Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

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

View ORCID ProfileAidan M. Fenix, View ORCID ProfileYuichiro Miyaoka, View ORCID ProfileAlessandro Bertero, View ORCID ProfileSteven Blue, View ORCID ProfileMatthew J. Spindler, Kenneth K. B. Tan, View ORCID ProfileJuan Perez-Bermejo, Amanda H. Chan, Steven J. Mayer, Trieu Nguyen, Caitlin R. Russell, Paweena Lizarraga, Annie Truong, Po-Lin So, Aishwarya Kulkarni, Kashish Chetal, View ORCID ProfileShashank Sathe, View ORCID ProfileNathan J. Sniadecki, View ORCID ProfileGene W. Yeo, View ORCID ProfileCharles E. Murry, Bruce R. Conklin, View ORCID ProfileNathan Salomonis
doi: https://doi.org/10.1101/2021.06.02.446820
Aidan M. Fenix
1Department of Laboratory Medicine and Pathology, University of Washington, 1959 NE Pacific Street, Seattle, WA 98195, USA
2Center for Cardiovascular Biology, University of Washington, 850 Republican Street, Brotman Building, Seattle, WA 98109, USA
3Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Seattle, WA 98109, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Aidan M. Fenix
Yuichiro Miyaoka
4Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Yuichiro Miyaoka
Alessandro Bertero
1Department of Laboratory Medicine and Pathology, University of Washington, 1959 NE Pacific Street, Seattle, WA 98195, USA
2Center for Cardiovascular Biology, University of Washington, 850 Republican Street, Brotman Building, Seattle, WA 98109, USA
3Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Seattle, WA 98109, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Alessandro Bertero
Steven Blue
8Department of Cellular and Molecular Medicine, Stem Cell Program, and Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Steven Blue
Matthew J. Spindler
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Matthew J. Spindler
Kenneth K. B. Tan
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Juan Perez-Bermejo
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Juan Perez-Bermejo
Amanda H. Chan
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Steven J. Mayer
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Trieu Nguyen
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Caitlin R. Russell
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Paweena Lizarraga
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Annie Truong
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Po-Lin So
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Aishwarya Kulkarni
6Department of Electrical Engineering and Computer Science, University of Cincinnati, Cincinnati, Ohio, 45221, USA
7Division of Biomedical Informatics, Cincinnati Children’s Hospital Medical Center, OH 45229, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kashish Chetal
7Division of Biomedical Informatics, Cincinnati Children’s Hospital Medical Center, OH 45229, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Shashank Sathe
8Department of Cellular and Molecular Medicine, Stem Cell Program, and Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Shashank Sathe
Nathan J. Sniadecki
1Department of Laboratory Medicine and Pathology, University of Washington, 1959 NE Pacific Street, Seattle, WA 98195, USA
2Center for Cardiovascular Biology, University of Washington, 850 Republican Street, Brotman Building, Seattle, WA 98109, USA
3Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Seattle, WA 98109, USA
9Department of Mechanical Engineering, University of Washington, 3720 15th Avenue NE, Seattle, WA 98105, USA
10Department of Bioengineering, University of Washington, 3720 15th Avenue NE, Seattle, WA 98105, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Nathan J. Sniadecki
Gene W. Yeo
8Department of Cellular and Molecular Medicine, Stem Cell Program, and Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Gene W. Yeo
Charles E. Murry
1Department of Laboratory Medicine and Pathology, University of Washington, 1959 NE Pacific Street, Seattle, WA 98195, USA
2Center for Cardiovascular Biology, University of Washington, 850 Republican Street, Brotman Building, Seattle, WA 98109, USA
3Institute for Stem Cell and Regenerative Medicine, University of Washington, 850 Republican Street, Seattle, WA 98109, USA
10Department of Bioengineering, University of Washington, 3720 15th Avenue NE, Seattle, WA 98105, USA
11Department of Medicine/Cardiology, University of Washington, 1959 NE Pacific Street, Seattle, WA 98195, USA
12Sana Biotechnology, 188 E Blaine Street, Seattle, WA 98102, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Charles E. Murry
  • For correspondence: murry@uw.edu bconklin@gladstone.ucsf.edu nathan.salomonis@cchmc.org
Bruce R. Conklin
5Gladstone Institutes, 1650 Owens St. San Francisco CA 94158, USA
13Departments of Medicine, Cellular and Molecular Pharmacology, and Ophthalmology, University of California San Francisco, CA 94158, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: murry@uw.edu bconklin@gladstone.ucsf.edu nathan.salomonis@cchmc.org
Nathan Salomonis
7Division of Biomedical Informatics, Cincinnati Children’s Hospital Medical Center, OH 45229, USA
14Departments of Pediatrics, University of Cincinnati, OH 45229, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Nathan Salomonis
  • For correspondence: murry@uw.edu bconklin@gladstone.ucsf.edu nathan.salomonis@cchmc.org
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Data/Code
  • Preview PDF
Loading

