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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...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2017
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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 |
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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 |
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