PUBLICATION

Positive Feedback Defines the Timing, Magnitude, and Robustness of Angiogenesis

Authors

Page, D.J., Thuret, R., Venkatraman, L., Takahashi, T., Bentley, K., Herbert, S.P.

ID

ZDB-PUB-190613-12

Date

2019

Source

Cell Reports 27: 3139-3151.e5 (Journal)

Registered Authors

Herbert, Shane, Takahashi, Tokiharu, Thuret, Raphael

Keywords

angiogenesis, endothelial cell, lateral inhibition, positive feedback, tetraspanin, tip cell

MeSH Terms

Animals
Humans
Tetraspanins/genetics
Tetraspanins/metabolism
Human Umbilical Vein Endothelial Cells/metabolism
Human Umbilical Vein Endothelial Cells/physiology
Zebrafish
Receptors, Notch/metabolism
Neovascularization, Physiologic*
Feedback, Physiological*
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism
Vascular Endothelial Growth Factor A/metabolism

(all 13)

PubMed

31189101 Full text @ Cell Rep.

Abstract

Angiogenesis is driven by the coordinated collective branching of specialized leading "tip" and trailing "stalk" endothelial cells (ECs). While Notch-regulated negative feedback suppresses excessive tip selection, roles for positive feedback in EC identity decisions remain unexplored. Here, by integrating computational modeling with in vivo experimentation, we reveal that positive feedback critically modulates the magnitude, timing, and robustness of angiogenic responses. In silico modeling predicts that positive-feedback-mediated amplification of VEGF signaling generates an ultrasensitive bistable switch that underpins quick and robust tip-stalk decisions. In agreement, we define a positive-feedback loop exhibiting these properties in vivo, whereby Vegf-induced expression of the atypical tetraspanin, tm4sf18, amplifies Vegf signaling to dictate the speed and robustness of EC selection for angiogenesis. Consequently, tm4sf18 mutant zebrafish select fewer motile ECs and exhibit stunted hypocellular vessels with unstable tip identity that is severely perturbed by even subtle Vegfr attenuation. Hence, positive feedback spatiotemporally shapes the angiogenic switch to ultimately modulate vascular network topology.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
avpr1ab	s843Tg	control	Prim-15	endothelial cell	OTHR	Fig. 3
avpr1ab	s843Tg	chemical treatment: DAPT, chemical treatment: semaxanib	Prim-15	endothelial cell	OTHR	Fig. 3
avpr1ab	s843Tg	chemical treatment: semaxanib	Prim-15	endothelial cell	OTHR	Fig. 3
avpr1ab	s843Tg	chemical treatment: DAPT	Prim-15	endothelial cell	OTHR	Fig. 3
ccl38.1	s843Tg	chemical treatment: DAPT	Prim-15	endothelial cell	OTHR	Fig. 3
ccl38.1	s843Tg	control	Prim-15	endothelial cell	OTHR	Fig. 3
ccl38.1	s843Tg	chemical treatment: semaxanib	Prim-15	endothelial cell	OTHR	Fig. 3
ccl38.1	s843Tg	chemical treatment: DAPT, chemical treatment: semaxanib	Prim-15	endothelial cell	OTHR	Fig. 3
dll4	WT	chemical treatment: semaxanib	Prim-15	whole organism	RTPCR	Fig. 3
EGFP	s843Tg	standard conditions	26+ somites	dorsal aorta	IFL	Fig. 3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment: hydroxyurea, chemical treatment: aphidicolin	26+ somites to Prim-15	endothelial cell angioblast cell migration decreased linear velocity, abnormal	Fig. 6
WT	chemical treatment: hydroxyurea, chemical treatment: aphidicolin	26+ somites to Prim-15	endothelial tip cell angioblast cell migration decreased linear velocity, abnormal	Fig. 6
WT	chemical treatment: hydroxyurea, chemical treatment: aphidicolin	26+ somites to Prim-15	intersegmental vein increased amount, abnormal	Fig. 5
WT	chemical treatment: semaxanib	Prim-15	whole organism dll4 expression decreased amount, abnormal	Fig. 3
WT	chemical treatment: semaxanib	Prim-15	whole organism flt4 expression decreased amount, abnormal	Fig. 3
WT	chemical treatment: semaxanib	Prim-15	whole organism kdrl expression decreased amount, abnormal	Fig. 3
WT	chemical treatment: semaxanib	Prim-15	whole organism tm4sf18 expression decreased amount, abnormal	Fig. 3
WT + MO1-dll4	standard conditions	26+ somites	dorsal aorta tm4sf18 expression increased distribution, abnormal	Fig. 3
WT + MO1-dll4	standard conditions	Prim-15	whole organism kdrl expression increased amount, abnormal	Fig. 3
WT + MO1-dll4	standard conditions	Prim-15	whole organism tm4sf18 expression increased amount, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
s5		Point Mutation	npas4l
s843Tg	Tg(kdrl:EGFP)	Transgenic Insertion
ubs1Tg	Tg(kdrl:NLS-EGFP)	Transgenic Insertion
umc201		Indel	tm4sf18
umc202		Small Deletion	tm4sf18

1 - 5 of 5

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
dll4	MO1-dll4	MRPHLNO
flt1	MO3-flt1	MRPHLNO
tm4sf18	CRISPR1-tm4sf18	CRISPR
tm4sf18	TALEN1-tm4sf18	TALEN

Fish

Fish
npas4l^s5/s5
s843Tg
tm4sf18^umc201/+
tm4sf18^{umc201/umc201}
tm4sf18^{umc201/umc201}; ubs1Tg
ubs1Tg
ubs1Tg + MO1-dll4
ubs1Tg + MO3-flt1
WT
WT + MO1-dll4

Antibodies

Name	Type	Isotypes	Host Organism
Ab9-mapk	monoclonal	IgG	Rabbit
Ab11-mapk	monoclonal	IgG	Rabbit
Ab20-mapk	monoclonal	IgG	Rabbit

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP

Mapping

Page et al., 2019 ZDB-PUB-190613-12 PMID:31189101

Positive Feedback Defines the Timing, Magnitude, and Robustness of Angiogenesis

Page et al., 2019

ZDB-PUB-190613-12
PMID:31189101