Selective Targeting to Glioma with Nucleic Acid Aptamers

Malignant glioma is characterised by a rapid growth rate and high capacity for invasive infiltration to surrounding brain tissue; hence, diagnosis and treatment is difficult and patient survival is poor. Aptamers contribute a promising and unique technology for the in vitro imaging of live cells and...

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Autores principales: Aptekar, Shraddha, Arora, Mohit, Lawrence, Clare Louise, Lea, Robert William, Ashton, Katherine, Dawson, Tim, Alder, Jane Elizabeth, Shaw, Lisa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4529171/
https://www.ncbi.nlm.nih.gov/pubmed/26252900
http://dx.doi.org/10.1371/journal.pone.0134957
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author Aptekar, Shraddha
Arora, Mohit
Lawrence, Clare Louise
Lea, Robert William
Ashton, Katherine
Dawson, Tim
Alder, Jane Elizabeth
Shaw, Lisa
author_facet Aptekar, Shraddha
Arora, Mohit
Lawrence, Clare Louise
Lea, Robert William
Ashton, Katherine
Dawson, Tim
Alder, Jane Elizabeth
Shaw, Lisa
author_sort Aptekar, Shraddha
collection PubMed
description Malignant glioma is characterised by a rapid growth rate and high capacity for invasive infiltration to surrounding brain tissue; hence, diagnosis and treatment is difficult and patient survival is poor. Aptamers contribute a promising and unique technology for the in vitro imaging of live cells and tissues, with a potentially bright future in clinical diagnostics and therapeutics for malignant glioma. The binding selectivity, uptake capacity and binding target of two DNA aptamers, SA43 and SA44, were investigated in glioma cells and patient tissues. The binding assay showed that SA43 and SA44 bound with strong affinity (K(d), 21.56 ± 4.60 nM and K(d), 21.11 ± 3.30 nM respectively) to the target U87MG cells. Quantitative analysis by flow cytometry showed that the aptamers were able to actively internalise in U87MG and 1321N1 glioma cells compared to the non-cancerous and non-glioma cell types. Confocal microscopy confirmed staining in the cytoplasm, and co-localisation studies with endoplasmic reticulum, Golgi apparatus and lysosomal markers suggested internalisation and compartmentalisation within the endomembrane system. Both aptamers selectively bound to Ku 70 and Ku 80 DNA repair proteins as determined by aptoprecipitation (AP) followed by mass spectrometry analysis and confirmation by Western blot. In addition, aptohistochemical (AHC) staining on paraffin embedded, formalin fixed patient tissues revealed that the binding selectivity was significantly higher for SA43 aptamer in glioma tissues (grade I, II, III and IV) compared to the non-cancerous tissues, whereas SA44 did not show selectivity towards glioma tissues. The results indicate that SA43 aptamer can differentiate between glioma and non-cancerous cells and tissues and therefore, shows promise for histological diagnosis of glioma.
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spelling pubmed-45291712015-08-12 Selective Targeting to Glioma with Nucleic Acid Aptamers Aptekar, Shraddha Arora, Mohit Lawrence, Clare Louise Lea, Robert William Ashton, Katherine Dawson, Tim Alder, Jane Elizabeth Shaw, Lisa PLoS One Research Article Malignant glioma is characterised by a rapid growth rate and high capacity for invasive infiltration to surrounding brain tissue; hence, diagnosis and treatment is difficult and patient survival is poor. Aptamers contribute a promising and unique technology for the in vitro imaging of live cells and tissues, with a potentially bright future in clinical diagnostics and therapeutics for malignant glioma. The binding selectivity, uptake capacity and binding target of two DNA aptamers, SA43 and SA44, were investigated in glioma cells and patient tissues. The binding assay showed that SA43 and SA44 bound with strong affinity (K(d), 21.56 ± 4.60 nM and K(d), 21.11 ± 3.30 nM respectively) to the target U87MG cells. Quantitative analysis by flow cytometry showed that the aptamers were able to actively internalise in U87MG and 1321N1 glioma cells compared to the non-cancerous and non-glioma cell types. Confocal microscopy confirmed staining in the cytoplasm, and co-localisation studies with endoplasmic reticulum, Golgi apparatus and lysosomal markers suggested internalisation and compartmentalisation within the endomembrane system. Both aptamers selectively bound to Ku 70 and Ku 80 DNA repair proteins as determined by aptoprecipitation (AP) followed by mass spectrometry analysis and confirmation by Western blot. In addition, aptohistochemical (AHC) staining on paraffin embedded, formalin fixed patient tissues revealed that the binding selectivity was significantly higher for SA43 aptamer in glioma tissues (grade I, II, III and IV) compared to the non-cancerous tissues, whereas SA44 did not show selectivity towards glioma tissues. The results indicate that SA43 aptamer can differentiate between glioma and non-cancerous cells and tissues and therefore, shows promise for histological diagnosis of glioma. Public Library of Science 2015-08-07 /pmc/articles/PMC4529171/ /pubmed/26252900 http://dx.doi.org/10.1371/journal.pone.0134957 Text en © 2015 Aptekar et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Aptekar, Shraddha
Arora, Mohit
Lawrence, Clare Louise
Lea, Robert William
Ashton, Katherine
Dawson, Tim
Alder, Jane Elizabeth
Shaw, Lisa
Selective Targeting to Glioma with Nucleic Acid Aptamers
title Selective Targeting to Glioma with Nucleic Acid Aptamers
title_full Selective Targeting to Glioma with Nucleic Acid Aptamers
title_fullStr Selective Targeting to Glioma with Nucleic Acid Aptamers
title_full_unstemmed Selective Targeting to Glioma with Nucleic Acid Aptamers
title_short Selective Targeting to Glioma with Nucleic Acid Aptamers
title_sort selective targeting to glioma with nucleic acid aptamers
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4529171/
https://www.ncbi.nlm.nih.gov/pubmed/26252900
http://dx.doi.org/10.1371/journal.pone.0134957
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