biological processes of therapeutic interest, including regeneration, fibrosis, and inflammation. Increased levels of miR-206 in EVs released by FAPs of muscles from Duchenne muscular dystrophy (DMD) patients or mdx mice exposed to HDACi are associated with enhanced regeneration and decreased fibrosis. Consistently, EVs from HDACi-treated dystrophic FAPs can stimulate MuSC activation and expansion ex vivo, and promote regeneration, while inhibiting fibrosis and inflammation of dystrophic muscles, upon intramuscular transplantation in mdx mice, in vivo. AntagomiR-mediated blockade of individual miRs reveals a specific requirement of miR-206 for EV-induced expansion of MuSCs and regeneration of dystrophic muscles, and indicates that cooperative activity of HDACi-induced miRs accounts for the net biological effect of these EVs. These data point to pharmacological modulation of EV content as novel strategy for therapeutic interventions in muscular dystrophies.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
GW4869, ≥90% (NMR)
Sigma-Aldrich
Anti-Laminin antibody produced in rabbit, 0.5 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Sodium chloride, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Formamide, BioReagent, ≥99.5% (GC), for molecular biology
Sigma-Aldrich
Acridine Orange hydrochloride solution, ≥95.0% (HPLC), 10 mg/mL in H2O
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, fatty acid free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
Trizma® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
Sigma-Aldrich
TWEEN® 20, viscous liquid
Sigma-Aldrich
Glycine, suitable for electrophoresis, ≥99%
Roche
Anti-Digoxigenin-AP, Fab fragments, from sheep
Sigma-Aldrich
Acetic acid, glacial, puriss., 99-100%
Sigma-Aldrich
Methanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
Sigma-Aldrich
Acetone, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)
Sigma-Aldrich
Bouin′s solution, histological fixative
Sigma-Aldrich
Acridine Orange base, Dye content 75 %
Sigma-Aldrich
Deoxyribonucleic acid, single stranded from salmon testes, For hybridization
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
(−)-Tetramisole hydrochloride, ≥99% (GC)
Sigma-Aldrich
Biebrich Scarlet-Acid Fuchsin Solution
Sigma-Aldrich
TRI Reagent®, For processing tissues, cells cultured in monolayer or cell pellets
Sigma-Aldrich
Weigert′s Iron Hematoxylin Set, Set
Sigma-Aldrich
Ribonucleic acid, transfer from Escherichia coli, Type XX, Strain W, lyophilized powder
Sigma-Aldrich
Dextran sulfate sodium salt from Leuconostoc spp., for molecular biology, average Mw >500,000 (dextran starting material), contains 0.5-2% phosphate buffer
Sigma-Aldrich
Trichostatin A, ≥98% (HPLC), from Streptomyces sp.
Sigma-Aldrich
o-Xylene, reagent grade, ≥98.0%