TY - JOUR
T1 - Hand2 Selectively Reorganizes Chromatin Accessibility to Induce Pacemaker-like Transcriptional Reprogramming
AU - Fernandez-Perez, Antonio
AU - Sathe, Adwait Amod
AU - Bhakta, Minoti
AU - Leggett, Kayla
AU - Xing, Chao
AU - Munshi, Nikhil Vilas
N1 - Funding Information:
We thank the UT Southwestern Medical Center Flow Cytometry, Histology Core, Shared Molecular Biology Confocal Microscopy, and Next Generation Sequencing Facilities for use of resources and expert technical assistance. We gratefully acknowledge Gary Hon, Ning Liu, and Beverly Rothermel for critical review of the manuscript and members of the Olson, Hon, and Munshi labs for technical advice, reagents, and scientific discussions. This work was supported by the NSF (grant 2015165336 to A.F.-P.), NIH (HL136604, HL133642, and HL135217 to N.V.M.), the Burroughs Wellcome Fund (1009838 to N.V.M.), and the March of Dimes Foundation (5-FY13-203 to N.V.M.). Conceptualization, N.V.M.; Experimental Investigation, A.F.-P. M.B. and K.L.; Bioinformatic Analysis, A.A.S. and C.X.; Writing – Original Draft, N.V.M. and A.F.-P.; Writing – Review & Editing, N.V.M. and A.F.-P.; Supervision, N.V.M. and C.X.; and Funding Acquisition, N.V.M. and A.F.-P. The authors declare no competing interests.
Funding Information:
We thank the UT Southwestern Medical Center Flow Cytometry, Histology Core, Shared Molecular Biology Confocal Microscopy, and Next Generation Sequencing Facilities for use of resources and expert technical assistance. We gratefully acknowledge Gary Hon, Ning Liu, and Beverly Rothermel for critical review of the manuscript and members of the Olson, Hon, and Munshi labs for technical advice, reagents, and scientific discussions. This work was supported by the NSF (grant 2015165336 to A.F.-P.), NIH ( HL136604 , HL133642 , and HL135217 to N.V.M.), the Burroughs Wellcome Fund ( 1009838 to N.V.M.), and the March of Dimes Foundation ( 5-FY13-203 to N.V.M.).
Publisher Copyright:
© 2019 The Author(s)
PY - 2019/5/21
Y1 - 2019/5/21
N2 - Gata4, Hand2, Mef2c, and Tbx5 (GHMT) can reprogram transduced fibroblasts into induced pacemaker-like myocytes (iPMs), but the underlying mechanisms remain obscure. Here, we explore the role of Hand2 in iPM formation by using a combination of transcriptome, genome, and biochemical assays. We found many shared transcriptional signatures between iPMs and the endogenous sinoatrial node (SAN), yet key regulatory networks remain missing. We demonstrate that Hand2 augments chromatin accessibility at loci involved in sarcomere organization, electrical coupling, and membrane depolarization. Focusing on an established cardiac Hand2 cistrome, we observe selective reorganization of chromatin accessibility to promote pacemaker-specific gene expression. Moreover, we identify a Hand2 cardiac subtype diversity (CSD) domain through biochemical analysis of the N terminus. By integrating our RNA-seq and ATAC-seq datasets, we highlight desmosome organization as a hallmark feature of iPM formation. Collectively, our results illuminate Hand2-dependent mechanisms that may guide future efforts to rationally improve iPM formation.
AB - Gata4, Hand2, Mef2c, and Tbx5 (GHMT) can reprogram transduced fibroblasts into induced pacemaker-like myocytes (iPMs), but the underlying mechanisms remain obscure. Here, we explore the role of Hand2 in iPM formation by using a combination of transcriptome, genome, and biochemical assays. We found many shared transcriptional signatures between iPMs and the endogenous sinoatrial node (SAN), yet key regulatory networks remain missing. We demonstrate that Hand2 augments chromatin accessibility at loci involved in sarcomere organization, electrical coupling, and membrane depolarization. Focusing on an established cardiac Hand2 cistrome, we observe selective reorganization of chromatin accessibility to promote pacemaker-specific gene expression. Moreover, we identify a Hand2 cardiac subtype diversity (CSD) domain through biochemical analysis of the N terminus. By integrating our RNA-seq and ATAC-seq datasets, we highlight desmosome organization as a hallmark feature of iPM formation. Collectively, our results illuminate Hand2-dependent mechanisms that may guide future efforts to rationally improve iPM formation.
KW - chromatin accessibility
KW - desmosome
KW - gene expression
KW - pacemaker
KW - reprogramming
KW - transcriptome
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UR - http://www.scopus.com/inward/citedby.url?scp=85065120129&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2019.04.077
DO - 10.1016/j.celrep.2019.04.077
M3 - Article
C2 - 31116981
AN - SCOPUS:85065120129
SN - 2211-1247
VL - 27
SP - 2354-2369.e7
JO - Cell Reports
JF - Cell Reports
IS - 8
ER -