@article{75775f40b78f4a9c8ce86525cb6ff638,
title = "Cardiac Reprogramming Factors Synergistically Activate Genome-wide Cardiogenic Stage-Specific Enhancers",
abstract = "The cardiogenic transcription factors (TFs) Mef2c, Gata4, and Tbx5 can directly reprogram fibroblasts to induced cardiac-like myocytes (iCLMs), presenting a potential source of cells for cardiac repair. While activity of these TFs is enhanced by Hand2 and Akt1, their genomic targets and interactions during reprogramming are not well studied. We performed genome-wide analyses of cardiogenic TF binding and enhancer profiling during cardiac reprogramming. We found that these TFs synergistically activate enhancers highlighted by Mef2c binding sites and that Hand2 and Akt1 coordinately recruit other TFs to enhancer elements. Intriguingly, these enhancer landscapes collectively resemble patterns of enhancer activation during embryonic cardiogenesis. We further constructed a cardiac reprogramming gene regulatory network and found repression of EGFR signaling pathway genes. Consistently, chemical inhibition of EGFR signaling augmented reprogramming. Thus, by defining epigenetic landscapes these findings reveal synergistic transcriptional activation across a broad landscape of cardiac enhancers and key signaling pathways that govern iCLM reprogramming. Hashimoto and colleagues show that reprogramming factors act in concert at cardiac regulatory elements to directly reprogram mouse fibroblasts into induced cardiac-like myocytes (iCLMs). Moreover, cardiac reprogramming is achieved by activation of endogenous cardiac enhancers that initiate a cardiogenic gene regulatory network.",
keywords = "Akt1, Gata4, Hand2, Mef2c, Tbx5, cardiomyocytes, direct reprogramming, heart regeneration, induced cardiac-like myocytes",
author = "Hisayuki Hashimoto and Zhaoning Wang and Garry, {Glynnis A.} and Malladi, {Venkat S.} and Botten, {Giovanni A.} and Wenduo Ye and Huanyu Zhou and Marco Osterwalder and Dickel, {Diane E.} and Axel Visel and Ning Liu and Rhonda Bassel-Duby and Olson, {Eric N.}",
note = "Funding Information: We thank J. Cabrera for graphical assistance; J. McAnally for performing microinjection for the in vivo transgenic reporter assay; J. Richardson, J. Shelton, and the UT Southwestern Histology Core for help with histopathology; and J. Xu, X. Liu, and the Sequencing Core Facility at Children's Research Institute and the Genomics and Microarray Core Facility at UT Southwestern for performing the Illumina sequencing. We are grateful to M.S. Kim for advice about the bioinformatics analysis. This work was supported by grants from the NIH (AR-067294, HL-130253, and HL-138426), Fondation Leducq Transatlantic Networks of Excellence in Cardiovascular Research, and the Robert A. Welch Foundation (grant 1-0025 to E.N.O.). H.H. was supported by a Uehara Memorial Foundation Postdoctoral Fellowship and a Kanae Foreign Study Grant. Z.W. was supported by a predoctoral fellowship from the American Heart Association (19PRE34380436). G.A.G. was supported by an NIH T32 Training grant (5T32HL125247-04). V.S.M was supported by Cancer Prevention and Research Institute of Texas (RP150596). Research by M.O. D.E.D. and A.V. was conducted at the E.O. Lawrence Berkeley National Laboratory and performed under Department of Energy Contract DE-AC02-05CH11231, University of California. The work was supported by NHLBI grant R24HL123879 (to A.V.). We thank the ENCODE Consortium and the ENCODE production laboratories for generating the particular datasets. H.H. Z.W. G.A.G. V.S.M. G.A.B. W.Y. and H.Z. designed and performed experiments and contributed to data analysis, discussion, and writing; M.O. D.E.D. and A.V. contributed to experimental work and discussion; N.L. and R.B.-D. contributed to discussion and writing; and E.N.O. supervised the study and contributed to discussion and writing. E.N.O. is a cofounder and member of the Scientific Advisory Board of Tenaya Therapeutics and holds equity in the company. The other authors declare no competing interests. Funding Information: We thank J. Cabrera for graphical assistance; J. McAnally for performing microinjection for the in vivo transgenic reporter assay; J. Richardson, J. Shelton, and the UT Southwestern Histology Core for help with histopathology; and J. Xu, X. Liu, and the Sequencing Core Facility at Children{\textquoteright}s Research Institute and the Genomics and Microarray Core Facility at UT Southwestern for performing the Illumina sequencing. We are grateful to M.S. Kim for advice about the bioinformatics analysis. This work was supported by grants from the NIH ( AR-067294 , HL-130253 , and HL-138426 ), Fondation Leducq Transatlantic Networks of Excellence in Cardiovascular Research , and the Robert A. Welch Foundation (grant 1-0025 to E.N.O.). H.H. was supported by a Uehara Memorial Foundation Postdoctoral Fellowship and a Kanae Foreign Study Grant . Z.W. was supported by a predoctoral fellowship from the American Heart Association ( 19PRE34380436 ). G.A.G. was supported by an NIH T32 Training grant ( 5T32HL125247-04 ). V.S.M was supported by Cancer Prevention and Research Institute of Texas ( RP150596 ). Research by M.O., D.E.D., and A.V. was conducted at the E.O. Lawrence Berkeley National Laboratory and performed under Department of Energy Contract DE-AC02-05CH11231 , University of California. The work was supported by NHLBI grant R24HL123879 (to A.V.). We thank the ENCODE Consortium and the ENCODE production laboratories for generating the particular datasets. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2019",
month = jul,
day = "3",
doi = "10.1016/j.stem.2019.03.022",
language = "English",
volume = "25",
pages = "69--86.e5",
journal = "Cell Stem Cell",
issn = "1934-5909",
publisher = "Cell Press",
number = "1",
}