Immortalized Human Myotonic Dystrophy Type 1 Muscle Cell Lines to Address Patient Heterogeneity

Overview

Journal iScience

Publisher Cell Press

Date 2024 Jun 4

PMID 38832025

Authors

Judit Nunez-Manchon

Julia Capo

Alicia Martinez-Pineiro

Eduard Juanola

Jovan Pesovic

Laura Mosqueira-Martin

Klaudia Gonzalez-Imaz

Pau Maestre-Mora

Renato Odria

Estefania Cerro-Herreros

Neia Naldaiz-Gastesi

Adolfo Lopez de Munain

Ruben Artero

Dusanka Savic-Pavicevic

Ainara Vallejo-Illarramendi

Kamel Mamchaoui

Anne Bigot

Vincent Mouly

Monica Suelves

Gisela Nogales-Gadea

Affiliations

Soon will be listed here.

Abstract

Historically, cellular models have been used as a tool to study myotonic dystrophy type 1 (DM1) and the validation of therapies in said pathology. However, there is a need for models that represent the clinical heterogeneity observed in patients with DM1 that is lacking in classical models. In this study, we immortalized three DM1 muscle lines derived from patients with different DM1 subtypes and clinical backgrounds and characterized them at the genetic, epigenetic, and molecular levels. All three cell lines display DM1 hallmarks, such as the accumulation of RNA foci, MBNL1 sequestration, splicing alterations, and reduced fusion. In addition, alterations in early myogenic markers, myotube diameter and CTCF1 DNA methylation were also found in DM1 cells. Notably, the new lines show a high level of heterogeneity in both the size of the CTG expansion and the aforementioned molecular alterations. Importantly, these immortalized cells also responded to previously tested therapeutics. Altogether, our results show that these three human DM1 cellular models are suitable to study the pathophysiological heterogeneity of DM1 and to test future therapeutic options.

References

Arandel L, Polay Espinoza M, Matloka M, Bazinet A, De Dea Diniz D, Naouar N . Immortalized human myotonic dystrophy muscle cell lines to assess therapeutic compounds. Dis Model Mech. 2017; 10(4):487-497. PMC: 5399563. DOI: 10.1242/dmm.027367. View

Martorell L, Martinez J, Carey N, Johnson K, Baiget M . Comparison of CTG repeat length expansion and clinical progression of myotonic dystrophy over a five year period. J Med Genet. 1995; 32(8):593-6. PMC: 1051631. DOI: 10.1136/jmg.32.8.593. View

Yamashita Y, Matsuura T, Kurosaki T, Amakusa Y, Kinoshita M, Ibi T . LDB3 splicing abnormalities are specific to skeletal muscles of patients with myotonic dystrophy type 1 and alter its PKC binding affinity. Neurobiol Dis. 2014; 69:200-5. DOI: 10.1016/j.nbd.2014.05.026. View

Klesert T, Cho D, Clark J, Maylie J, Adelman J, Snider L . Mice deficient in Six5 develop cataracts: implications for myotonic dystrophy. Nat Genet. 2000; 25(1):105-9. DOI: 10.1038/75490. View

Kimura T, Nakamori M, Lueck J, Pouliquin P, Aoike F, Fujimura H . Altered mRNA splicing of the skeletal muscle ryanodine receptor and sarcoplasmic/endoplasmic reticulum Ca2+-ATPase in myotonic dystrophy type 1. Hum Mol Genet. 2005; 14(15):2189-200. DOI: 10.1093/hmg/ddi223. View

Miller J, Urbinati C, Stenberg M, Byrne B, Thornton C, Swanson M . Recruitment of human muscleblind proteins to (CUG)(n) expansions associated with myotonic dystrophy. EMBO J. 2000; 19(17):4439-48. PMC: 302046. DOI: 10.1093/emboj/19.17.4439. View

Antequera F, Boyes J, Bird A . High levels of de novo methylation and altered chromatin structure at CpG islands in cell lines. Cell. 1990; 62(3):503-14. DOI: 10.1016/0092-8674(90)90015-7. View

Rasmussen A, Hildonen M, Vissing J, Duno M, Tumer Z, Birkedal U . High Resolution Analysis of Hypermethylation and Repeat Interruptions in Myotonic Dystrophy Type 1. Genes (Basel). 2022; 13(6). PMC: 9222588. DOI: 10.3390/genes13060970. View

Tsai Y, de Pontual L, Heiner C, Stojkovic T, Furling D, Bassez G . Identification of a CCG-Enriched Expanded Allele in Patients with Myotonic Dystrophy Type 1 Using Amplification-Free Long-Read Sequencing. J Mol Diagn. 2022; 24(11):1143-1154. DOI: 10.1016/j.jmoldx.2022.08.003. View

10.

