SpringerLink
Log in

Cell-Based Assays to Identify ERR and ERR/PGC Modulators

  • Protocol
  • First Online:
High-Throughput Screening Assays in Toxicology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2474))

Abstract

The estrogen-related receptor alpha (ERRα, NR3B1) is an orphan nuclear receptor which plays a role in endocrine disruption, energy homeostasis, and cancer prognosis. One of the unique features of this transcription factor is the interplay with its cofactors. For instance, certain modulators require the presence of proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) alongside ERRα. Therefore, identification of ERRα agonists and antagonists require examination of this nuclear receptor alone and together with PGC-1α. In this book chapter, we describe the step-by-step protocol of a multiplex luciferase assay designed to identify ERRα agonists, antagonists, and toxicity in one quantitative high-throughput screening assay using two different stable cell lines.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
GBP 34.95
Price includes VAT (United Kingdom)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
GBP 87.50
Price includes VAT (United Kingdom)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
GBP 109.99
Price includes VAT (United Kingdom)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info
Hardcover Book
GBP 139.99
Price includes VAT (United Kingdom)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Huss JM, Imahashi KI, Dufour CR, Weinheimer CJ, Courtois M, Kovacs A, Giguere V, Murphy E, Kelly DP (2007) The nuclear receptor ERR alpha is required for the bioenergetic and functional adaptation to cardiac pressure overload. Cell Metab 6(1):25–37. https://doi.org/10.1016/j.cmet.2007.06.005

    Article  CAS  PubMed  Google Scholar 

  2. Tripathi M, Yen PM, Singh BK (2020) Estrogen-related receptor alpha: an under-appreciated potential target for the treatment of metabolic diseases. Int J Mol Sci 21(5):1645. https://doi.org/10.3390/ijms21051645

    Article  CAS  PubMed Central  Google Scholar 

  3. Giguere V, Yang N, Segui P, Evans RM (1988) Identification of a new class of steroid-hormone receptors. Nature 331(6151):91–94. https://doi.org/10.1038/331091a0

    Article  CAS  PubMed  Google Scholar 

  4. Giguere V (2002) To ERR in the estrogen pathway. Trends Endocrinol Metab 13(5):220–225. https://doi.org/10.1016/s1043-2760(02)00592-1

    Article  CAS  PubMed  Google Scholar 

  5. Liu D, Zhang Z, Gladwell W, Teng CT (2003) Estrogen stimulates estrogen-related receptor alpha gene expression through conserved hormone response elements. Endocrinology 144(11):4894–4904. https://doi.org/10.1210/en.2003-0432

    Article  CAS  PubMed  Google Scholar 

  6. Ariazi EA, Clark GM, Mertz JE (2002) Estrogen-related receptor alpha and estrogen-related receptor gamma associate with unfavorable and favorable biomarkers, respectively, in human breast cancer. Cancer Res 62(22):6510–6518

    CAS  PubMed  Google Scholar 

  7. Ariazi EA, Kraus RJ, Farrell ML, Jordan VC, Mertz JE (2007) Estrogen-related receptor alpha1 transcriptional activities are regulated in part via the ErbB2/HER2 signaling pathway. Mol Cancer Res 5(1):71–85. https://doi.org/10.1158/1541-7786.MCR-06-0227

    Article  CAS  PubMed  Google Scholar 

  8. Deblois G, Hall JA, Perry MC, Laganiere J, Ghahremani M, Park M, Hallett M, Giguere V (2009) Genome-wide identification of direct target genes implicates estrogen-related receptor alpha as a determinant of breast cancer heterogeneity. Cancer Res 69(15):6149–6157. https://doi.org/10.1158/0008-5472.CAN-09-1251

    Article  CAS  PubMed  Google Scholar 

  9. Arany Z, Foo SY, Ma Y, Ruas JL, Bommi-Reddy A, Girnun G, Cooper M, Laznik D, Chinsomboon J, Rangwala SM, Baek KH, Rosenzweig A, Spiegelman BM (2008) HIF-independent regulation of VEGF and angiogenesis by the transcriptional coactivator PGC-1alpha. Nature 451(7181):1008–1012. https://doi.org/10.1038/nature06613

