2. Academic Validation
  3. Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein granules within processing bodies

Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein granules within processing bodies

  • Nat Commun. 2021 Nov 3;12(1):6324. doi: 10.1038/s41467-021-26623-y.
Aidan M Fenix # 1 2 3,  Yuichiro Miyaoka # 4 5,  Alessandro Bertero 1 2 3,  Steven M Blue 6,  Matthew J Spindler 5,  Kenneth K B Tan 5,  Juan A Perez-Bermejo 5,  Amanda H Chan 5,  Steven J Mayerl 5,  Trieu D Nguyen 5,  Caitlin R Russell 5,  Paweena P Lizarraga 5,  Annie Truong 5,  Po-Lin So 5,  Aishwarya Kulkarni 7 8,  Kashish Chetal 8,  Shashank Sathe 6,  Nathan J Sniadecki 1 2 3 9 10,  Gene W Yeo 6,  Charles E Murry 11 12 13 14 15 16,  Bruce R Conklin 17 18,  Nathan Salomonis 19 20
Affiliations

Affiliations

  • 1 Department of Laboratory Medicine and Pathology, University of Washington, 1959 NE Pacific Street, Seattle, WA, 98195, USA.
  • 2 Center for Cardiovascular Biology, University of Washington, 850 Republican Street, Brotman Building, Seattle, WA, 98109, USA.
  • 3 Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Seattle, WA, 98109, USA.
  • 4 Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, 156-8506, Japan.
  • 5 Gladstone Institutes, 1650 Owens St, San Francisco, CA, 94158, USA.
  • 6 Department of Cellular and Molecular Medicine, Stem Cell Program, and Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, 92093, USA.
  • 7 Department of Electrical Engineering and Computer Science, University of Cincinnati, Cincinnati, OH, 45221, USA.
  • 8 Division of Biomedical Informatics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
  • 9 Department of Mechanical Engineering, University of Washington, 3720 15th Avenue NE, Seattle, WA, 98105, USA.
  • 10 Department of Bioengineering, University of Washington, 3720 15th Avenue NE, Seattle, WA, 98105, USA.
  • 11 Department of Laboratory Medicine and Pathology, University of Washington, 1959 NE Pacific Street, Seattle, WA, 98195, USA. murry@uw.edu.
  • 12 Center for Cardiovascular Biology, University of Washington, 850 Republican Street, Brotman Building, Seattle, WA, 98109, USA. murry@uw.edu.
  • 13 Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Seattle, WA, 98109, USA. murry@uw.edu.
  • 14 Department of Bioengineering, University of Washington, 3720 15th Avenue NE, Seattle, WA, 98105, USA. murry@uw.edu.
  • 15 Department of Medicine/Cardiology, University of Washington, 1959 NE Pacific Street, Seattle, WA, 98195, USA. murry@uw.edu.
  • 16 Sana Biotechnology, 188 E Blaine Street, Seattle, WA, 98102, USA. murry@uw.edu.
  • 17 Gladstone Institutes, 1650 Owens St, San Francisco, CA, 94158, USA. bconklin@gladstone.ucsf.edu.
  • 18 Department of Medicine, Cellular and Molecular Pharmacology, and Ophthalmology, University of California San Francisco, San Francisco, CA, 94158, USA. bconklin@gladstone.ucsf.edu.
  • 19 Division of Biomedical Informatics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA. nathan.salomonis@cchmc.org.
  • 20 Department of Pediatrics, University of Cincinnati, Cincinnati, OH, 45229, USA. nathan.salomonis@cchmc.org.
  • # Contributed equally.
PMID: 34732726 DOI: 10.1038/s41467-021-26623-y
Abstract

Mutations in the cardiac splicing factor RBM20 lead to malignant dilated cardiomyopathy (DCM). To understand the mechanism of RBM20-associated DCM, we engineered isogenic iPSCs with DCM-associated missense mutations in RBM20 as well as RBM20 knockout (KO) iPSCs. iPSC-derived engineered heart tissues made from these cell lines recapitulate contractile dysfunction of RBM20-associated DCM and reveal greater dysfunction with missense mutations than KO. Analysis of RBM20 RNA binding by eCLIP reveals a gain-of-function preference of mutant RBM20 for 3' UTR sequences that are shared with amyotrophic lateral sclerosis (ALS) and processing-body associated RNA binding proteins (FUS, DDX6). Deep RNA Sequencing reveals that the RBM20 R636S mutant has unique gene, splicing, polyadenylation and circular RNA defects that differ from RBM20 KO. Super-resolution microscopy verifies that mutant RBM20 maintains very limited nuclear localization potential; rather, the mutant protein associates with cytoplasmic processing bodies (DDX6) under basal conditions, and with stress granules (G3BP1) following acute stress. Taken together, our results highlight a pathogenic mechanism in cardiac disease through splicing-dependent and -independent pathways.

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