NKX2-5 regulates human cardiomyogenesis via a HEY2 dependent transcriptional network

David J Anderson; David I Kaplan; Katrina M Bell; Katerina Koutsis; John M Haynes; Richard J Mills; Dean G Phelan; Elizabeth L Qian; Ana Rita Leitoguinho; Deevina Arasaratnam; Tanya Labonne; Elizabeth S Ng; Richard P Davis; Simona Casini; Robert Passier; James E Hudson; Enzo R Porrello; Mauro W Costa; Arash Rafii; Clare L Curl; Lea M Delbridge; Richard P Harvey; Alicia Oshlack; Michael M Cheung; Christine L Mummery; Stephen Petrou; Andrew G Elefanty; Edouard G Stanley; David A Elliott

doi:10.1038/s41467-018-03714-x

NKX2-5 regulates human cardiomyogenesis via a HEY2 dependent transcriptional network

Nat Commun. 2018 Apr 10;9(1):1373. doi: 10.1038/s41467-018-03714-x.

Authors

David J Anderson¹, David I Kaplan², Katrina M Bell¹, Katerina Koutsis¹, John M Haynes³, Richard J Mills⁴, Dean G Phelan¹, Elizabeth L Qian¹, Ana Rita Leitoguinho¹, Deevina Arasaratnam¹, Tanya Labonne¹, Elizabeth S Ng¹, Richard P Davis⁵, Simona Casini⁵, Robert Passier⁵, James E Hudson⁴, Enzo R Porrello⁴, Mauro W Costa⁶, Arash Rafii^{7

8}, Clare L Curl⁹, Lea M Delbridge⁹, Richard P Harvey^{10

11}, Alicia Oshlack¹, Michael M Cheung^{1

12}, Christine L Mummery⁵, Stephen Petrou², Andrew G Elefanty^{1

12

13}, Edouard G Stanley^{1

12

13}, David A Elliott^{14

15

16}

Affiliations

¹ Murdoch Childrens Research Institute, Royal Children's Hospital, Flemington Road, Parkville, VIC, 3052, Australia.
² The Florey Institute of Neuroscience and Mental Health; Centre for Neuroscience, University of Melbourne, Parkville, VIC, 3052, Australia.
³ Monash Institute of Pharmaceutical Science, Monash University, 381 Royal Parade Parkville, Victoria, 3052, Australia.
⁴ School of Biomedical Sciences, University of Queensland, Brisbane, QLD, 4072, Australia.
⁵ Department of Anatomy and Embryology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, The Netherlands.
⁶ The Jackson Laboratory, Bar Harbor, ME, 04609, USA.
⁷ Stem Cell and Microenvironment Laboratory, Weill Cornell Medical College in Qatar Qatar Foundation, Doha, Qatar.
⁸ Department of Genetic Medicine, Weill Cornell Medical College, New York, NY, USA.
⁹ Department of Physiology, University of Melbourne, Parkville, VIC, 3052, Australia.
¹⁰ Victor Chang Cardiac Research Institute, Darlinghurst, NSW, 2052, Australia.
¹¹ St. Vincent's Clinical School and School of Biotechnology and Biomolecular Sciences, University of New South Wales, Kensington, 2052, Australia.
¹² Department of Pediatrics, The Royal Children's Hospital, University of Melbourne, Parkville, VIC, 3052, Australia.
¹³ Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Clayton, VIC, 3800, Australia.
¹⁴ Murdoch Childrens Research Institute, Royal Children's Hospital, Flemington Road, Parkville, VIC, 3052, Australia. [email protected].
¹⁵ Australian Regenerative Medicine Institute, Monash University, Clayton, VIC, 3800, Australia. [email protected].
¹⁶ School of Biosciences, University of Melbourne, Parkville, VIC, 3052, Australia. [email protected].

Abstract

Congenital heart defects can be caused by mutations in genes that guide cardiac lineage formation. Here, we show deletion of NKX2-5, a critical component of the cardiac gene regulatory network, in human embryonic stem cells (hESCs), results in impaired cardiomyogenesis, failure to activate VCAM1 and to downregulate the progenitor marker PDGFRα. Furthermore, NKX2-5 null cardiomyocytes have abnormal physiology, with asynchronous contractions and altered action potentials. Molecular profiling and genetic rescue experiments demonstrate that the bHLH protein HEY2 is a key mediator of NKX2-5 function during human cardiomyogenesis. These findings identify HEY2 as a novel component of the NKX2-5 cardiac transcriptional network, providing tangible evidence that hESC models can decipher the complex pathways that regulate early stage human heart development. These data provide a human context for the evaluation of pathogenic mutations in congenital heart disease.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Action Potentials / physiology
Basic Helix-Loop-Helix Transcription Factors / genetics*
Basic Helix-Loop-Helix Transcription Factors / metabolism
Cell Differentiation
Cell Line
Gene Deletion
Gene Expression Regulation, Developmental
Gene Regulatory Networks*
Homeobox Protein Nkx-2.5 / deficiency
Homeobox Protein Nkx-2.5 / genetics*
Human Embryonic Stem Cells / cytology
Human Embryonic Stem Cells / metabolism*
Humans
Myocardium / cytology
Myocardium / metabolism
Myocytes, Cardiac / cytology
Myocytes, Cardiac / metabolism*
Organogenesis / genetics*
Patch-Clamp Techniques
Receptor, Platelet-Derived Growth Factor alpha / genetics
Receptor, Platelet-Derived Growth Factor alpha / metabolism
Repressor Proteins / genetics*
Repressor Proteins / metabolism
Transcription, Genetic
Vascular Cell Adhesion Molecule-1 / genetics
Vascular Cell Adhesion Molecule-1 / metabolism

Substances

Basic Helix-Loop-Helix Transcription Factors
HEY2 protein, human
Homeobox Protein Nkx-2.5
NKX2-5 protein, human
Repressor Proteins
Vascular Cell Adhesion Molecule-1
Receptor, Platelet-Derived Growth Factor alpha