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Hierarchical and stage-specific regulation of murine cardiomyocyte maturation by serum response factor

Guo, Yuxuan and Jardin, Blake D. and Zhou, Pingzhu and Sethi, Isha and Akerberg, Brynn N. and Toepfer, Christopher N. and Ai, Yulan and Li, Yifei and Ma, Qing and Guatimosim, Silvia and Hu, Yongwu and Varuzhanyan, Grigor and VanDusen, Nathan J. and Zhang, Donghui and Chan, David C. and Yuan, Guo-Cheng and Seidman, Christine E. and Seidman, Jonathan G. and Pu, William T. (2018) Hierarchical and stage-specific regulation of murine cardiomyocyte maturation by serum response factor. Nature Communications, 9 . Art. No. 3837. ISSN 2041-1723. PMCID PMC6155060. http://resolver.caltech.edu/CaltechAUTHORS:20181001-094858218

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Abstract

After birth, cardiomyocytes (CM) acquire numerous adaptations in order to efficiently pump blood throughout an animal’s lifespan. How this maturation process is regulated and coordinated is poorly understood. Here, we perform a CRISPR/Cas9 screen in mice and identify serum response factor (SRF) as a key regulator of CM maturation. Mosaic SRF depletion in neonatal CMs disrupts many aspects of their maturation, including sarcomere expansion, mitochondrial biogenesis, transverse-tubule formation, and cellular hypertrophy. Maintenance of maturity in adult CMs is less dependent on SRF. This stage-specific activity is associated with developmentally regulated SRF chromatin occupancy and transcriptional regulation. SRF directly activates genes that regulate sarcomere assembly and mitochondrial dynamics. Perturbation of sarcomere assembly but not mitochondrial dynamics recapitulates SRF knockout phenotypes. SRF overexpression also perturbs CM maturation. Together, these data indicate that carefully balanced SRF activity is essential to promote CM maturation through a hierarchy of cellular processes orchestrated by sarcomere assembly.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/s41467-018-06347-2DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6155060/PubMed CentralArticle
ORCID:
AuthorORCID
Guo, Yuxuan0000-0002-6009-4403
Jardin, Blake D.0000-0002-1376-5235
Toepfer, Christopher N.0000-0003-4671-2030
Li, Yifei0000-0002-3096-4287
Varuzhanyan, Grigor0000-0001-6165-0857
Yuan, Guo-Cheng0000-0002-2283-4714
Seidman, Christine E.0000-0001-6380-1209
Pu, William T.0000-0002-4551-8079
Additional Information:© 2018 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received: 14 February 2018 Accepted: 30 August 2018. Published online: 21 September 2018. We thank the HMS EM core, HMS biopolymers core, and DFCI flow cytometry core for technical support. We also thank Drs. Joe Miano and Ivan Moskowitz for sharing the Srf-flox and Tbx5-flox mice, respectively. This work was supported by funding from NIH NHLBI (2UM1 HL098166 and U01HL131003), the American Heart Association (AHA) (17IRG33410894), and charitable support from the Boston Children’s Hospital Department of Cardiology. S.G. was a recipient of a Coordenação de Aperfeiçoamento de Pessoal de Nível Superior fellowship and funded by Conselho Nacional de Desenvolvimento Científico e Tecnológico and Fundação de Amparo a Pesquisa do Estado de Minas Gerais. C.N.T. is a recipient of Sir Henry Wellcome Postdoctoral Fellowship 206466/Z/17/Z. Y.G. is a recipient of AHA postdoctoral fellowship 18POST33960037. Author Contributions: W.T.P. provided overall supervision of this project. Y.G. conceived and designed the study. Y.G., B.D.J., and Y.A. bred animals, produced AAV and performed histology, immunofluorescence, qPCR, and Western blot analysis. Y.G. and B.D.J. performed Langendorff perfusion, in situ heart imaging, flow cytometry, electron microscopy, and mitochondria analysis. Y.L. and D.Z. helped with mitochondria analysis. Y.L. and Q.M. performed echocardiogram analysis. G.V. and D.C.C helped with the analysis of mitofusin mice. N.J.V. helped with animal breeding and AAV production. Y.G. and W.T.P. performed and analyzed RNA-seq. Y.G, W.T.P., P.Z., I.S., B.N.A, and G.-C.Y. performed ChIP-seq and analyzed the data. Y.G., C.N.T., C.E.S., and J.G.S. performed and analyzed cell contractility. Y.G., B.D.J., and S.G. performed calcium imaging and analysis. Y.G. and W.T.P. wrote the manuscript. Data availability: The authors declare that all data supporting the findings of this study are available within the article and its Supplementary information files or from the corresponding author upon reasonable request. RNA-seq and ChIP-seq data have been deposited in the Gene Expression Omnibus (GEO) database under the accession codes: GSE109425 (for the Srf KO RNA-seq), GSE109504 (ChIP-seq), and GSE116030 (for the Srf OE RNA-seq). The data are also available on the Cardiovascular Development Consortium server (https://b2b.hci.utah.edu/gnomex) (sign in as guest). The authors declare no competing interests.
Funders:
Funding AgencyGrant Number
NIH2UM1 HL098166
NIHU01HL131003
American Heart Association17IRG33410894
Boston Children’s HospitalUNSPECIFIED
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)UNSPECIFIED
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)UNSPECIFIED
Fundação de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG)UNSPECIFIED
Wellcome Trust206466/Z/17/Z
American Heart Association18POST33960037
PubMed Central ID:PMC6155060
Record Number:CaltechAUTHORS:20181001-094858218
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20181001-094858218
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:90055
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:03 Oct 2018 17:34
Last Modified:04 Oct 2018 15:14

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