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Development of novel single-stranded nucleic acid aptamers against the pro-Angiogenic and metastatic enzyme heparanase (HPSE1)

Simmons, Suzanne; Velasco, Maria and Missailidis, Sotiris (2012). Development of novel single-stranded nucleic acid aptamers against the pro-Angiogenic and metastatic enzyme heparanase (HPSE1). PLoS ONE

URL: http://www.plosone.org/article/info%3Adoi%2F10.137...
DOI (Digital Object Identifier) Link: http://dx.doi.org/10.1371/journal.pone.0037938
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Abstract

Heparanase is an enzyme involved in extracellular matrix remodelling and heparan sulphate proteoglycan catabolism. It is secreted by metastatic tumour cells, allowing them to penetrate the endothelial cell layer and basement membrane to invade target organs. The release of growth factors at the site of cleaved heparan sulphate chains further enhance the potential of the tumour by encouraging the process of angiogenesis. This leads to increased survival and further proliferation of the tumour. Aptamers are single or double stranded oligonucleotides that recognise specific small molecules, peptides, proteins, or even cells or tissues and have shown great potential over the years as diagnostic and therapeutic agents in anticancer treatment. For the first time, single stranded DNA aptamers were successfully generated against the active heterodimer form of heparanase using a modified SELEX protocol, and eluted based on increasing affinity for the target. Sandwich ELISA assays showed recognition of heparanase by the aptamers at a site distinct from that of a polyclonal HPSE1 antibody. The binding affinities of aptamer to immobilised enzyme were high (7×107 to 8×107 M−1) as measured by fluorescence spectroscopy. Immunohistochemistry and immunofluorescence studies demonstrated that the aptamers were able to recognise heparanase with staining comparable or in some cases superior to that of the HPSE1 antibody control. Finally, matrigel assay demonstrated that aptamers were able to inhibit heparanase. This study provides clear proof of principle concept that nucleic acid aptamers can be generated against heparanase. These reagents may serve as useful tools to explore the functional role of the enzyme and in the future development of diagnostic assays or therapeutic reagents.

Item Type: Journal Article
Copyright Holders: 2012 Simmons et al
ISSN: 1932-6203
Project Funding Details:
Funded Project NameProject IDFunding Body
Not SetNot SetThe Open University
Academic Unit/Department: Science > Life, Health and Chemical Sciences
Interdisciplinary Research Centre: Centre for Earth, Planetary, Space and Astronomical Research (CEPSAR)
Biomedical Research Network (BRN)
Item ID: 33922
Depositing User: Maria Velasco
Date Deposited: 27 Jun 2012 12:07
Last Modified: 07 Mar 2014 15:00
URI: http://oro.open.ac.uk/id/eprint/33922
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