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Isolation of NELL 1 Aptamers for Rhabdomyosarcoma Targeting

NELL1 (Neural epidermal growth factor-like (EGFL)-like protein) is an important biomarker associated with tissue and bone development and regeneration. NELL1 upregulation has been linked with metastasis and negative prognosis in rhabdomyosarcoma (RMS). Furthermore, multiple recent studies have also...

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Autores principales: Duan, Chengchen, Townley, Helen Elizabeth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9032205/
https://www.ncbi.nlm.nih.gov/pubmed/35447734
http://dx.doi.org/10.3390/bioengineering9040174
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author Duan, Chengchen
Townley, Helen Elizabeth
author_facet Duan, Chengchen
Townley, Helen Elizabeth
author_sort Duan, Chengchen
collection PubMed
description NELL1 (Neural epidermal growth factor-like (EGFL)-like protein) is an important biomarker associated with tissue and bone development and regeneration. NELL1 upregulation has been linked with metastasis and negative prognosis in rhabdomyosarcoma (RMS). Furthermore, multiple recent studies have also shown the importance of NELL1 in inflammatory bowel disease and membranous nephropathy, amongst other diseases. In this study, several anti-NELL1 DNA aptamers were selected from a randomized ssDNA pool using a fluorescence-guided method and evaluated for their binding affinity and selectivity. Several other methods such as a metabolic assay and confocal microscopy were also applied for the evaluation of the selected aptamers. The top three candidates were evaluated further, and AptNCan3 was shown to have a binding affinity up to 959.2 nM. Selectivity was examined in the RH30 RMS cells that overexpressed NELL1. Both AptNCan2 and AptNCan3 could significantly suppress metabolic activity in RMS cells. AptNCan3 was found to locate on the cell membrane and also on intracellular vesicles, which matched the location of NELL1 shown by antibodies in previous research. These results indicate that the selected anti-NELL1 aptamer showed strong and highly specific binding to NELL1 and therefore has potential to be used for in vitro or in vivo studies and treatments.
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spelling pubmed-90322052022-04-23 Isolation of NELL 1 Aptamers for Rhabdomyosarcoma Targeting Duan, Chengchen Townley, Helen Elizabeth Bioengineering (Basel) Article NELL1 (Neural epidermal growth factor-like (EGFL)-like protein) is an important biomarker associated with tissue and bone development and regeneration. NELL1 upregulation has been linked with metastasis and negative prognosis in rhabdomyosarcoma (RMS). Furthermore, multiple recent studies have also shown the importance of NELL1 in inflammatory bowel disease and membranous nephropathy, amongst other diseases. In this study, several anti-NELL1 DNA aptamers were selected from a randomized ssDNA pool using a fluorescence-guided method and evaluated for their binding affinity and selectivity. Several other methods such as a metabolic assay and confocal microscopy were also applied for the evaluation of the selected aptamers. The top three candidates were evaluated further, and AptNCan3 was shown to have a binding affinity up to 959.2 nM. Selectivity was examined in the RH30 RMS cells that overexpressed NELL1. Both AptNCan2 and AptNCan3 could significantly suppress metabolic activity in RMS cells. AptNCan3 was found to locate on the cell membrane and also on intracellular vesicles, which matched the location of NELL1 shown by antibodies in previous research. These results indicate that the selected anti-NELL1 aptamer showed strong and highly specific binding to NELL1 and therefore has potential to be used for in vitro or in vivo studies and treatments. MDPI 2022-04-15 /pmc/articles/PMC9032205/ /pubmed/35447734 http://dx.doi.org/10.3390/bioengineering9040174 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Duan, Chengchen
Townley, Helen Elizabeth
Isolation of NELL 1 Aptamers for Rhabdomyosarcoma Targeting
title Isolation of NELL 1 Aptamers for Rhabdomyosarcoma Targeting
title_full Isolation of NELL 1 Aptamers for Rhabdomyosarcoma Targeting
title_fullStr Isolation of NELL 1 Aptamers for Rhabdomyosarcoma Targeting
title_full_unstemmed Isolation of NELL 1 Aptamers for Rhabdomyosarcoma Targeting
title_short Isolation of NELL 1 Aptamers for Rhabdomyosarcoma Targeting
title_sort isolation of nell 1 aptamers for rhabdomyosarcoma targeting
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9032205/
https://www.ncbi.nlm.nih.gov/pubmed/35447734
http://dx.doi.org/10.3390/bioengineering9040174
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