Comprehensive assessment of NR ligand polypharmacology by a multiplex reporter NR assay
Abstract Nuclear receptors (NR) are ligand-modulated transcription factors that regulate multiple cell functions and thus represent excellent drug targets. However, due to a considerable NR structural homology, NR ligands often interact with multiple receptors. Here, we describe a multiplex reporter...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2022-02-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-07031-8 |
| _version_ | 1818491739802959872 |
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| author | Alexander Medvedev Matt Moeser Liubov Medvedeva Elena Martsen Alexander Granick Lydia Raines Kristen Gorman Benjamin Lin Ming Zeng Keith A. Houck Sergei S. Makarov |
| author_facet | Alexander Medvedev Matt Moeser Liubov Medvedeva Elena Martsen Alexander Granick Lydia Raines Kristen Gorman Benjamin Lin Ming Zeng Keith A. Houck Sergei S. Makarov |
| author_sort | Alexander Medvedev |
| collection | DOAJ |
| description | Abstract Nuclear receptors (NR) are ligand-modulated transcription factors that regulate multiple cell functions and thus represent excellent drug targets. However, due to a considerable NR structural homology, NR ligands often interact with multiple receptors. Here, we describe a multiplex reporter assay (the FACTORIAL NR) that enables parallel assessment of NR ligand activity across all 48 human NRs. The assay comprises one-hybrid GAL4-NR reporter modules transiently transfected into test cells. To evaluate the reporter activity, we assessed their RNA transcripts. We used a homogeneous RNA detection approach that afforded equal detection efficacy and permitted the multiplex detection in a single-well format. For validation, we examined a panel of selective NR ligands and polypharmacological agonists and antagonists of the progestin, estrogen, PPAR, ERR, and ROR receptors. The assay produced highly reproducible NR activity profiles (r > 0.96) permitting quantitative assessment of individual NR responses. The inferred EC50 values agreed with the published data. The assay showed excellent quality (<Z’> = 0.73) and low variability (<CV> = 7.2%). Furthermore, the assay permitted distinguishing direct and non-direct NR responses to ligands. Therefore, the FACTORIAL NR enables comprehensive evaluation of NR ligand polypharmacology. |
| first_indexed | 2024-12-10T17:34:55Z |
| format | Article |
| id | doaj.art-7b4fcb73a0c54a51a995d71c913737bf |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-10T17:34:55Z |
| publishDate | 2022-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-7b4fcb73a0c54a51a995d71c913737bf2022-12-22T01:39:35ZengNature PortfolioScientific Reports2045-23222022-02-0112111810.1038/s41598-022-07031-8Comprehensive assessment of NR ligand polypharmacology by a multiplex reporter NR assayAlexander Medvedev0Matt Moeser1Liubov Medvedeva2Elena Martsen3Alexander Granick4Lydia Raines5Kristen Gorman6Benjamin Lin7Ming Zeng8Keith A. Houck9Sergei S. Makarov10Attagene IncAttagene IncAttagene IncAttagene IncAttagene IncAttagene IncAttagene IncAttagene IncAttagene IncUS Environmental Protection AgencyAttagene IncAbstract Nuclear receptors (NR) are ligand-modulated transcription factors that regulate multiple cell functions and thus represent excellent drug targets. However, due to a considerable NR structural homology, NR ligands often interact with multiple receptors. Here, we describe a multiplex reporter assay (the FACTORIAL NR) that enables parallel assessment of NR ligand activity across all 48 human NRs. The assay comprises one-hybrid GAL4-NR reporter modules transiently transfected into test cells. To evaluate the reporter activity, we assessed their RNA transcripts. We used a homogeneous RNA detection approach that afforded equal detection efficacy and permitted the multiplex detection in a single-well format. For validation, we examined a panel of selective NR ligands and polypharmacological agonists and antagonists of the progestin, estrogen, PPAR, ERR, and ROR receptors. The assay produced highly reproducible NR activity profiles (r > 0.96) permitting quantitative assessment of individual NR responses. The inferred EC50 values agreed with the published data. The assay showed excellent quality (<Z’> = 0.73) and low variability (<CV> = 7.2%). Furthermore, the assay permitted distinguishing direct and non-direct NR responses to ligands. Therefore, the FACTORIAL NR enables comprehensive evaluation of NR ligand polypharmacology.https://doi.org/10.1038/s41598-022-07031-8 |
| spellingShingle | Alexander Medvedev Matt Moeser Liubov Medvedeva Elena Martsen Alexander Granick Lydia Raines Kristen Gorman Benjamin Lin Ming Zeng Keith A. Houck Sergei S. Makarov Comprehensive assessment of NR ligand polypharmacology by a multiplex reporter NR assay Scientific Reports |
| title | Comprehensive assessment of NR ligand polypharmacology by a multiplex reporter NR assay |
| title_full | Comprehensive assessment of NR ligand polypharmacology by a multiplex reporter NR assay |
| title_fullStr | Comprehensive assessment of NR ligand polypharmacology by a multiplex reporter NR assay |
| title_full_unstemmed | Comprehensive assessment of NR ligand polypharmacology by a multiplex reporter NR assay |
| title_short | Comprehensive assessment of NR ligand polypharmacology by a multiplex reporter NR assay |
| title_sort | comprehensive assessment of nr ligand polypharmacology by a multiplex reporter nr assay |
| url | https://doi.org/10.1038/s41598-022-07031-8 |
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