PUBLICATION

Canonical Wnt5b Signaling Directs Outlying Nkx2.5+ Mesoderm into Pacemaker Cardiomyocytes

Authors

Ren, J., Han, P., Ma, X., Farah, E.N., Bloomekatz, J., Zeng, X.I., Zhang, R., Swim, M.M., Witty, A.D., Knight, H.G., Deshpande, R., Xu, W., Yelon, D., Chen, S., Chi, N.C.

ID

ZDB-PUB-190814-2

Date

2019

Source

Developmental Cell 50(6): 729-743.e5 (Journal)

Registered Authors

Bloomekatz, Joshua, Chi, Neil C., Han, Peidong, Yelon, Deborah, Zeng, Sean

Keywords

3D bioprinting, Wnt5b, canonical Wnt signaling, differentiation, human pluripotent stem cells, pacemaker cardiomyocytes, zebrafish

MeSH Terms

Myocytes, Cardiac/cytology
Myocytes, Cardiac/metabolism*
Wnt Proteins/metabolism*
Loss of Function Mutation/genetics
Humans
Gene Expression Regulation, Developmental
Bioprinting
Animals
Stem Cells/metabolism
Zebrafish
Cell Differentiation
Pluripotent Stem Cells/cytology
Pluripotent Stem Cells/metabolism
Base Sequence
Homeobox Protein Nkx-2.5/metabolism*
Mesoderm/metabolism*
Models, Cardiovascular
Signal Transduction*

(all 18)

PubMed

31402282 Full text @ Dev. Cell

Abstract

Pacemaker cardiomyocytes that create the sinoatrial node are essential for the initiation and maintenance of proper heart rhythm. However, illuminating developmental cues that direct their differentiation has remained particularly challenging due to the unclear cellular origins of these specialized cardiomyocytes. By discovering the origins of pacemaker cardiomyocytes, we reveal an evolutionarily conserved Wnt signaling mechanism that coordinates gene regulatory changes directing mesoderm cell fate decisions, which lead to the differentiation of pacemaker cardiomyocytes. We show that in zebrafish, pacemaker cardiomyocytes derive from a subset of Nkx2.5+ mesoderm that responds to canonical Wnt5b signaling to initiate the cardiac pacemaker program, including activation of pacemaker cell differentiation transcription factors Isl1 and Tbx18 and silencing of Nkx2.5. Moreover, applying these developmental findings to human pluripotent stem cells (hPSCs) notably results in the creation of hPSC-pacemaker cardiomyocytes, which successfully pace three-dimensional bioprinted hPSC-cardiomyocytes, thus providing potential strategies for biological cardiac pacemaker therapy.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
Cerulean	co19Tg; fb9Tg	control	Prim-5	cardiac muscle cell	IHC	Fig. 1
EGFP	ia5Tg; twu277Tg	control	20-25 somites	heart primordium cardiac muscle cell	IHC	Fig. 2
EGFP	ia5Tg; twu277Tg	control	14-19 somites	heart primordium cardiac muscle cell	IFL	Fig. 2
EGFP	ia5Tg; twu277Tg	control	20-25 somites	heart primordium cardiac muscle cell	IFL	Fig. 2
Kaede	co19Tg; fb9Tg	control	Prim-5	cardiac conduction system cardiac muscle cell	IHC	Fig. 1
Kaede	co19Tg; fb9Tg	control	Prim-5	cardiac muscle cell	IHC	Fig. 1
Kaede	co19Tg; fb9Tg	control	Prim-5	presumptive atrium heart tube	IHC	Fig. 1
Kaede	co19Tg; fb9Tg	control	Prim-5	presumptive cardiac ventricle heart tube	IHC	Fig. 1
mCherry	apc^sd58/sd58; ia5Tg; twu277Tg	control	Day 4	heart	IFL	Fig. 4
mCherry	fb7Tg; ia5Tg	control	20-25 somites	cardiac conduction system cardiac muscle cell	IHC	Fig. 2

1 - 10 of 28

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
apc^sd58/sd58	control	Long-pec	caudal fin curved dorsal, abnormal	Fig. 4
apc^sd58/sd58	control	Long-pec	pericardial region edematous, abnormal	Fig. 4
apc^sd58/sd58; ia5Tg; twu277Tg	control	Day 4	heart canonical Wnt signaling pathway increased occurrence, abnormal	Fig. 4
apc^sd58/sd58; ia5Tg; twu277Tg	control	Day 4	heart mCherry expression increased amount, abnormal	Fig. 4
apc^sd58/sd58; ia5Tg; twu277Tg	control	Day 4	heart mCherry expression increased distribution, abnormal	Fig. 4
apc^sd58/sd58; sd12Tg	control	Long-pec	heart cardiac muscle cell amount, normal	Fig. 4
apc^sd58/sd58; twu277Tg	control	Long-pec	cardiac conduction system cardiac muscle cell increased amount, abnormal	Fig. 4
co19Tg; w26Tg	heat shock	Prim-5	cardiac conduction system cardiac muscle cell decreased amount, abnormal	Fig. 3
co19Tg; w33Tg	heat shock	Long-pec	cardiac conduction system cardiac muscle cell increased amount, abnormal	Fig. 5
ia5Tg; w33Tg	heat shock	20-25 somites	heart primordium mCherry expression increased amount, abnormal	Fig. 5

1 - 10 of 17

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
co19Tg	Tg(sox10:GAL4-VP16,myl7:Cerulean)	Transgenic Insertion
fb7Tg	TgBAC(-36nkx2.5:ZsYellow)	Transgenic Insertion
fb9Tg	TgBAC(nkx2.5:Kaede)	Transgenic Insertion
fcu1Tg	Tg(hsp70l:nkx2.5-EGFP)	Transgenic Insertion
ia5Tg	Tg(7xTCF-Xla.Sia:NLS-mCherry)	Transgenic Insertion
s878Tg	Tg(myl7:GCaMP)	Transgenic Insertion
s958Tg	Tg(myh6:EGFP)	Transgenic Insertion
sd12Tg	Tg(myl7:h2az2a-mCherry)	Transgenic Insertion
sd58		Point Mutation	apc
ta98		Point Mutation	wnt5b

1 - 10 of 14

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Human Disease / Model

Sequence Targeting Reagents

Fish

Fish
apc^sd58/sd58
apc^sd58/sd58; ia5Tg; twu277Tg
apc^sd58/sd58; sd12Tg
apc^sd58/sd58; twu277Tg
co19Tg; fb9Tg
co19Tg; w26Tg
co19Tg; w33Tg
fb7Tg
fb7Tg; ia5Tg
ia5Tg; twu277Tg

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Cerulean	EFG	Cerulean
EGFP	EFG	EGFP
GAL4	EFG	GAL4
GCaMP	EFG	GCaMP
GFP	EFG	GFP
Kaede	EFG	Kaede
mCherry	EFG	mCherry
ZsYellow	EFG	ZsYellow

Mapping

Ren et al., 2019 ZDB-PUB-190814-2 PMID:31402282

Canonical Wnt5b Signaling Directs Outlying Nkx2.5+ Mesoderm into Pacemaker Cardiomyocytes

Ren et al., 2019

ZDB-PUB-190814-2
PMID:31402282