TY - JOUR
T1 - Genetic Dissection of a Super Enhancer Controlling the Nppa-Nppb Cluster in the Heart
AU - Man, Joyce Ck
AU - van Duijvenboden, Karel
AU - Krijger, Peter Hl
AU - Hooijkaas, Ingeborg B
AU - van der Made, Ingeborg
AU - de Gier-de Vries, Corrie
AU - Wakker, Vincent
AU - Creemers, Esther E
AU - de Laat, Wouter
AU - Boukens, Bastiaan J
AU - Christoffels, Vincent M
N1 - Funding Information:
This study was supported by CVON HUSTCARE (to V.M. Christoffels), Fonda-tion Leducq 14CVD01 (to V.M. Christoffels), Dutch Heart Foundation CONCOR-genes (to V.M. Christoffels) and the Dutch Heart Foundation 2016T047 (to B.J. Boukens).
Publisher Copyright:
© 2021 Lippincott Williams and Wilkins. All rights reserved.
PY - 2021/1/8
Y1 - 2021/1/8
N2 - RATIONALE: ANP (atrial natriuretic peptide) and BNP (B-type natriuretic peptide), encoded by the clustered genes
Nppa and
Nppb, are important prognostic, diagnostic, and therapeutic proteins in cardiac disease. The spatiotemporal expression pattern and stress-induction of the
Nppa and
Nppb are tightly regulated, possibly involving their coregulation by an evolutionary conserved enhancer cluster.
OBJECTIVE: To explore the physiological functions of the enhancer cluster and elucidate the genomic mechanism underlying
Nppa-Nppb coregulation in vivo.
METHODS AND RESULTS: By analyzing epigenetic data we uncovered an enhancer cluster with super enhancer characteristics upstream of
Nppb. Using CRISPR/Cas9 genome editing, the enhancer cluster or parts thereof,
Nppb and flanking regions or the entire genomic block spanning
Nppa-Nppb, respectively, were deleted from the mouse genome. The impact on gene regulation and phenotype of the respective mouse lines was investigated by transcriptomic, epigenomic, and phenotypic analyses. The enhancer cluster was essential for prenatal and postnatal ventricular expression of
Nppa and
Nppb but not of any other gene. Enhancer cluster-deficient mice showed enlarged hearts before and after birth, similar to
Nppa-Nppb compound knockout mice we generated. Analysis of the other deletion alleles indicated the enhancer cluster engages the promoters of
Nppa and
Nppb in a competitive rather than a cooperative mode, resulting in increased
Nppa expression when
Nppb and flanking sequences were deleted. The enhancer cluster maintained its active epigenetic state and selectivity when its target genes are absent. In enhancer cluster-deficient animals,
Nppa was induced but remained low in the postmyocardial infarction border zone and in the hypertrophic ventricle, involving regulatory sequences proximal to
Nppa.
CONCLUSIONS: Coordinated ventricular expression of
Nppa and
Nppb is controlled in a competitive manner by a shared super enhancer, which is also required to augment stress-induced expression and to prevent premature hypertrophy.
AB - RATIONALE: ANP (atrial natriuretic peptide) and BNP (B-type natriuretic peptide), encoded by the clustered genes
Nppa and
Nppb, are important prognostic, diagnostic, and therapeutic proteins in cardiac disease. The spatiotemporal expression pattern and stress-induction of the
Nppa and
Nppb are tightly regulated, possibly involving their coregulation by an evolutionary conserved enhancer cluster.
OBJECTIVE: To explore the physiological functions of the enhancer cluster and elucidate the genomic mechanism underlying
Nppa-Nppb coregulation in vivo.
METHODS AND RESULTS: By analyzing epigenetic data we uncovered an enhancer cluster with super enhancer characteristics upstream of
Nppb. Using CRISPR/Cas9 genome editing, the enhancer cluster or parts thereof,
Nppb and flanking regions or the entire genomic block spanning
Nppa-Nppb, respectively, were deleted from the mouse genome. The impact on gene regulation and phenotype of the respective mouse lines was investigated by transcriptomic, epigenomic, and phenotypic analyses. The enhancer cluster was essential for prenatal and postnatal ventricular expression of
Nppa and
Nppb but not of any other gene. Enhancer cluster-deficient mice showed enlarged hearts before and after birth, similar to
Nppa-Nppb compound knockout mice we generated. Analysis of the other deletion alleles indicated the enhancer cluster engages the promoters of
Nppa and
Nppb in a competitive rather than a cooperative mode, resulting in increased
Nppa expression when
Nppb and flanking sequences were deleted. The enhancer cluster maintained its active epigenetic state and selectivity when its target genes are absent. In enhancer cluster-deficient animals,
Nppa was induced but remained low in the postmyocardial infarction border zone and in the hypertrophic ventricle, involving regulatory sequences proximal to
Nppa.
CONCLUSIONS: Coordinated ventricular expression of
Nppa and
Nppb is controlled in a competitive manner by a shared super enhancer, which is also required to augment stress-induced expression and to prevent premature hypertrophy.
KW - epigenomics
KW - gene expression regulation
KW - myocardium
KW - natriuretic peptides
KW - regulatory elements, transcriptional
UR - http://www.scopus.com/inward/record.url?scp=85100279827&partnerID=8YFLogxK
U2 - 10.1161/CIRCRESAHA.120.317045
DO - 10.1161/CIRCRESAHA.120.317045
M3 - Article
C2 - 33107387
SN - 0009-7330
VL - 128
SP - 115
EP - 129
JO - Circulation research
JF - Circulation research
IS - 1
ER -