ABSTRACT

RNA binding motif protein 20 (RBM20) is a key regulator of alternative splicing in the heart, and its mutation leads to malignant dilated cardiomyopathy (DCM). To understand the mechanism of RBM20-associated DCM, we engineered isogenic human induced pluripotent stem cells (iPSCs) with heterozygous or homozygous DCM-associated missense mutations in RBM20 (R636S) as well as RBM20 knockout (KO) iPSCs. iPSC-derived engineered heart tissues made from these cell lines recapitulated contractile dysfunction of RBM20-associated DCM and revealed greater dysfunction with missense mutations than KO. Analysis of RBM20 RNA binding by eCLIP revealed 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 revealed that the RBM20 R636S mutant has unique gene, splicing, polyadenylation and circular RNA defects that differ from RBM20 KO, impacting distinct cardiac signaling pathways. Splicing defects specific to KO or R636S mutations were supported by data from R636S gene-edited pig hearts and eCLIP. Super-resolution microscopy verified that mutant RBM20 maintains 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 novel pathogenic mechanism in cardiac disease through splicing-dependent and -independent pathways that are likely to mediate differential contractile phenotypes and stress-associated heart pathology.

Competing Interest Statement

N.J.S. is a scientific advisor to and has equity in Curi Bio, Inc. C.E.M. is a scientific founder and equity holder in Sana Biotechnology. The other authors declare no competing interests.

Footnotes

  • Disclosures N.J.S. is a scientific advisor to and has equity in Curi Bio, Inc. C.E.M. is a scientific founder and equity holder in Sana Biotechnology. The other authors declare no competing interests.

  • https://www.synapse.org/#!Synapse:syn2582579

  • https://github.com/nsalomonis/altanalyze

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
Back to top
PreviousNext
Posted June 03, 2021.
Download PDF

Supplementary Material

Data/Code
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein aggregates within processing bodies
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein aggregates within processing bodies
Aidan M. Fenix, Yuichiro Miyaoka, Alessandro Bertero, Steven Blue, Matthew J. Spindler, Kenneth K. B. Tan, Juan Perez-Bermejo, Amanda H. Chan, Steven J. Mayer, Trieu Nguyen, Caitlin R. Russell, Paweena Lizarraga, Annie Truong, Po-Lin So, Aishwarya Kulkarni, Kashish Chetal, Shashank Sathe, Nathan J. Sniadecki, Gene W. Yeo, Charles E. Murry, Bruce R. Conklin, Nathan Salomonis
bioRxiv 2021.06.02.446820; doi: https://doi.org/10.1101/2021.06.02.446820
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein aggregates within processing bodies
Aidan M. Fenix, Yuichiro Miyaoka, Alessandro Bertero, Steven Blue, Matthew J. Spindler, Kenneth K. B. Tan, Juan Perez-Bermejo, Amanda H. Chan, Steven J. Mayer, Trieu Nguyen, Caitlin R. Russell, Paweena Lizarraga, Annie Truong, Po-Lin So, Aishwarya Kulkarni, Kashish Chetal, Shashank Sathe, Nathan J. Sniadecki, Gene W. Yeo, Charles E. Murry, Bruce R. Conklin, Nathan Salomonis
bioRxiv 2021.06.02.446820; doi: https://doi.org/10.1101/2021.06.02.446820

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Cell Biology
Subject Areas
All Articles
  • Animal Behavior and Cognition (4682)
  • Biochemistry (10357)
  • Bioengineering (7670)
  • Bioinformatics (26330)
  • Biophysics (13523)
  • Cancer Biology (10683)
  • Cell Biology (15438)
  • Clinical Trials (138)
  • Developmental Biology (8497)
  • Ecology (12820)
  • Epidemiology (2067)
  • Evolutionary Biology (16851)
  • Genetics (11399)
  • Genomics (15478)
  • Immunology (10616)
  • Microbiology (25207)
  • Molecular Biology (10220)
  • Neuroscience (54463)
  • Paleontology (401)
  • Pathology (1668)
  • Pharmacology and Toxicology (2897)
  • Physiology (4342)
  • Plant Biology (9243)
  • Scientific Communication and Education (1586)
  • Synthetic Biology (2557)
  • Systems Biology (6780)
  • Zoology (1466)