Pantic B, Borgia D, Giunco S, Malena A, Kiyono T, Salvatori S . Reliable and versatile immortal muscle cell models from healthy and myotonic dystrophy type 1 primary human myoblasts. Exp Cell Res. 2016; 342(1):39-51. DOI: 10.1016/j.yexcr.2016.02.013. View

11.

Westerlaken J, Van der Zee C, Peters W, Wieringa B . The DMWD protein from the myotonic dystrophy (DM1) gene region is developmentally regulated and is present most prominently in synapse-dense brain areas. Brain Res. 2003; 971(1):116-27. DOI: 10.1016/s0006-8993(03)02430-2. View

12.

Jones P, Wolkowicz M, Rideout 3rd W, Gonzales F, Marziasz C, Coetzee G . De novo methylation of the MyoD1 CpG island during the establishment of immortal cell lines. Proc Natl Acad Sci U S A. 1990; 87(16):6117-21. PMC: 54483. DOI: 10.1073/pnas.87.16.6117. View

13.

Yamamoto T, Miura A, Itoh K, Takeshima Y, Nishio H . RNA sequencing reveals abnormal LDB3 splicing in sudden cardiac death. Forensic Sci Int. 2019; 302:109906. DOI: 10.1016/j.forsciint.2019.109906. View

14.

Wojtkowiak-Szlachcic A, Taylor K, Stepniak-Konieczna E, Sznajder L, Mykowska A, Sroka J . Short antisense-locked nucleic acids (all-LNAs) correct alternative splicing abnormalities in myotonic dystrophy. Nucleic Acids Res. 2015; 43(6):3318-31. PMC: 4381072. DOI: 10.1093/nar/gkv163. View

15.

Spitalieri P, Talarico R, Caioli S, Murdocca M, Serafino A, Girasole M . Modelling the pathogenesis of Myotonic Dystrophy type 1 cardiac phenotype through human iPSC-derived cardiomyocytes. J Mol Cell Cardiol. 2018; 118:95-109. DOI: 10.1016/j.yjmcc.2018.03.012. View

16.

Savkur R, Philips A, Cooper T . Aberrant regulation of insulin receptor alternative splicing is associated with insulin resistance in myotonic dystrophy. Nat Genet. 2001; 29(1):40-7. DOI: 10.1038/ng704. View

17.

Gomes-Pereira M, Bidichandani S, Monckton D . Analysis of unstable triplet repeats using small-pool polymerase chain reaction. Methods Mol Biol. 2004; 277:61-76. DOI: 10.1385/1-59259-804-8:061. View

18.

Pesovic J, Peric S, Brkusanin M, Brajuskovic G, Rakocevic-Stojanovic V, Savic-Pavicevic D . Repeat Interruptions Modify Age at Onset in Myotonic Dystrophy Type 1 by Stabilizing Expansions in Somatic Cells. Front Genet. 2018; 9:601. PMC: 6278776. DOI: 10.3389/fgene.2018.00601. View

19.

Cumming S, Jimenez-Moreno C, Okkersen K, Wenninger S, Daidj F, Hogarth F . Genetic determinants of disease severity in the myotonic dystrophy type 1 OPTIMISTIC cohort. Neurology. 2019; 93(10):e995-e1009. PMC: 6745735. DOI: 10.1212/WNL.0000000000008056. View

20.

Mamchaoui K, Trollet C, Bigot A, Negroni E, Chaouch S, Wolff A . Immortalized pathological human myoblasts: towards a universal tool for the study of neuromuscular disorders. Skelet Muscle. 2011; 1:34. PMC: 3235972. DOI: 10.1186/2044-5040-1-34. View