Cargando…

Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent

Heparanase is overexpressed by tumor cells and degrades the extracellular matrix proteoglycans through cleavage of heparan sulfates (HS), allowing pro-angiogenic factor release and thus playing a key role in tumor angiogenesis and metastasis. Here we propose new HS analogs as potent heparanase inhib...

Descripción completa

Detalles Bibliográficos
Autores principales: Poupard, Nicolas, Badarou, Pamela, Fasani, Fabienne, Groult, Hugo, Bridiau, Nicolas, Sannier, Frédéric, Bordenave-Juchereau, Stéphanie, Kieda, Claudine, Piot, Jean-Marie, Grillon, Catherine, Fruitier-Arnaudin, Ingrid, Maugard, Thierry
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5450540/
https://www.ncbi.nlm.nih.gov/pubmed/28486399
http://dx.doi.org/10.3390/md15050134
_version_ 1783239999900090368
author Poupard, Nicolas
Badarou, Pamela
Fasani, Fabienne
Groult, Hugo
Bridiau, Nicolas
Sannier, Frédéric
Bordenave-Juchereau, Stéphanie
Kieda, Claudine
Piot, Jean-Marie
Grillon, Catherine
Fruitier-Arnaudin, Ingrid
Maugard, Thierry
author_facet Poupard, Nicolas
Badarou, Pamela
Fasani, Fabienne
Groult, Hugo
Bridiau, Nicolas
Sannier, Frédéric
Bordenave-Juchereau, Stéphanie
Kieda, Claudine
Piot, Jean-Marie
Grillon, Catherine
Fruitier-Arnaudin, Ingrid
Maugard, Thierry
author_sort Poupard, Nicolas
collection PubMed
description Heparanase is overexpressed by tumor cells and degrades the extracellular matrix proteoglycans through cleavage of heparan sulfates (HS), allowing pro-angiogenic factor release and thus playing a key role in tumor angiogenesis and metastasis. Here we propose new HS analogs as potent heparanase inhibitors: Heparin as a positive control, Dextran Sulfate, λ-Carrageenan, and modified forms of them obtained by depolymerization associated to glycol splitting (RD-GS). After heparanase activity assessment, 11 kDa RD-GS-λ-Carrageenan emerged as the most effective heparanase inhibitor with an IC(50) of 7.32 ng/mL compared to 10.7 ng/mL for the 16 kDa unfractionated heparin. The fractionated polysaccharides were then tested in a heparanase-rich medium-based in vitro model, mimicking tumor microenvironment, to determine their effect on microvascular endothelial cells (HSkMEC) angiogenesis. As a preliminary study, we identified that under hypoxic and nutrient poor conditions, MCF-7 cancer cells released much more mature heparanase in their supernatant than in normal conditions. Then a Matrigel(TM) assay using HSkMEC cultured under hypoxic conditions in the presence (or not) of this heparanase-rich supernatant was realized. Adding heparanase-rich media strongly enhanced angiogenic network formation with a production of twice more pseudo-vessels than with the control. When sulfated polysaccharides were tested in this angiogenesis assay, RD-GS-λ-Carrageenan was identified as a promising anti-angiogenic agent.
format Online
Article
Text
id pubmed-5450540
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-54505402017-06-05 Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent Poupard, Nicolas Badarou, Pamela Fasani, Fabienne Groult, Hugo Bridiau, Nicolas Sannier, Frédéric Bordenave-Juchereau, Stéphanie Kieda, Claudine Piot, Jean-Marie Grillon, Catherine Fruitier-Arnaudin, Ingrid Maugard, Thierry Mar Drugs Article Heparanase is overexpressed by tumor cells and degrades the extracellular matrix proteoglycans through cleavage of heparan sulfates (HS), allowing pro-angiogenic factor release and thus playing a key role in tumor angiogenesis and metastasis. Here we propose new HS analogs as potent heparanase inhibitors: Heparin as a positive control, Dextran Sulfate, λ-Carrageenan, and modified forms of them obtained by depolymerization associated to glycol splitting (RD-GS). After heparanase activity assessment, 11 kDa RD-GS-λ-Carrageenan emerged as the most effective heparanase inhibitor with an IC(50) of 7.32 ng/mL compared to 10.7 ng/mL for the 16 kDa unfractionated heparin. The fractionated polysaccharides were then tested in a heparanase-rich medium-based in vitro model, mimicking tumor microenvironment, to determine their effect on microvascular endothelial cells (HSkMEC) angiogenesis. As a preliminary study, we identified that under hypoxic and nutrient poor conditions, MCF-7 cancer cells released much more mature heparanase in their supernatant than in normal conditions. Then a Matrigel(TM) assay using HSkMEC cultured under hypoxic conditions in the presence (or not) of this heparanase-rich supernatant was realized. Adding heparanase-rich media strongly enhanced angiogenic network formation with a production of twice more pseudo-vessels than with the control. When sulfated polysaccharides were tested in this angiogenesis assay, RD-GS-λ-Carrageenan was identified as a promising anti-angiogenic agent. MDPI 2017-05-09 /pmc/articles/PMC5450540/ /pubmed/28486399 http://dx.doi.org/10.3390/md15050134 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Poupard, Nicolas
Badarou, Pamela
Fasani, Fabienne
Groult, Hugo
Bridiau, Nicolas
Sannier, Frédéric
Bordenave-Juchereau, Stéphanie
Kieda, Claudine
Piot, Jean-Marie
Grillon, Catherine
Fruitier-Arnaudin, Ingrid
Maugard, Thierry
Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent
title Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent
title_full Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent
title_fullStr Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent
title_full_unstemmed Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent
title_short Assessment of Heparanase-Mediated Angiogenesis Using Microvascular Endothelial Cells: Identification of λ-Carrageenan Derivative as a Potent Anti Angiogenic Agent
title_sort assessment of heparanase-mediated angiogenesis using microvascular endothelial cells: identification of λ-carrageenan derivative as a potent anti angiogenic agent
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5450540/
https://www.ncbi.nlm.nih.gov/pubmed/28486399
http://dx.doi.org/10.3390/md15050134
work_keys_str_mv AT poupardnicolas assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT badaroupamela assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT fasanifabienne assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT groulthugo assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT bridiaunicolas assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT sannierfrederic assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT bordenavejuchereaustephanie assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT kiedaclaudine assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT piotjeanmarie assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT grilloncatherine assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT fruitierarnaudiningrid assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent
AT maugardthierry assessmentofheparanasemediatedangiogenesisusingmicrovascularendothelialcellsidentificationoflcarrageenanderivativeasapotentantiangiogenicagent