@article{a00765f3e5d14434b3e7f204ea870348,
title = "Functional screening in human cardiac organoids reveals a metabolic mechanism for cardiomyocyte cell cycle arrest",
abstract = "The mammalian heart undergoes maturation during postnatal life to meet the increased functional requirements of an adult. However, the key drivers of this process remain poorly defined. We are currently unable to recapitulate postnatal maturation in human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs), limiting their potential as a model system to discover regenerative therapeutics. Here, we provide a summary of our studies, where we developed a 96-well device for functional screening in human pluripotent stem cell-derived cardiac organoids (hCOs). Through interrogation of >10,000 organoids, we systematically optimize parameters, including extracellular matrix (ECM), metabolic substrate, and growth factor conditions, that enhance cardiac tissue viability, function, and maturation. Under optimized maturation conditions, functional and molecular characterization revealed that a switch to fatty acid metabolism was a central driver of cardiac maturation. Under these conditions, hPSC-CMs were refractory to mitogenic stimuli, and we found that key proliferation pathways including β-catenin and Yes-associated protein 1 (YAP1) were repressed. This proliferative barrier imposed by fatty acid metabolism in hCOs could be rescued by simultaneous activation of both β-catenin and YAP1 using genetic approaches or a small molecule activating both pathways. These studies highlight that human organoids coupled with higher-throughput screening platforms have the potential to rapidly expand our knowledge of human biology and potentially unlock therapeutic strategies.",
keywords = "Heart development, Metabolism, Pluripotent stem cells, Regeneration, Tissue engineering",
author = "Mills, {Richard J.} and Titmarsh, {Drew M.} and Xaver Koenig and Parker, {Benjamin L.} and Ryall, {James G.} and Quaife-Ryan, {Gregory A.} and Voges, {Holly K.} and Hodson, {Mark P.} and Charles Ferguson and Lauren Drowley and Plowright, {Alleyn T.} and Needham, {Elise J.} and Wang, {Qing Dong} and Paul Gregorevic and Mei Xin and Thomas, {Walter G.} and Parton, {Robert G.} and Nielsen, {Lars K.} and Launikonis, {Bradley S.} and James, {David E.} and Elliott, {David A.} and Porrello, {Enzo R.} and Hudson, {James E.}",
note = "Funding Information: ACKNOWLEDGMENTS. We thank Shaun Walters for his assistance in setting up the customized Leica DMI8 microscope for our applications and Dr. Sean Lal (Sydney Heart Bank) for providing the human heart biopsy for proteomic analysis. We thank the Developmental Studies Hybridoma Bank for providing the β-catenin (PY489) and titin antibodies. We used the Australian National Fabrication Facility Queensland Node for the fabrication of the Heart-Dyno molds. We also thank QFAB bioinformatics for access to MetaCore as well as the GVL project and the Research Computing Centre for access to the Galaxy-qld server (https://galaxy-qld.genome.edu.au/galaxy). We acknowledge the use of the Australian Microscopy & Microanalysis Research Facility at the Center for Microscopy and Microanalysis at The University of Queensland. R.G.P. was supported by National Health and Medical Research Council of Australia Grants APP1037320, APP1058565, and APP569542 and the Australian Research Council Centre of Excellence in Convergent Bio-Nano Science and Technology. D.A.E. and E.R.P. are supported by the Victorian Government{\textquoteright}s Operational Infrastructure Support Program. E.R.P. and J.E.H. are supported by fellowships and project grants from the National Health and Medical Research Council, the National Heart Foundation, Stem Cells Australia, and The University of Queensland. Publisher Copyright: {\textcopyright} 2017, National Academy of Sciences. All rights reserved.",
year = "2017",
month = oct,
day = "3",
doi = "10.1073/pnas.1707316114",
language = "English (US)",
volume = "114",
pages = "E8372--E8381",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "40",
}