RT Journal Article
SR Electronic
T1 Immortalized human myotonic dystrophy muscle cell lines to assess therapeutic compounds
JF Disease Models & Mechanisms
JO Dis Models Mech
FD The Company of Biologists Limited
SP 487
OP 497
DO 10.1242/dmm.027367
VO 10
IS 4
A1 Arandel, Ludovic
A1 Polay Espinoza, Micaela
A1 Matloka, Magdalena
A1 Bazinet, Audrey
A1 De Dea Diniz, Damily
A1 Naouar, Naïra
A1 Rau, Frédérique
A1 Jollet, Arnaud
A1 Edom-Vovard, Frédérique
A1 Mamchaoui, Kamel
A1 Tarnopolsky, Mark
A1 Puymirat, Jack
A1 Battail, Christophe
A1 Boland, Anne
A1 Deleuze, Jean-Francois
A1 Mouly, Vincent
A1 Klein, Arnaud F.
A1 Furling, Denis
YR 2017
UL http://dmm.biologists.org/content/10/4/487.abstract
AB Myotonic dystrophy type 1 (DM1) and type 2 (DM2) are autosomal dominant neuromuscular diseases caused by microsatellite expansions and belong to the family of RNA-dominant disorders. Availability of cellular models in which the DM mutation is expressed within its natural context is essential to facilitate efforts to identify new therapeutic compounds. Here, we generated immortalized DM1 and DM2 human muscle cell lines that display nuclear RNA aggregates of expanded repeats, a hallmark of myotonic dystrophy. Selected clones of DM1 and DM2 immortalized myoblasts behave as parental primary myoblasts with a reduced fusion capacity of immortalized DM1 myoblasts when compared with control and DM2 cells. Alternative splicing defects were observed in differentiated DM1 muscle cell lines, but not in DM2 lines. Splicing alterations did not result from differentiation delay because similar changes were found in immortalized DM1 transdifferentiated fibroblasts in which myogenic differentiation has been forced by overexpression of MYOD1. As a proof-of-concept, we show that antisense approaches alleviate disease-associated defects, and an RNA-seq analysis confirmed that the vast majority of mis-spliced events in immortalized DM1 muscle cells were affected by antisense treatment, with half of them significantly rescued in treated DM1 cells. Immortalized DM1 muscle cell lines displaying characteristic disease-associated molecular features such as nuclear RNA aggregates and splicing defects can be used as robust readouts for the screening of therapeutic compounds. Therefore, immortalized DM1 and DM2 muscle cell lines represent new models and tools to investigate molecular pathophysiological mechanisms and evaluate the in vitro effects of compounds on RNA toxicity associated with myotonic dystrophy mutations.