Distinct pathological signatures in human cellular models of myotonic dystrophy subtypes

Ellis Y. Kim, David Y. Barefield, Andy H. Vo, Anthony M. Gacita, Emma J. Schuster, Eugene Jeffrey Wyatt, Janel L. Davis, Biqin Dong, Cheng Sun, Patrick Page, Lisa Marie Castillo, Alexis R Demonbreun, Hao F Zhang, Elizabeth M McNally*

*Corresponding author for this work

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Myotonic dystrophy (DM) is the most common autosomal dominant muscular dystrophy and encompasses both skeletal muscle and cardiac complications. DM is nucleotide repeat expansion disorder in which type 1 (DM1) is due to a trinucleotide repeat expansion on chromosome 19 and type 2 (DM2) arises from a tetranucleotide repeat expansion on chromosome 3. Developing representative models of DM in animals has been challenging due to instability of nucleotide repeat expansions, especially for DM2, which is characterized by nucleotide repeat expansions often greater than 5,000 copies. To investigate mechanisms of human DM, we generated cellular models of DM1 and DM2. We used regulated MyoD expression to reprogram urine-derived cells into myotubes. In this myogenic cell model, we found impaired dystrophin expression, in the presence of muscleblind-like 1 (MBNL1) foci, and aberrant splicing in DM1 but not in DM2 cells. We generated induced pluripotent stem cells (iPSC) from healthy controls and DM1 and DM2 subjects, and we differentiated these into cardiomyocytes. DM1 and DM2 cells displayed an increase in RNA foci concomitant with cellular differentiation. iPSC-derived cardiomyocytes from DM1 but not DM2 had aberrant splicing of known target genes and MBNL sequestration. High-resolution imaging revealed tight association between MBNL clusters and RNA foci in DM1. Ca2+ transients differed between DM1- and DM2 iPSC–derived cardiomyocytes, and each differed from healthy control cells. RNA-sequencing from DM1- and DM2 iPSC–derived cardiomyocytes revealed distinct misregulation of gene expression, as well as differential aberrant splicing patterns. Together, these data support that DM1 and DM2, despite some shared clinical and molecular features, have distinct pathological signatures.

Original languageEnglish (US)
Article number122686
JournalJCI Insight
Volume4
Issue number6
DOIs
StatePublished - Mar 21 2019

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Myotonic Dystrophy
Cardiac Myocytes
Induced Pluripotent Stem Cells
Nucleotides
Trinucleotide Repeat Expansion
RNA
RNA Sequence Analysis
Chromosomes, Human, Pair 19
Dystrophin
Chromosomes, Human, Pair 3
Chromosomes, Human, Pair 2
Muscular Dystrophies
Skeletal Muscle Fibers
Microsatellite Repeats
Skeletal Muscle
Urine
Gene Expression
Genes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Kim, Ellis Y. ; Barefield, David Y. ; Vo, Andy H. ; Gacita, Anthony M. ; Schuster, Emma J. ; Wyatt, Eugene Jeffrey ; Davis, Janel L. ; Dong, Biqin ; Sun, Cheng ; Page, Patrick ; Castillo, Lisa Marie ; Demonbreun, Alexis R ; Zhang, Hao F ; McNally, Elizabeth M. / Distinct pathological signatures in human cellular models of myotonic dystrophy subtypes. In: JCI Insight. 2019 ; Vol. 4, No. 6.
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title = "Distinct pathological signatures in human cellular models of myotonic dystrophy subtypes",
abstract = "Myotonic dystrophy (DM) is the most common autosomal dominant muscular dystrophy and encompasses both skeletal muscle and cardiac complications. DM is nucleotide repeat expansion disorder in which type 1 (DM1) is due to a trinucleotide repeat expansion on chromosome 19 and type 2 (DM2) arises from a tetranucleotide repeat expansion on chromosome 3. Developing representative models of DM in animals has been challenging due to instability of nucleotide repeat expansions, especially for DM2, which is characterized by nucleotide repeat expansions often greater than 5,000 copies. To investigate mechanisms of human DM, we generated cellular models of DM1 and DM2. We used regulated MyoD expression to reprogram urine-derived cells into myotubes. In this myogenic cell model, we found impaired dystrophin expression, in the presence of muscleblind-like 1 (MBNL1) foci, and aberrant splicing in DM1 but not in DM2 cells. We generated induced pluripotent stem cells (iPSC) from healthy controls and DM1 and DM2 subjects, and we differentiated these into cardiomyocytes. DM1 and DM2 cells displayed an increase in RNA foci concomitant with cellular differentiation. iPSC-derived cardiomyocytes from DM1 but not DM2 had aberrant splicing of known target genes and MBNL sequestration. High-resolution imaging revealed tight association between MBNL clusters and RNA foci in DM1. Ca2+ transients differed between DM1- and DM2 iPSC–derived cardiomyocytes, and each differed from healthy control cells. RNA-sequencing from DM1- and DM2 iPSC–derived cardiomyocytes revealed distinct misregulation of gene expression, as well as differential aberrant splicing patterns. Together, these data support that DM1 and DM2, despite some shared clinical and molecular features, have distinct pathological signatures.",
author = "Kim, {Ellis Y.} and Barefield, {David Y.} and Vo, {Andy H.} and Gacita, {Anthony M.} and Schuster, {Emma J.} and Wyatt, {Eugene Jeffrey} and Davis, {Janel L.} and Biqin Dong and Cheng Sun and Patrick Page and Castillo, {Lisa Marie} and Demonbreun, {Alexis R} and Zhang, {Hao F} and McNally, {Elizabeth M}",
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Distinct pathological signatures in human cellular models of myotonic dystrophy subtypes. / Kim, Ellis Y.; Barefield, David Y.; Vo, Andy H.; Gacita, Anthony M.; Schuster, Emma J.; Wyatt, Eugene Jeffrey; Davis, Janel L.; Dong, Biqin; Sun, Cheng; Page, Patrick; Castillo, Lisa Marie; Demonbreun, Alexis R; Zhang, Hao F; McNally, Elizabeth M.

