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Detection of Ferroptosis by BODIPY™ 581/591 C11

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Immune Mediators in Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2108))

Abstract

Ferroptosis is a distinctive form of regulated cell death that is driven by lethal accumulation of lipid peroxides in plasma membranes. Failure to control ferroptosis has been implicated in multiple pathological conditions including cancer development, neurodegeneration, renal injury, ischemia/reperfusion injury, and T-cell immunity. Here we describe a method to detect ferroptosis by determining the amount of lipid peroxides in cellular membranes using BODIPY-C11 probe and flow cytometry. Putative role of ferroptosis in immune modulatory cells can be determined using the same method.

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References

  1. Stockwell BR, Friedmann Angeli JP, Bayir H, Bush AI, Conrad M, Dixon SJ, Fulda S, Gascon S, Hatzios SK, Kagan VE, Noel K, Jiang X, Linkermann A, Murphy ME, Overholtzer M, Oyagi A, Pagnussat GC, Park J, Ran Q, Rosenfeld CS, Salnikow K, Tang D, Torti FM, Torti SV, Toyokuni S, Woerpel KA, Zhang DD (2017) Ferroptosis: a regulated cell death nexus linking metabolism, redox biology, and disease. Cell 171(2):273–285. https://doi.org/10.1016/j.cell.2017.09.021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM, Gleason CE, Patel DN, Bauer AJ, Cantley AM, Yang WS, Morrison B 3rd, Stockwell BR (2012) Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell 149(5):1060–1072. https://doi.org/10.1016/j.cell.2012.03.042

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Yang WS, Stockwell BR (2016) Ferroptosis: death by lipid peroxidation. Trends Cell Biol 26(3):165–176. https://doi.org/10.1016/j.tcb.2015.10.014

    Article  CAS  PubMed  Google Scholar 

  4. Seong SY, Matzinger P (2004) Hydrophobicity: an ancient damage-associated molecular pattern that initiates innate immune responses. Nat Rev Immunol 4(6):469–478. https://doi.org/10.1038/nri1372

    Article  CAS  PubMed  Google Scholar 

  5. Krysko DV, Agostinis P, Krysko O, Garg AD, Bachert C, Lambrecht BN, Vandenabeele P (2011) Emerging role of damage-associated molecular patterns derived from mitochondria in inflammation. Trends Immunol 32(4):157–164. https://doi.org/10.1016/j.it.2011.01.005

    Article  CAS  PubMed  Google Scholar 

  6. Martin-Sanchez D, Ruiz-Andres O, Poveda J, Carrasco S, Cannata-Ortiz P, Sanchez-Nino MD, Ruiz Ortega M, Egido J, Linkermann A, Ortiz A, Sanz AB (2017) Ferroptosis, but not necroptosis, is important in nephrotoxic folic acid-induced AKI. J Am Soc Nephrol 28(1):218–229. https://doi.org/10.1681/ASN.2015121376

    Article  CAS  PubMed  Google Scholar 

  7. Matsushita M, Freigang S, Schneider C, Conrad M, Bornkamm GW, Kopf M (2015) T cell lipid peroxidation induces ferroptosis and prevents immunity to infection. J Exp Med 212(4):555–568. https://doi.org/10.1084/jem.20140857

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Yang WS, Sriramaratnam R, Welsch ME, Shimada K, Skouta R, Viswanathan VS, Cheah JH, Clemons PA, Shamji AF, Clish CB, Brown LM, Girotti AW, Cornish VW, Schreiber SL, Stockwell BR (2014) Regulation of ferroptotic cancer cell death by GPX4. Cell 156(1-2):317–331. https://doi.org/10.1016/j.cell.2013.12.010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Friedmann Angeli JP, Schneider M, Proneth B, Tyurina YY, Tyurin VA, Hammond VJ, Herbach N, Aichler M, Walch A, Eggenhofer E, Basavarajappa D, Radmark O, Kobayashi S, Seibt T, Beck H, Neff F, Esposito I, Wanke R, Forster H, Yefremova O, Heinrichmeyer M, Bornkamm GW, Geissler EK, Thomas SB, Stockwell BR, O'Donnell VB, Kagan VE, Schick JA, Conrad M (2014) Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice. Nat Cell Biol 16(12):1180–1191. https://doi.org/10.1038/ncb3064

    Article  CAS  PubMed  Google Scholar 

  10. Uchida K, Szweda LI, Chae HZ, Stadtman ER (1993) Immunochemical detection of 4-hydroxynonenal protein adducts in oxidized hepatocytes. Proc Natl Acad Sci U S A 90(18):8742–8746

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Yagi K (1998) Simple assay for the level of total lipid peroxides in serum or plasma. Methods Mol Biol 108:101–106. https://doi.org/10.1385/0-89603-472-0:101

    Article  CAS  PubMed  Google Scholar 

  12. Drummen GP, van Liebergen LC, Op den Kamp JA, Post JA (2002) C11-BODIPY(581/591), an oxidation-sensitive fluorescent lipid peroxidation probe: (micro)spectroscopic characterization and validation of methodology. Free Radic Biol Med 33(4):473–490

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by National Institute of Health Grants 1SC2NS104275 to W.S.Y.

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Authors and Affiliations

  1. Department of Biological Sciences, St. John’s University, Queens, NY, USA

    Alejandra M. Martinez, Ahryun Kim & Wan Seok Yang

Authors
  1. Alejandra M. Martinez
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  2. Ahryun Kim
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  3. Wan Seok Yang
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Corresponding author

Correspondence to Wan Seok Yang .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, St. John’s University, Queens, NY, USA

    Ivana Vancurova

  2. Department of Biological Sciences, St. John’s University, Queens, NY, USA

    Yan Zhu

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Martinez, A.M., Kim, A., Yang, W.S. (2020). Detection of Ferroptosis by BODIPY™ 581/591 C11. In: Vancurova, I., Zhu, Y. (eds) Immune Mediators in Cancer. Methods in Molecular Biology, vol 2108. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0247-8_11

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  • DOI: https://doi.org/10.1007/978-1-0716-0247-8_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0246-1

  • Online ISBN: 978-1-0716-0247-8

  • eBook Packages: Springer Protocols

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