Skip to main content

Detection and Functional Analysis of Tumor-Derived LXR Ligands

  • Protocol
  • First Online:
Tumor Immunology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1393))

Abstract

There is growing evidence highlighting the ability of nuclear receptors to control not only metabolism, but also inflammation and cancer progression. In particular liver X receptors (LXRs), the nuclear receptors physiologically involved in cholesterol homeostasis, have been shown to regulate innate and adaptive immune responses in many pathological conditions, including cancer.

We have recently demonstrated that LXR ligands (oxysterols) released by tumor cells may have an immunomodulatory role, affecting the immune cells involved in the antitumor immune response. Indeed, oxysterols inhibit the expression of the chemokine receptor CCR7 on dendritic cells (DC) in an LXR-dependent manner, thus impairing DC migration to secondary lymphoid organs, and therefore dampening the induction of successful antitumor responses.

We have resorted to direct (i.e., luciferase-based LXR activation assay) and indirect (i.e., activation of LXR target genes in dendritic cells) methods in order to assess the presence of LXR ligands (oxysterols) in tumor-conditioned media.

These two methods are also suitable to study strategies to block oxysterol release by tumor cells.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 139.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Yang X, Lamia KA, Evans RM (2007) Nuclear receptors, metabolism, and the circadian clock. Cold Spring Harb Symp Quant Biol 72:387–394

    Article  CAS  PubMed  Google Scholar 

  2. Kalaany NY, Mangelsdorf DJ (2006) LXRS and FXR: the yin and yang of cholesterol and fat metabolism. Annu Rev Physiol 68:159–191

    Article  CAS  PubMed  Google Scholar 

  3. Traversari C, Russo V (2012) Control of the immune system by oxysterols and cancer development. Curr Opin Pharmacol 12:729–735

    Article  CAS  PubMed  Google Scholar 

  4. Peet DJ, Janowski BA, Mangelsdorf DJ (1998) The LXRs: a new class of oxysterol receptors. Curr Opin Genet Dev 8:571–575

    Article  CAS  PubMed  Google Scholar 

  5. Bjorkhem I (2002) Do oxysterols control cholesterol homeostasis? J Clin Invest 110:725–730

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Murphy RC, Johnson KM (2008) Cholesterol, reactive oxygen species, and the formation of biologically active mediators. J Biol Chem 283:15521–15525

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Mast N, Annalora AJ, Lodowski DT et al (2011) Structural basis for three-step sequential catalysis by the cholesterol side chain cleavage enzyme CYP11A1. J Biol Chem 286:5607–5613

    Article  CAS  PubMed  Google Scholar 

  8. Glass CK, Rosenfeld MG (2000) The coregulator exchange in transcriptional functions of nuclear receptors. Genes Dev 14:121–141

    CAS  PubMed  Google Scholar 

  9. Jakobsson T, Treuter E, Gustafsson JA et al (2012) Liver X receptor biology and pharmacology: new pathways, challenges and opportunities. Trends Pharmacol Sci 33:394–404

    Article  CAS  PubMed  Google Scholar 

  10. Bensinger SJ, Tontonoz P (2008) Integration of metabolism and inflammation by lipid-activated nuclear receptors. Nature 454:470–477

    Article  CAS  PubMed  Google Scholar 

  11. Joseph SB, Bradley MN, Castrillo A et al (2004) LXR-dependent gene expression is important for macrophage survival and the innate immune response. Cell 119:299–309

    Article  CAS  PubMed  Google Scholar 

  12. Valledor AF, Hsu LC, Ogawa S et al (2004) Activation of liver X receptors and retinoid X receptors prevents bacterial-induced macrophage apoptosis. Proc Natl Acad Sci U S A 101:17813–17818

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. A-Gonzalez N, Bensinger SJ, Hong C et al (2009) Apoptotic cells promote their own clearance and immune tolerance through activation of the nuclear receptor LXR. Immunity 31:245–258

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Bensinger SJ, Bradley MN, Joseph SB et al (2008) LXR signaling couples sterol metabolism to proliferation in the acquired immune response. Cell 134:97–111

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Walcher D, Kummel A, Kehrle B et al (2006) LXR activation reduces proinflammatory cytokine expression in human CD4-positive lymphocytes. Arterioscler Thromb Vasc Biol 26:1022–1028

    Article  CAS  PubMed  Google Scholar 

  16. Cui G, Qin X, Wu L et al (2011) Liver X receptor (LXR) mediates negative regulation of mouse and human Th17 differentiation. J Clin Invest 121:658–670

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Villablanca EJ, Raccosta L, Zhou D et al (2010) Tumor-mediated liver X receptor-alpha activation inhibits CC chemokine receptor-7 expression on dendritic cells and dampens antitumor responses. Nat Med 16:98–105

    Article  CAS  PubMed  Google Scholar 

  18. Hanley TM, Blay Puryear W, Gummuluru S et al (2010) PPARgamma and LXR signaling inhibit dendritic cell-mediated HIV-1 capture and trans-infection. PLoS Pathog 6:e1000981

    Article  PubMed  PubMed Central  Google Scholar 

  19. Feig JE, Pineda-Torra I, Sanson M et al (2010) LXR promotes the maximal egress of monocyte-derived cells from mouse aortic plaques during atherosclerosis regression. J Clin Invest 120:4415–4424

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Villablanca EJ, Zhou D, Valentinis B et al (2008) Selected natural and synthetic retinoids impair CCR7- and CXCR4-dependent cell migration in vitro and in vivo. J Leukoc Biol 84:871–879

    Article  CAS  PubMed  Google Scholar 

  21. Bergstrom JD, Kurtz MM, Rew DJ et al (1993) Zaragozic acids: a family of fungal metabolites that are picomolar competitive inhibitors of squalene synthase. Proc Natl Acad Sci U S A 90:80–84

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Fuda H, Javitt NB, Mitamura K et al (2007) Oxysterols are substrates for cholesterol sulfotransferase. J Lipid Res 48:1343–1352

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Vincenzo Russo .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Fontana, R., Paniccia, A., Russo, V. (2016). Detection and Functional Analysis of Tumor-Derived LXR Ligands. In: Bondanza, A., Casucci, M. (eds) Tumor Immunology. Methods in Molecular Biology, vol 1393. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3338-9_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3338-9_5

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3336-5

  • Online ISBN: 978-1-4939-3338-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics