iPSC Modeling of RBM20-Deficient DCM Identifies Upregulation of RBM20 as a Therapeutic Strategy

Francesca Briganti; Han Sun; Wu Wei; Jingyan Wu; Chenchen Zhu; Martin Liss; Ioannis Karakikes; Shannon Rego; Andrea Cipriano; Michael Snyder; Benjamin Meder; Zhenyu Xu; Gilles Millat; Michael Gotthardt; Mark Mercola; Lars M Steinmetz

doi:10.1016/j.celrep.2020.108117

iPSC Modeling of RBM20-Deficient DCM Identifies Upregulation of RBM20 as a Therapeutic Strategy

Cell Rep. 2020 Sep 8;32(10):108117. doi: 10.1016/j.celrep.2020.108117.

Authors

Francesca Briganti¹, Han Sun², Wu Wei³, Jingyan Wu², Chenchen Zhu², Martin Liss⁴, Ioannis Karakikes⁵, Shannon Rego², Andrea Cipriano⁶, Michael Snyder², Benjamin Meder⁷, Zhenyu Xu⁸, Gilles Millat⁹, Michael Gotthardt¹⁰, Mark Mercola¹¹, Lars M Steinmetz¹²

Affiliations

¹ European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Heidelberg, Germany; Collaboration for joint PhD degree between EMBL and Heidelberg University, Faculty of Biosciences, Heidelberg, Germany; Department of Genetics, School of Medicine, Stanford University, Stanford, CA, USA; Cardiovascular Institute and Department of Medicine, Stanford University, Stanford, CA, USA.
² Department of Genetics, School of Medicine, Stanford University, Stanford, CA, USA.
³ Stanford Genome Technology Center, Stanford University, Palo Alto, CA, USA.
⁴ Neuromuscular and Cardiovascular Cell Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany.
⁵ Cardiovascular Institute and Department of Cardiothoracic Surgery, Stanford University, Stanford, CA, USA.
⁶ Department of Obstetrics and Gynecology, Stanford University, Stanford, CA, USA.
⁷ Stanford Genome Technology Center, Stanford University, Palo Alto, CA, USA; Institute for Cardiomyopathies Heidelberg and Department of Internal Medicine III, University of Heidelberg, Heidelberg, Germany.
⁸ SOPHiA Genetics, St. Sulpice, Switzerland.
⁹ Laboratoire de Cardiogénétique Moléculaire, Centre de Biologie et Pathologie Est, Hospices Civils de Lyon, Lyon, France.
¹⁰ Neuromuscular and Cardiovascular Cell Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany; Department of Cardiology, Charité-Universitätsmedizin Berlin, Berlin, Germany; DZHK: German Center for Cardiovascular Research, Partner Site Berlin, Berlin, Germany.
¹¹ Cardiovascular Institute and Department of Medicine, Stanford University, Stanford, CA, USA. Electronic address: mmercola@stanford.edu.
¹² European Molecular Biology Laboratory (EMBL), Genome Biology Unit, Heidelberg, Germany; Department of Genetics, School of Medicine, Stanford University, Stanford, CA, USA; Cardiovascular Institute and Department of Medicine, Stanford University, Stanford, CA, USA; Stanford Genome Technology Center, Stanford University, Palo Alto, CA, USA; DZHK: German Center for Cardiovascular Research, Partner Site EMBL Heidelberg, Heidelberg, Germany. Electronic address: larsms@stanford.edu.

Abstract

Recent advances in induced pluripotent stem cell (iPSC) technology and directed differentiation of iPSCs into cardiomyocytes (iPSC-CMs) make it possible to model genetic heart disease in vitro. We apply CRISPR/Cas9 genome editing technology to introduce three RBM20 mutations in iPSCs and differentiate them into iPSC-CMs to establish an in vitro model of RBM20 mutant dilated cardiomyopathy (DCM). In iPSC-CMs harboring a known causal RBM20 variant, the splicing of RBM20 target genes, calcium handling, and contractility are impaired consistent with the disease manifestation in patients. A variant (Pro633Leu) identified by exome sequencing of patient genomes displays the same disease phenotypes, thus establishing this variant as disease causing. We find that all-trans retinoic acid upregulates RBM20 expression and reverts the splicing, calcium handling, and contractility defects in iPSC-CMs with different causal RBM20 mutations. These results suggest that pharmacological upregulation of RBM20 expression is a promising therapeutic strategy for DCM patients with a heterozygous mutation in RBM20.

Keywords: DCM; RBM20; alternative splicing; cardiomyocytes; disease modeling; genome editing; iPSC; precision medicine.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Humans
Induced Pluripotent Stem Cells / metabolism*
RNA Splicing / genetics*
RNA-Binding Proteins / metabolism*
Up-Regulation

Substances

RNA-Binding Proteins
ribonucleic acid binding motif protein 20, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding