Synthetic FXR agonist GW4064 is a modulator of multiple G protein-coupled receptors

Nidhi Singh, Manisha Yadav, Abhishek Kumar Singh, Harish Kumar, Shailendra Kumar Dhar Dwivedi, Jay Sharan Mishra, Anagha Gurjar, Amit Manhas, Sharat Chandra, Prem Narayan Yadav, Kumaravelu Jagavelu, Mohammad Imran Siddiqi, Arun Kumar Trivedi, Naibedya Chattopadhyay, Sabyasachi Sanyal

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The synthetic nuclear bile acid receptor (farnesoid X receptor [FXR]) agonist GW4064 is extensively used as a specific pharmacological tool to illustrate FXR functions. We noticed that GW4064 activated empty luciferase reporters in FXR-deficient HEK-293T cells.Wepostulated that this activity of GW4064 might be routed through as yet unknown cellular targets and undertook an unbiased exploratory approach to identify these targets. Investigations revealed that GW4064 activated cAMP and nuclear factor for activated T-cell response elements (CRE and NFAT-RE, respectively) present on these empty reporters. Whereas GW4064-induced NFAT-RE activation involved rapid intracellular Ca2+ accumulation and NFAT nuclear translocation, CRE activation involved soluble adenylyl cyclase-dependent cAMP accumulation and Ca2+-calcineurin-dependent nuclear translocation of transducers of regulated CRE-binding protein 2. Use of dominant negative heterotrimeric G-protein minigenes revealed that GW4064 caused activation of Gαi/o and Gq/11 G proteins. Sequential pharmacological inhibitor- based screening and radioligand-binding studies revealed that GW4064 interacted with multiple G protein-coupled receptors. Functional studies demonstrated that GW4064 robustly activated H1 and H4and inhibitedH2histamine receptor signaling events.Wealso found that MCF-7 breast cancer cells, reported to undergo GW4064-induced apoptosis in an FXR-dependent manner, did not express FXR, and the GW4064-mediated apoptosis, also apparent in HEK-293T cells, could be blocked by selective histamine receptor regulators. Taken together, our results demonstrate identification of histamine receptors as alternate targets for GW4064, which not only necessitates cautious interpretation of the biological functions attributed to FXR using GW4064 as a pharmacological tool but also provides a basis for the rational designing of new pharmacophores for histamine receptor modulation.

Original languageEnglish (US)
Pages (from-to)659-673
Number of pages15
JournalMolecular Endocrinology
Volume28
Issue number5
DOIs
StatePublished - 2014
Externally publishedYes

Fingerprint

G-Protein-Coupled Receptors
Histamine Receptors
HEK293 Cells
Pharmacology
GW 4064
Gq-G11 GTP-Binding Protein alpha Subunits
Apoptosis
NFATC Transcription Factors
Heterotrimeric GTP-Binding Proteins
Calcineurin
Response Elements
Luciferases
Transducers
Bile Acids and Salts
Adenylyl Cyclases
Carrier Proteins
Breast Neoplasms

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology
  • Medicine(all)

Cite this

Singh, N., Yadav, M., Singh, A. K., Kumar, H., Dhar Dwivedi, S. K., Mishra, J. S., ... Sanyal, S. (2014). Synthetic FXR agonist GW4064 is a modulator of multiple G protein-coupled receptors. Molecular Endocrinology, 28(5), 659-673. https://doi.org/10.1210/me.2013-1353

Synthetic FXR agonist GW4064 is a modulator of multiple G protein-coupled receptors. / Singh, Nidhi; Yadav, Manisha; Singh, Abhishek Kumar; Kumar, Harish; Dhar Dwivedi, Shailendra Kumar; Mishra, Jay Sharan; Gurjar, Anagha; Manhas, Amit; Chandra, Sharat; Yadav, Prem Narayan; Jagavelu, Kumaravelu; Siddiqi, Mohammad Imran; Trivedi, Arun Kumar; Chattopadhyay, Naibedya; Sanyal, Sabyasachi.

In: Molecular Endocrinology, Vol. 28, No. 5, 2014, p. 659-673.

Research output: Contribution to journalArticle

Singh, N, Yadav, M, Singh, AK, Kumar, H, Dhar Dwivedi, SK, Mishra, JS, Gurjar, A, Manhas, A, Chandra, S, Yadav, PN, Jagavelu, K, Siddiqi, MI, Trivedi, AK, Chattopadhyay, N & Sanyal, S 2014, 'Synthetic FXR agonist GW4064 is a modulator of multiple G protein-coupled receptors', Molecular Endocrinology, vol. 28, no. 5, pp. 659-673. https://doi.org/10.1210/me.2013-1353
Singh N, Yadav M, Singh AK, Kumar H, Dhar Dwivedi SK, Mishra JS et al. Synthetic FXR agonist GW4064 is a modulator of multiple G protein-coupled receptors. Molecular Endocrinology. 2014;28(5):659-673. https://doi.org/10.1210/me.2013-1353
Singh, Nidhi ; Yadav, Manisha ; Singh, Abhishek Kumar ; Kumar, Harish ; Dhar Dwivedi, Shailendra Kumar ; Mishra, Jay Sharan ; Gurjar, Anagha ; Manhas, Amit ; Chandra, Sharat ; Yadav, Prem Narayan ; Jagavelu, Kumaravelu ; Siddiqi, Mohammad Imran ; Trivedi, Arun Kumar ; Chattopadhyay, Naibedya ; Sanyal, Sabyasachi. / Synthetic FXR agonist GW4064 is a modulator of multiple G protein-coupled receptors. In: Molecular Endocrinology. 2014 ; Vol. 28, No. 5. pp. 659-673.
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