A drug-repositioning screen using splicing-sensitive fluorescent reporters identifies novel modulators of VEGF-A splicing with anti-angiogenic properties
dc.contributor.author | Star, E | |
dc.contributor.author | Stevens, M | |
dc.contributor.author | Gooding, C | |
dc.contributor.author | Smith, CWJ | |
dc.contributor.author | Li, L | |
dc.contributor.author | Ayine, ML | |
dc.contributor.author | Harper, SJ | |
dc.contributor.author | Bates, DO | |
dc.contributor.author | Oltean, S | |
dc.date.accessioned | 2021-05-06T09:50:19Z | |
dc.date.issued | 2021-04-09 | |
dc.description.abstract | Alternative splicing of the vascular endothelial growth factor A (VEGF-A) terminal exon generates two protein families with differing functions. Pro-angiogenic VEGF-Axxxa isoforms are produced via selection of the proximal 3′ splice site of the terminal exon. Use of an alternative distal splice site generates the anti-angiogenic VEGF-Axxxb proteins. A bichromatic splicing-sensitive reporter was designed to mimic VEGF-A alternative splicing and was used as a molecular tool to further investigate this alternative splicing event. Part of VEGF-A’s terminal exon and preceding intron were inserted into a minigene construct followed by the coding sequences for two fluorescent proteins. A different fluorescent protein is expressed depending on which 3′ splice site of the exon is used during splicing (dsRED denotes VEGF-Axxxa and EGFP denotes VEGF-Axxxb). The fluorescent output can be used to follow splicing decisions in vitro and in vivo. Following successful reporter validation in different cell lines and altering splicing using known modulators, a screen was performed using the LOPAC library of small molecules. Alterations to reporter splicing were measured using a fluorescent plate reader to detect dsRED and EGFP expression. Compounds of interest were further validated using flow cytometry and assessed for effects on endogenous VEGF-A alternative splicing at the mRNA and protein level. Ex vivo and in vitro angiogenesis assays were used to demonstrate the anti-angiogenic effect of the compounds. Furthermore, anti-angiogenic activity was investigated in a Matrigel in vivo model. To conclude, we have identified a set of compounds that have anti-angiogenic activity through modulation of VEGF-A terminal exon splicing. | en_GB |
dc.description.sponsorship | Biotechnology and Biological Sciences Research Council (BBSRC) | en_GB |
dc.description.sponsorship | British Heart Foundation | en_GB |
dc.description.sponsorship | Diabetes UK | en_GB |
dc.identifier.citation | Vol. 10, article 36 | en_GB |
dc.identifier.doi | 10.1038/s41389-021-00323-0 | |
dc.identifier.grantnumber | BB/J007293/2 | en_GB |
dc.identifier.grantnumber | PG/15/53/31371 | en_GB |
dc.identifier.grantnumber | 17/0005668 | en_GB |
dc.identifier.uri | http://hdl.handle.net/10871/125571 | |
dc.language.iso | en | en_GB |
dc.publisher | Springer Nature | en_GB |
dc.rights | © The Author(s) 2021. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. | en_GB |
dc.title | A drug-repositioning screen using splicing-sensitive fluorescent reporters identifies novel modulators of VEGF-A splicing with anti-angiogenic properties | en_GB |
dc.type | Article | en_GB |
dc.date.available | 2021-05-06T09:50:19Z | |
exeter.article-number | 36 | en_GB |
dc.description | This is the final version. Available on open access from Springer Nature via the DOI in this record | en_GB |
dc.identifier.eissn | 2157-9024 | |
dc.identifier.journal | Oncogenesis | en_GB |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | en_GB |
dcterms.dateAccepted | 2021-03-25 | |
exeter.funder | ::British Heart Foundation | en_GB |
exeter.funder | ::Diabetes UK | en_GB |
rioxxterms.version | VoR | en_GB |
rioxxterms.licenseref.startdate | 2021-04-09 | |
rioxxterms.type | Journal Article/Review | en_GB |
refterms.dateFCD | 2021-05-06T09:43:34Z | |
refterms.versionFCD | VoR | |
refterms.dateFOA | 2021-05-06T09:50:38Z | |
refterms.panel | A | en_GB |
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Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction
in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if
changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If
material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.