    Article  CAS  PubMed  Google Scholar 

  10. Deblois G (1812) Giguere V (2011) functional and physiological genomics of estrogen-related receptors (ERRs) in health and disease. Biochim Biophys Acta 8:1032–1040. https://doi.org/10.1016/j.bbadis.2010.12.009

    Article  CAS  Google Scholar 

  11. Kamei Y, Ohizumi H, Fujitani Y, Nemoto T, Tanaka T, Takahashi N, Kawada T, Miyoshi M, Ezaki O, Kakizuka A (2003) PPARgamma coactivator 1beta/ERR ligand 1 is an ERR protein ligand, whose expression induces a high-energy expenditure and antagonizes obesity. Proc Natl Acad Sci U S A 100(21):12378–12383. https://doi.org/10.1073/pnas.2135217100

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Schreiber SN, Knutti D, Brogli K, Uhlmann T, Kralli A (2003) The transcriptional coactivator PGC-1 regulates the expression and activity of the orphan nuclear receptor estrogen-related receptor alpha (ERRalpha). J Biol Chem 278(11):9013–9018. https://doi.org/10.1074/jbc.M212923200

    Article  CAS  PubMed  Google Scholar 

  13. Luo J, Sladek R, Carrier J, Bader JA, Richard D, Giguere V (2003) Reduced fat mass in mice lacking orphan nuclear receptor estrogen-related receptor alpha. Mol Cell Biol 23(22):7947–7956. https://doi.org/10.1128/mcb.23.22.7947-7956.2003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Leone TC, Lehman JJ, Finck BN, Schaeffer PJ, Wende AR, Boudina S, Courtois M, Wozniak DF, Sambandam N, Bernal-Mizrachi C, Chen Z, Holloszy JO, Medeiros DM, Schmidt RE, Saffitz JE, Abel ED, Semenkovich CF, Kelly DP (2005) PGC-1alpha deficiency causes multi-system energy metabolic derangements: muscle dysfunction, abnormal weight control and hepatic steatosis. PLoS Biol 3(4):e101. https://doi.org/10.1371/journal.pbio.0030101

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Teng CT, Beames B, Alex Merrick B, Martin N, Romeo C, Jetten AM (2014) Development of a stable cell line with an intact PGC-1alpha/ERRalpha axis for screening environmental chemicals. Biochem Biophys Res Commun 444(2):177–181. https://doi.org/10.1016/j.bbrc.2014.01.033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Teng CT, Hsieh JH, Zhao J, Huang R, **a M, Martin N, Gao X, Dixon D, Auerbach SS, Witt KL, Merrick BA (2017) Development of novel cell lines for high-throughput screening to detect estrogen-related receptor alpha modulators. SLAS Discov 22(6):720–731. https://doi.org/10.1177/2472555216689772

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Lynch C, Zhao J, Huang R, Kanaya N, Bernal L, Hsieh JH, Auerbach SS, Witt KL, Merrick BA, Chen S, Teng CT, **a M (2018) Identification of estrogen-related receptor alpha agonists in the Tox21 compound library. Endocrinology 159(2):744–753. https://doi.org/10.1210/en.2017-00658

    Article  CAS  PubMed  Google Scholar 

  18. Lynch C, Zhao J, Sakamuru S, Zhang L, Huang R, Witt KL, Merrick BA, Teng CT, **a M (2019) Identification of compounds that inhibit estrogen-related receptor alpha signaling using high-throughput screening assays. Molecules 24(5):841. https://doi.org/10.3390/molecules24050841

    Article  CAS  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported in part by the Intramural research program of the NCATS, NIH.

Author information

Authors and Affiliations

  1. National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA

    Caitlin Lynch, **ghua Zhao & Menghang **a

Authors
  1. Caitlin Lynch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. **ghua Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Menghang **a
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Menghang **a .

Editor information

Editors and Affiliations

  1. Center for Computational and Integrative Biology, Rutgers University, Camden, NJ, USA

    Hao Zhu

  2. National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA

    Menghang **a

Rights and permissions

Reprints and permissions

Copyright information

© 2022 This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Lynch, C., Zhao, J., **a, M. (2022). Cell-Based Assays to Identify ERR and ERR/PGC Modulators. In: Zhu, H., **a, M. (eds) High-Throughput Screening Assays in Toxicology. Methods in Molecular Biology, vol 2474. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2213-1_1

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2213-1_1

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2212-4

  • Online ISBN: 978-1-0716-2213-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation