IRUS Total

Zebrafish model for functional screening of flow-responsive genes

File Description SizeFormat 
253637_0_art_file_1989167_p3pfvj_convrt.pdfAccepted version7.05 MBAdobe PDFView/Open
Title: Zebrafish model for functional screening of flow-responsive genes
Authors: Serbanovic-Canic, J
De Luca, A
Warboys, C
Ferreira, PF
Luong, LA
Hsiao, S
Gauci, I
Mahmoud, M
Feng, S
Souilhol, C
Bowden, N
Ashton, JP
Walczak, H
Firmin, D
Krams, R
Mason, JC
Haskard, DO
Sherwin, S
Ridger, V
Chico, TJ
Evans, PC
Item Type: Journal Article
Abstract: OBJECTIVE: Atherosclerosis is initiated at branches and bends of arteries exposed to disturbed blood flow that generates low shear stress. This mechanical environment promotes lesions by inducing endothelial cell (EC) apoptosis and dysfunction via mechanisms that are incompletely understood. Although transcriptome-based studies have identified multiple shear-responsive genes, most of them have an unknown function. To address this, we investigated whether zebrafish embryos can be used for functional screening of mechanosensitive genes that regulate EC apoptosis in mammalian arteries. APPROACH AND RESULTS: First, we demonstrated that flow regulates EC apoptosis in developing zebrafish vasculature. Specifically, suppression of blood flow in zebrafish embryos (by targeting cardiac troponin) enhanced that rate of EC apoptosis (≈10%) compared with controls exposed to flow (≈1%). A panel of candidate regulators of apoptosis were identified by transcriptome profiling of ECs from high and low shear stress regions of the porcine aorta. Genes that displayed the greatest differential expression and possessed 1 to 2 zebrafish orthologues were screened for the regulation of apoptosis in zebrafish vasculature exposed to flow or no-flow conditions using a knockdown approach. A phenotypic change was observed in 4 genes; p53-related protein (PERP) and programmed cell death 2-like protein functioned as positive regulators of apoptosis, whereas angiopoietin-like 4 and cadherin 13 were negative regulators. The regulation of perp, cdh13, angptl4, and pdcd2l by shear stress and the effects of perp and cdh13 on EC apoptosis were confirmed by studies of cultured EC exposed to flow. CONCLUSIONS: We conclude that a zebrafish model of flow manipulation coupled to gene knockdown can be used for functional screening of mechanosensitive genes in vascular ECs, thus providing potential therapeutic targets to prevent or treat endothelial injury at atheroprone sites.
Issue Date: 10-Nov-2016
Date of Acceptance: 23-Oct-2016
URI: http://hdl.handle.net/10044/1/42976
DOI: http://dx.doi.org/10.1161/ATVBAHA.116.308502
ISSN: 1524-4636
Publisher: American Heart Association
Journal / Book Title: Arteriosclerosis Thrombosis and Vascular Biology
Volume: 36
Issue: 12
Copyright Statement: © 2016 American Heart Association, Inc.
Sponsor/Funder: British Heart Foundation
British Heart Foundation
Funder's Grant Number: RG/11/13/29055
Keywords: Zebrafish
endothelial cells
Cardiovascular System & Hematology
1103 Clinical Sciences
1102 Cardiovascular Medicine And Haematology
Publication Status: Published
Conference Place: United States
Appears in Collections:Bioengineering
National Heart and Lung Institute
Faculty of Engineering