SpringerLink
Log in

Map** the Fate of Hypoxic Cells Using an Irreversible Fluorescent Switch

  • Protocol
  • First Online:
Hypoxia

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2755))

  • 345 Accesses

Abstract

Hypoxia has been reported to promote tumor progression and metastasis in murine models, and patients with hypoxic tumors have a worse prognosis. Besides its effect on cancer, normal processes like embryogenesis, or other pathologies such as ischemia, depend on hypoxia-regulated mechanisms. Given the degradable nature of HIF-1/2α in the presence of oxygen, defining the role of hypoxia in modeling biological processes becomes challenging when a cell enters oxygen-rich regions within a tissue. Here, we describe a unique approach to permanently mark cells that experience hypoxia with a fluorescent protein switch that is maintained even after a cell is reoxygenated. This method consists of a dual-viral delivery system that can be transduced into any mammalian cell line.

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
EUR 44.95
Price includes VAT (Germany)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
EUR 149.99
Price includes VAT (Germany)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
EUR 192.59
Price includes VAT (Germany)
  • 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. Le A, Stine ZE, Nguyen C et al (2014) Tumorigenicity of hypoxic respiring cancer cells revealed by a hypoxia-cell cycle dual reporter. Proc Natl Acad Sci 111:12486–12491

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Kim H, Lin Q, Glazer PM, Yun Z (2018) The hypoxic tumor microenvironment in vivo selects the cancer stem cell fate of breast cancer cells. Breast Cancer Res 20:1–15

    Article  Google Scholar 

  3. Fluegen G, Avivar-Valderas A, Wang Y et al (2017) Phenotypic heterogeneity of disseminated tumour cells is preset by primary tumour hypoxic microenvironments. Nat Cell Biol 19:120–132

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Kimura W, ** identifies cycling cardiomyocytes in the adult heart. Nature 523:226–230

    Article  CAS  PubMed  Google Scholar 

  5. Salceda S, Caro J (1997) Hypoxia-inducible factor 1α (HIF-1α) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. J Biol Chem 272:22642–22647

    Article  CAS  PubMed  Google Scholar 

  6. Godet I, Shin YJ, Ju JA et al (2019) Fate-map** post-hypoxic tumor cells reveals a ROS-resistant phenotype that promotes metastasis. Nat Commun 10:4862

    Article  PubMed  PubMed Central  Google Scholar 

  7. Glunde K, Shah T, Winnard PT et al (2008) Hypoxia regulates choline kinase expression through hypoxia-inducible factor-1 alpha signaling in a human prostate cancer model. Cancer Res 68:172–180

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Vasilev DS, Dubrovskaya NM, Tumanova NL, Zhuravin IA (2016) Prenatal hypoxia in different periods of embryogenesis differentially affects cell migration, neuronal plasticity, and rat behavior in postnatal ontogenesis. Front Neurosci 10:126

    Article  PubMed  PubMed Central  Google Scholar 

  9. Godet I, Doctorman S, Wu F, Gilkes DM (2022) Detection of hypoxia in cancer models: significance, challenges, and advances. Cells 11:686

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Chae Ye I, Fertig EJ, DiGiacomo JW et al (2018) Molecular portrait of hypoxia in breast cancer: a prognostic signature and novel HIF-regulated genes. Mol Cancer Res 16:1889–1901

    Article  PubMed  PubMed Central  Google Scholar 

  11. Sullivan R, Graham CH (2007) Hypoxia-driven selection of the metastatic phenotype. Cancer Metastasis Rev 26:319–331

    Article  CAS  PubMed  Google Scholar 

  12. Walsh JC, Lebedev A, Aten E et al (2014) The clinical importance of assessing tumor hypoxia: relationship of tumor hypoxia to prognosis and therapeutic opportunities. Antioxid Redox Signal 21:878–880

    Article  Google Scholar 

  13. Rocha HL, Godet I, Kurtoglu F et al (2021) A persistent invasive phenotype in post-hypoxic tumor cells is revealed by fate map** and computational modeling. iScience 24:102935

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Godet I, Mamo M, Thurnheer A et al (2021) Post-hypoxic cells promote metastatic recurrence after chemotherapy treatment in TNBC. Cancers (Basel) 13:5509

    Article  CAS  PubMed  Google Scholar 

  15. Ravi M, Paramesh V, Kaviya SR et al (2015) 3D cell culture systems: advantages and applications. J Cell Physiol 230:16–26

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Alyssa G. Weinstein & Daniele M. Gilkes

  2. Biochemistry and Molecular Biology Program, Johns Hopkins University School of Public Health, Baltimore, MD, USA

    Alyssa G. Weinstein

  3. Johns Hopkins Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA

    Daniele M. Gilkes

  4. Cancer Biology and Genetics Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY, USA

    Inês Godet

Authors
  1. Alyssa G. Weinstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniele M. Gilkes
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Inês Godet
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Inês Godet .

Editor information

Editors and Affiliations

  1. Department of Oncology Johns Hopkins, University School of Medicine, Baltimore, MD, USA

    Daniele M. Gilkes

Rights and permissions

Reprints and permissions

Copyright information

© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Weinstein, A.G., Gilkes, D.M., Godet, I. (2024). Map** the Fate of Hypoxic Cells Using an Irreversible Fluorescent Switch. In: Gilkes, D.M. (eds) Hypoxia. Methods in Molecular Biology, vol 2755. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3633-6_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3633-6_3

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3632-9

  • Online ISBN: 978-1-0716-3633-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation