PUBLICATION

HHEX is a transcriptional regulator of the VEGFC/FLT4/PROX1 signaling axis during vascular development

Authors

Gauvrit, S., Villasenor, A., Strilic, B., Kitchen, P., Collins, M.M., Marín-Juez, R., Guenther, S., Maischein, H.M., Fukuda, N., Canham, M.A., Brickman, J.M., Bogue, C.W., Jayaraman, P.S., Stainier, D.Y.R.

ID

ZDB-PUB-180715-2

Date

2018

Source

Nature communications 9: 2704 (Journal)

Registered Authors

Maischein, Hans-Martin, Marín-Juez, Rubén, Stainier, Didier

Keywords

none

Datasets

GEO:GSE111963

MeSH Terms

Neovascularization, Physiologic/genetics
Blood Vessels/cytology
Blood Vessels/growth & development
Blood Vessels/metabolism
Animals, Genetically Modified
Zebrafish Proteins/deficiency
Zebrafish Proteins/genetics*
Zebrafish Proteins/metabolism
Embryo, Mammalian
Vascular Endothelial Growth Factor Receptor-3/genetics*
Vascular Endothelial Growth Factor Receptor-3/metabolism
Lymphangiogenesis/genetics*
Lymphatic Vessels/cytology
Lymphatic Vessels/metabolism
Transcription, Genetic
Animals
Tumor Suppressor Proteins/genetics*
Tumor Suppressor Proteins/metabolism
Endothelial Cells/cytology
Endothelial Cells/metabolism
Cell Line
Signal Transduction
Gene Expression Regulation, Developmental*
Mice
Zebrafish
Vascular Endothelial Growth Factor C/genetics*
Vascular Endothelial Growth Factor C/metabolism
Base Sequence
Homeodomain Proteins/genetics*
Homeodomain Proteins/metabolism
Embryo, Nonmammalian
Humans
Repressor Proteins/deficiency
Repressor Proteins/genetics*

(all 34)

PubMed

30006544 Full text @ Nat. Commun.

Abstract

Formation of the lymphatic system requires the coordinated expression of several key regulators: vascular endothelial growth factor C (VEGFC), its receptor FLT4, and a key transcriptional effector, PROX1. Yet, how expression of these signaling components is regulated remains poorly understood. Here, using a combination of genetic and molecular approaches, we identify the transcription factor hematopoietically expressed homeobox (HHEX) as an upstream regulator of VEGFC, FLT4, and PROX1 during angiogenic sprouting and lymphatic formation in vertebrates. By analyzing zebrafish mutants, we found that hhex is necessary for sprouting angiogenesis from the posterior cardinal vein, a process required for lymphangiogenesis. Furthermore, studies of mammalian HHEX using tissue-specific genetic deletions in mouse and knockdowns in cultured human endothelial cells reveal its highly conserved function during vascular and lymphatic development. Our findings that HHEX is essential for the regulation of the VEGFC/FLT4/PROX1 axis provide insights into the molecular regulation of lymphangiogenesis.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
flt4	bns136Tg	standard conditions	Long-pec	whole organism	RTPCR	Fig. S7
flt4	hhex^s722/s722	standard conditions	Prim-5	posterior cardinal vein	ISH	Fig. 2
flt4	hhex^s722/s722	standard conditions	Prim-15	posterior cardinal vein	ISH	Fig. 2
flt4	hhex^s722/s722	standard conditions	Long-pec	whole organism	RTPCR	Fig. S7
hhex	bns136Tg	standard conditions	Long-pec	whole organism	RTPCR	Fig. S7
hhex	hhex^s722/s722	standard conditions	Prim-15	endothelial cell	ISH	Fig. S4
hhex	hhex^s722/s722	standard conditions	Long-pec	whole organism	RTPCR	Fig. S7
hhex	WT	standard conditions	Prim-25	intersegmental vessel	ISH	Fig. S3
hhex	WT	standard conditions	Long-pec	intersegmental vessel	ISH	Fig. S3
hhex	WT	standard conditions	Prim-25	posterior cardinal vein endothelial cell	ISH	Fig. S3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
bns136Tg	standard conditions	Long-pec	whole organism flt4 expression increased amount, abnormal	Fig. S7
bns136Tg	standard conditions	Long-pec	whole organism hhex expression increased amount, abnormal	Fig. S7
bns136Tg	standard conditions	Long-pec	whole organism prox1a expression decreased amount, abnormal	Fig. S7
bns136Tg	standard conditions	Long-pec	whole organism vegfc expression decreased amount, abnormal	Fig. S7
bns136Tg	standard conditions	Protruding-mouth	trunk vasculature morphology, normal	Fig. S6
bns136Tg	standard conditions	Protruding-mouth	vasculature development process quality, normal	Fig. S6
bns136Tg	standard conditions	Day 5	trunk vasculature morphology, normal	Fig. S6
bns136Tg	standard conditions	Day 5	vasculature development process quality, normal	Fig. S6
hhex^s721/s721; ci1Tg	standard conditions	Long-pec	intersegmental vein morphology, normal	Fig. S1
hhex^s721/s721; ci1Tg	standard conditions	Long-pec	lymph vessel endothelium morphology, normal	Fig. S1

1 - 10 of 56

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
b16		Deficiency	hhex ndr2
bns136Tg	Tg(fli1:Tomato-2A-hhex)	Transgenic Insertion
ci1Tg	TgBAC(etsrp:EGFP)	Transgenic Insertion
is4Tg	Tg(kdrl:NLS-mCherry)	Transgenic Insertion
nim5Tg	TgBAC(prox1a:KALTA4,4xUAS-E1B:TagRFP)	Transgenic Insertion
nz101Tg	Tg(-5.2lyve1b:DsRed)	Transgenic Insertion
s721		Insertion	hhex
s722		Small Deletion	hhex
s843Tg	Tg(kdrl:EGFP)	Transgenic Insertion
y7Tg	Tg(fli1:nEGFP)	Transgenic Insertion

1 - 10 of 10

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
hhex	TALEN1-hhex	TALEN

Fish

Fish
bns136Tg
hhex^s721/s721; ci1Tg
hhex^s721/s721; nz101Tg
hhex^s721/s721; nz101Tg; s843Tg
hhex^s721/s721; y7Tg
hhex^s722/s722
hhex^s722/s722; bns136Tg
hhex^s722/s722; ci1Tg
hhex^s722/s722; nim5Tg
hhex^s722/s722; nim5Tg; s843Tg

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab7-prox1	polyclonal	prox1a		Rabbit

Orthology

Gene	Orthology
flt4

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed	EFG	DsRed
EGFP	EFG	EGFP
KALTA4	EFG	KALTA4
mCherry	EFG	mCherry
TagRFP	EFG	TagRFP
Tomato	EFG	Tomato

Mapping

Gauvrit et al., 2018 ZDB-PUB-180715-2 PMID:30006544

HHEX is a transcriptional regulator of the VEGFC/FLT4/PROX1 signaling axis during vascular development

Gauvrit et al., 2018

ZDB-PUB-180715-2
PMID:30006544