In: JCI Insight, Vol. 4, No. 6, 122686, 21.03.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Distinct pathological signatures in human cellular models of myotonic dystrophy subtypes

AU - Kim, Ellis Y.

AU - Barefield, David Y.

AU - Vo, Andy H.

AU - Gacita, Anthony M.

AU - Schuster, Emma J.

AU - Wyatt, Eugene Jeffrey

AU - Davis, Janel L.

AU - Dong, Biqin

AU - Sun, Cheng

AU - Page, Patrick

AU - Castillo, Lisa Marie

AU - Demonbreun, Alexis R

AU - Zhang, Hao F

AU - McNally, Elizabeth M

PY - 2019/3/21

Y1 - 2019/3/21

N2 - Myotonic dystrophy (DM) is the most common autosomal dominant muscular dystrophy and encompasses both skeletal muscle and cardiac complications. DM is nucleotide repeat expansion disorder in which type 1 (DM1) is due to a trinucleotide repeat expansion on chromosome 19 and type 2 (DM2) arises from a tetranucleotide repeat expansion on chromosome 3. Developing representative models of DM in animals has been challenging due to instability of nucleotide repeat expansions, especially for DM2, which is characterized by nucleotide repeat expansions often greater than 5,000 copies. To investigate mechanisms of human DM, we generated cellular models of DM1 and DM2. We used regulated MyoD expression to reprogram urine-derived cells into myotubes. In this myogenic cell model, we found impaired dystrophin expression, in the presence of muscleblind-like 1 (MBNL1) foci, and aberrant splicing in DM1 but not in DM2 cells. We generated induced pluripotent stem cells (iPSC) from healthy controls and DM1 and DM2 subjects, and we differentiated these into cardiomyocytes. DM1 and DM2 cells displayed an increase in RNA foci concomitant with cellular differentiation. iPSC-derived cardiomyocytes from DM1 but not DM2 had aberrant splicing of known target genes and MBNL sequestration. High-resolution imaging revealed tight association between MBNL clusters and RNA foci in DM1. Ca2+ transients differed between DM1- and DM2 iPSC–derived cardiomyocytes, and each differed from healthy control cells. RNA-sequencing from DM1- and DM2 iPSC–derived cardiomyocytes revealed distinct misregulation of gene expression, as well as differential aberrant splicing patterns. Together, these data support that DM1 and DM2, despite some shared clinical and molecular features, have distinct pathological signatures.

AB - Myotonic dystrophy (DM) is the most common autosomal dominant muscular dystrophy and encompasses both skeletal muscle and cardiac complications. DM is nucleotide repeat expansion disorder in which type 1 (DM1) is due to a trinucleotide repeat expansion on chromosome 19 and type 2 (DM2) arises from a tetranucleotide repeat expansion on chromosome 3. Developing representative models of DM in animals has been challenging due to instability of nucleotide repeat expansions, especially for DM2, which is characterized by nucleotide repeat expansions often greater than 5,000 copies. To investigate mechanisms of human DM, we generated cellular models of DM1 and DM2. We used regulated MyoD expression to reprogram urine-derived cells into myotubes. In this myogenic cell model, we found impaired dystrophin expression, in the presence of muscleblind-like 1 (MBNL1) foci, and aberrant splicing in DM1 but not in DM2 cells. We generated induced pluripotent stem cells (iPSC) from healthy controls and DM1 and DM2 subjects, and we differentiated these into cardiomyocytes. DM1 and DM2 cells displayed an increase in RNA foci concomitant with cellular differentiation. iPSC-derived cardiomyocytes from DM1 but not DM2 had aberrant splicing of known target genes and MBNL sequestration. High-resolution imaging revealed tight association between MBNL clusters and RNA foci in DM1. Ca2+ transients differed between DM1- and DM2 iPSC–derived cardiomyocytes, and each differed from healthy control cells. RNA-sequencing from DM1- and DM2 iPSC–derived cardiomyocytes revealed distinct misregulation of gene expression, as well as differential aberrant splicing patterns. Together, these data support that DM1 and DM2, despite some shared clinical and molecular features, have distinct pathological signatures.

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