NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells
Increasing evidence has revealed the role of G protein-coupled receptors (GPCRs) in the regulation of signalling responses via complex cross-talk mechanisms with the receptor tyrosine kinase (RTK) family of transmembrane receptors. Vascular endothelial growth factor-a (VEGFa) mediates cancer angioge...
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| Format: | Thesis (University of Nottingham only) |
| Language: | English |
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2019
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| Online Access: | https://eprints.nottingham.ac.uk/56087/ |
| _version_ | 1848799269572575232 |
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| author | Carvalheira Alcobia, Diana |
| author_facet | Carvalheira Alcobia, Diana |
| author_sort | Carvalheira Alcobia, Diana |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Increasing evidence has revealed the role of G protein-coupled receptors (GPCRs) in the regulation of signalling responses via complex cross-talk mechanisms with the receptor tyrosine kinase (RTK) family of transmembrane receptors. Vascular endothelial growth factor-a (VEGFa) mediates cancer angiogenesis via binding to its cognate VEGF receptor 2 (VEGFR2). The two class A GPCRs, adenosine A2A receptors and β2-adrenoceptors, have also been intimately involved in cancer angiogenesis, and their activation can contribute to cancer progression and invasion. However, the molecular mechanisms involved in the cross-talk between these receptor families are not well understood.
Using a NanoLuc-based bioluminescence resonance energy transfer (NanoBRET) methodology, we reveal a novel mechanism in which VEGFR2 can associate with either of these Gs-coupled GPCRs to form oligomeric complexes in living cells. We also demonstrated that VEGF-stimulated VEGFR2 can induce a switch from a transient to a more stable interaction between β2AR and the adaptor protein β-arrestin-2, which may have an impact in endosomal signalling.
The pharmacological inhibition of β2-adrenoceptor (β2AR) signalling, using selective β2AR antagonists (or β-blockers), represents a potential therapeutic approach for the treatment of triple-negative breast cancer variant, which has limited treatment options. This study demonstrated the novel application of NanoBRET technology to probe specific β-blocker-β2-adrenoceptor engagement in a pre-clinical in vivo model of triple-negative breast cancer. This novel methodology will allow a strong correlation between drug-target engagement and mediated physiological response. |
| first_indexed | 2025-11-14T20:32:59Z |
| format | Thesis (University of Nottingham only) |
| id | nottingham-56087 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T20:32:59Z |
| publishDate | 2019 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-560872025-02-28T14:23:59Z https://eprints.nottingham.ac.uk/56087/ NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells Carvalheira Alcobia, Diana Increasing evidence has revealed the role of G protein-coupled receptors (GPCRs) in the regulation of signalling responses via complex cross-talk mechanisms with the receptor tyrosine kinase (RTK) family of transmembrane receptors. Vascular endothelial growth factor-a (VEGFa) mediates cancer angiogenesis via binding to its cognate VEGF receptor 2 (VEGFR2). The two class A GPCRs, adenosine A2A receptors and β2-adrenoceptors, have also been intimately involved in cancer angiogenesis, and their activation can contribute to cancer progression and invasion. However, the molecular mechanisms involved in the cross-talk between these receptor families are not well understood. Using a NanoLuc-based bioluminescence resonance energy transfer (NanoBRET) methodology, we reveal a novel mechanism in which VEGFR2 can associate with either of these Gs-coupled GPCRs to form oligomeric complexes in living cells. We also demonstrated that VEGF-stimulated VEGFR2 can induce a switch from a transient to a more stable interaction between β2AR and the adaptor protein β-arrestin-2, which may have an impact in endosomal signalling. The pharmacological inhibition of β2-adrenoceptor (β2AR) signalling, using selective β2AR antagonists (or β-blockers), represents a potential therapeutic approach for the treatment of triple-negative breast cancer variant, which has limited treatment options. This study demonstrated the novel application of NanoBRET technology to probe specific β-blocker-β2-adrenoceptor engagement in a pre-clinical in vivo model of triple-negative breast cancer. This novel methodology will allow a strong correlation between drug-target engagement and mediated physiological response. 2019-07-19 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/56087/1/Thesis%20final_after%20viva%20corrections_PDF.pdf Carvalheira Alcobia, Diana (2019) NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells. PhD thesis, University of Nottingham. NanoBRET Pharmacology Breast cancer |
| spellingShingle | NanoBRET Pharmacology Breast cancer Carvalheira Alcobia, Diana NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells |
| title | NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells |
| title_full | NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells |
| title_fullStr | NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells |
| title_full_unstemmed | NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells |
| title_short | NanoBRET to probe ligand-receptor and receptor-receptor interactions in living cells |
| title_sort | nanobret to probe ligand-receptor and receptor-receptor interactions in living cells |
| topic | NanoBRET Pharmacology Breast cancer |
| url | https://eprints.nottingham.ac.uk/56087/ |