Development of new methods for determining the heparanase enzymatic activity

Melo, Carina Mucciolo; Tersariol, Ivarne L.S.; Nader, Helena B.; Pinhal, Maria Aparecida Silva; Lima, Marcelo A.

doi:10.1016/j.carres.2015.04.020

Cited by 18 publications

(14 citation statements)

References 43 publications

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“…20.1b) clinically used as antithrombotic agent [43]. As a good heparanase substrate, it has been included as a component of heparanase activity assay kit particularly useful for kinetic analysis and screening of enzyme inhibitors [44,45]. The heparanase cleavage site was described to be also dependent on the sulfation pattern of the neighboring sequences [46].…”

Section: Heparanase: Discovery and Characterizationmentioning

confidence: 99%

Non-Anticoagulant Heparins as Heparanase Inhibitors

Cassinelli

Torri

Naggi

2020

Advances in Experimental Medicine and Biology

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Section: Heparanase: Discovery and Characterizationmentioning

confidence: 99%

Non-Anticoagulant Heparins as Heparanase Inhibitors

Cassinelli

Torri

Naggi

2020

Advances in Experimental Medicine and Biology

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“…Human endo‐β‐glucuronidases, such as heparanases, are responsible for the hydrolysis of β‐D‐glucuronic acid residues from the non‐reducing end of heparan sulfates (McCarter & Withers, ; Melo, Tersariol, Nader, Pinhal, & Lima, ). Heparanase expression is increased in glioma specimens in tumor and endothelial cells, and its expression has been correlated with proliferation, migration (Hong et al, ; Hong, Nelson, deCarvalho, & Kalkanis, ) and angiogenesis (Kundu et al, ; Zetser, Bashenko, Miao, Vlodavsky, & Ilan, ); as seen in other types of tumors (Cassinelli, Zaffaroni, & Lanzi, ; Crispel et al, ; Elkin et al, ; Vlodavsky, Elkin, & Ilan, ; Vornicova et al, ).…”

Section: Modulation Of Brain Extracellular Matrix By Proteases and Glmentioning

confidence: 99%

Glioma infiltration and extracellular matrix: key players and modulators

Ferrer

Moura‐Neto

Mentlein

2018

Glia

166

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An outstanding characteristic of gliomas is their infiltration into brain parenchyma, a property that impairs complete surgical resection; consequently, these tumors might recur, resulting in a high mortality rate. Gliomas invade along preferential routes, such as those along white matter tracts and in the perineuronal and perivascular spaces. Brain extracellular components and their partners and modulators play a crucial role in glioma cell invasion. This review presents an extensive survey of the literature, showing how the brain extracellular matrix (ECM) is modulated during the glioma infiltration process. We explore aspects of ECM interaction with glioma cells, reviewing the main glycosaminoglycans, glycoproteins and proteoglycans. We discuss the roles of ECM-binding proteins, including CD44, RHAMM, integrins and axonal guidance molecules, and highlight the role of proteases and glycosidases in glioma infiltration; in binding and release chemokines, cytokines and growth factors; and in generating new bioactive ECM fragments. We also consider the roles of cytoskeletal signaling, angiogenesis, miRNAs and the glial-to-mesenchymal transition linked to glioma invasion. We closely discuss therapeutic approaches based on the modulation of the extracellular matrix, targeting the control of glioma infiltration, its relative failure in clinical trials, and potential means to overcome this difficulty.

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“…Recently, biotinylated HS was immobilised onto a protamine-coated plate. Following treatment with heparanase, the immobilised biotinylated HS not digested by heparanase was detected with a Eu-streptavidin conjugate [ 42 ]. Wu and coworkers [ 43 ] biotinylated HS at the non-reducing end in an elaborate but controlled fashion by first enzymatically attaching an azido-labelled sugar, GlcNAz, with the glycosyltransferase EXT1/2 to HS chains attached to the proteoglycan rhSynd4.…”

Section: Assaying Heparanasementioning

confidence: 99%

“…This assay is more rapid than the WST-1 method and is not affected by the presence of reducing sugars in the sample; however, it is unclear whether it can be adapted to biological samples because of the presence of multiple endogenous glycosaminoglycans (GAGs) in the latter. The second assay is a variation of the WST-1 assay and uses the sodium salt of resazurin [ 42 ], a commercially available dye which also reacts with reducing sugars. Unlike WST-1, the product of the reaction is fluorescent and is detected at 590 nm following excitation at 560 nm.…”

Section: Assaying Heparanasementioning

confidence: 99%

The Development of Assays for Heparanase Enzymatic Activity: Towards a Gold Standard

Chhabra

Ferro

2018

Molecules

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The enzyme heparanase, an endo-β-glucuronidase, degrades heparan sulfate (HS) chains on the cell surface and in the extracellular matrix. Heparanase regulates numerous biological processes that drive tumour growth, metastasis and angiogenesis. In addition to its key role in cancer progression, it has also been implicated in an ever-growing number of other diseases, particularly those associated with inflammation. The importance of heparanase in biology has led to numerous efforts over the years to develop assays to monitor its activity and to screen for new inhibitors as potential drug candidates. Despite these efforts and the commercialization of a few kits, most heparanase assays are still complex, labour intensive, costly or have limited application. Herein we review the various methods for assaying heparanase enzymatic activity, focusing on recent developments towards new assays that hold the promise of accelerating research into this important enzyme.

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Development of new methods for determining the heparanase enzymatic activity

Cited by 18 publications

References 43 publications

Non-Anticoagulant Heparins as Heparanase Inhibitors

Non-Anticoagulant Heparins as Heparanase Inhibitors

Glioma infiltration and extracellular matrix: key players and modulators

The Development of Assays for Heparanase Enzymatic Activity: Towards a Gold Standard

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