PUBLICATION

Chemical Modulators of Fibrinogen Production and Their Impact on Venous Thrombosis

Authors

Vilar, R., Lukowski, S.W., Garieri, M., Di Sanza, C., Neerman-Arbez, M., Fish, R.J.

ID

ZDB-PUB-201212-26

Date

2020

Source

Thrombosis and haemostasis 121(4): 433-448 (Journal)

Registered Authors

di Sanza, Corinne, Fish, Richard, Lukowski, Sam, Neerman-Arbez, Marguerite

Keywords

none

MeSH Terms

Fibrinolytic Agents/pharmacology*
Hepatocytes/drug effects
Hepatocytes/metabolism
Mutation
Humans
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Anthralin/pharmacology
Zebrafish/genetics
Hep G2 Cells
Fibrinogen/genetics
Fibrinogen/metabolism*
Venous Thrombosis/drug therapy*
Venous Thrombosis/genetics
Venous Thrombosis/metabolism
Animals, Genetically Modified
Blood Coagulation/drug effects*
Tretinoin/pharmacology
Small Molecule Libraries
Proteomics
Morpholinos/pharmacology
Integrin alpha2/genetics
Integrin alpha2/metabolism
Animals
Disease Models, Animal
Oligonucleotides, Antisense/pharmacology

(all 26)

PubMed

33302304 Full text @ Thromb. Haemost.

Abstract

Thrombosis is a leading cause of morbidity and mortality. Fibrinogen, the soluble substrate for fibrin-based clotting, has a central role in haemostasis and thrombosis and its plasma concentration correlates with cardiovascular disease event risk and a prothrombotic state in experimental models. We aimed to identify chemical entities capable of changing fibrinogen production and test their impact on experimental thrombosis. A total of 1,280 bioactive compounds were screened for their ability to alter fibrinogen production by hepatocyte-derived cancer cells and a selected panel was tested in zebrafish larvae. Anthralin and all-trans retinoic acid (RA) were identified as fibrinogen-lowering and fibrinogen-increasing moieties, respectively. In zebrafish larvae, anthralin prolonged laser-induced venous- occlusion times and reduced thrombocyte accumulation at injury sites. RA had opposite effects. Treatment with RA, a nuclear receptor ligand, increased fibrinogen mRNA levels. Using an antisense morpholino oligonucleotide to deplete zebrafish fibrinogen, we correlated a shortening of laser-induced venous thrombosis times with RA treatment and fibrinogen protein levels. Anthralin had little effect on fibrinogen mRNA in zebrafish larvae, despite leading to lower detectable fibrinogen. Therefore, we made a proteomic scan of anthralin-treated cells and larvae. A reduced representation of proteins linked to the canonical secretory pathway was detected, suggesting that anthralin affects protein secretion. In summary, we found that chemical modulation of fibrinogen levels correlates with measured effects on experimental venous thrombosis and could be investigated as a therapeutic avenue for thrombosis prevention.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
fga	TU	chemical treatment by environment: loteprednol	Day 5	whole organism	WB	Fig. 3
fga	TU	chemical treatment by environment: retinoic acid	Day 5	whole organism	WB	Fig. 3
fga	TU	standard conditions	Protruding-mouth	whole organism	WB	Fig. 3
fga	TU	chemical treatment by environment: anthralin	Protruding-mouth	whole organism	WB	Fig. 3
fga	TU	standard conditions	Day 5	whole organism	WB	Fig. 3
fga	TU	chemical treatment by environment: anthralin	Day 5	whole organism	WB	Fig. 3
fga	TU	chemical treatment by environment: loteprednol	Protruding-mouth	whole organism	WB	Fig. 3
fga	TU	chemical treatment by environment: retinoic acid	Protruding-mouth	whole organism	WB	Fig. 3
fga	TU	standard conditions	Day 5	whole organism	RTPCR	Fig. 6
fga	TU	chemical treatment by environment: retinoic acid	Day 5	whole organism	RTPCR	Fig. 6

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
TU	chemical treatment by environment: anthralin	Protruding-mouth	whole organism fga expression decreased amount, abnormal	Fig. 3
TU	chemical treatment by environment: loteprednol	Day 5	whole organism fga expression decreased amount, abnormal	Fig. 3
TU	chemical treatment by environment: retinoic acid	Day 5	whole organism fga expression increased amount, abnormal	Fig. 3
TU	chemical treatment by environment: retinoic acid	Day 5	whole organism fga expression increased amount, abnormal	Fig. 6
TU	chemical treatment by environment: retinoic acid	Day 5	whole organism fgb expression increased amount, abnormal	Fig. 6
TU	chemical treatment by environment: retinoic acid	Day 5	whole organism fgg expression increased amount, abnormal	Fig. 6
la2Tg (TU)	chemical treatment by environment: anthralin	Protruding-mouth	posterior cardinal vein platelet aggregation decreased occurrence, abnormal	Fig. 5
la2Tg (TU)	chemical treatment by environment: anthralin	Day 5	posterior cardinal vein platelet aggregation decreased occurrence, abnormal	Fig. 4
la2Tg (TU)	chemical treatment by environment: retinoic acid	Protruding-mouth	posterior cardinal vein platelet aggregation increased rate, abnormal	Fig. 5
la2Tg (TU)	chemical treatment by environment: retinoic acid	Day 5	posterior cardinal vein platelet aggregation increased occurrence, abnormal	Fig. 4

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
la2Tg	Tg(-6.0itga2b:EGFP)	Transgenic Insertion
ug701		Indel	fga

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
fga	MO1-fga	MRPHLNO

Fish

Fish
fga^ug701/ug701 (TU)
la2Tg (TU)
TU
TU + MO1-fga

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP

Mapping

Vilar et al., 2020 ZDB-PUB-201212-26 PMID:33302304

Chemical Modulators of Fibrinogen Production and Their Impact on Venous Thrombosis

Vilar et al., 2020

ZDB-PUB-201212-26
PMID:33302304