Molecular characterization of myotonic dystrophy fibroblast cell lines for use in small molecule screening
Jana R. Jenquin,
Alana P. O’Brien,
Kiril Poukalov,
Yidan Lu,
Jesus A. Frias,
Hannah K. Shorrock,
Jared I. Richardson,
Hormoz Mazdiyasni,
Hongfen Yang,
Robert W. Huigens, III,
David Boykin,
Laura P.W. Ranum,
John Douglas Cleary,
Eric T. Wang,
J. Andrew Berglund
Affiliations
Jana R. Jenquin
Department of Biochemistry and Molecular Biology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA; RNA Institute, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA
Alana P. O’Brien
Department of Molecular Genetics and Microbiology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA
Kiril Poukalov
Department of Molecular Genetics and Microbiology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA
Yidan Lu
Department of Molecular Genetics and Microbiology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA
Jesus A. Frias
RNA Institute, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA; Department of Biological Sciences, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA
Hannah K. Shorrock
RNA Institute, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA
Jared I. Richardson
Department of Biochemistry and Molecular Biology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA; RNA Institute, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA; Department of Biological Sciences, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA
Hormoz Mazdiyasni
RNA Institute, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA
Hongfen Yang
Department of Medicinal Chemistry, Center for Natural Products Drug Discovery and Development, College of Pharmacy, University of Florida, Gainesville, FL 32610, USA
Robert W. Huigens, III
Department of Medicinal Chemistry, Center for Natural Products Drug Discovery and Development, College of Pharmacy, University of Florida, Gainesville, FL 32610, USA
David Boykin
Department of Chemistry, Georgia State University, Atlanta, GA 30303, USA
Laura P.W. Ranum
Department of Molecular Genetics and Microbiology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA
John Douglas Cleary
RNA Institute, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA
Eric T. Wang
Department of Molecular Genetics and Microbiology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA
J. Andrew Berglund
Department of Biochemistry and Molecular Biology, Center for NeuroGenetics, College of Medicine, University of Florida, Gainesville, FL 32610, USA; RNA Institute, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA; Department of Biological Sciences, College of Arts and Sciences, University at Albany-SUNY, Albany, NY 12222, USA; Corresponding author
Summary: Myotonic dystrophy type 1 (DM1) and type 2 (DM2) are common forms of adult onset muscular dystrophy. Pathogenesis in both diseases is largely driven by production of toxic-expanded repeat RNAs that sequester MBNL RNA-binding proteins, causing mis-splicing. Given this shared pathogenesis, we hypothesized that diamidines, small molecules that rescue mis-splicing in DM1 models, could also rescue mis-splicing in DM2 models. While several DM1 cell models exist, few are available for DM2 limiting research and therapeutic development. Here, we characterize DM1 and DM2 patient-derived fibroblasts for use in small molecule screens and therapeutic studies. We identify mis-splicing events unique to DM2 fibroblasts and common events shared with DM1 fibroblasts. We show that diamidines can partially rescue molecular phenotypes in both DM1 and DM2 fibroblasts. This study demonstrates the potential of fibroblasts as models for DM1 and DM2, which will help meet an important need for well-characterized DM2 cell models.
