SpringerLink
Log in

Development of a hypoxia-activated red-emission fluorescent probe for in vivo tumor microenvironment imaging and anti-tumor therapy

  • Original Paper
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract

Hypoxia, a significant feature of the tumor microenvironment, is closely associated with tumor growth, metastasis, and drug resistance. In the field of tumor microenvironment analysis, accurately imaging and quantifying hypoxia - a critical factor associated with tumor progression, metastasis, and resistance to therapy - remains a significant challenge. Herein, a hypoxia-activated red-emission fluorescent probe, ODP, for in vivo imaging of hypoxia in the tumor microenvironment is presented. Among various imaging methods, optical imaging is particularly convenient due to its rapid response and high sensitivity. The ODP probe specifically targets nitroreductase (AzoR), an enzyme highly expressed in hypoxic cells, playing a vital role by catalyzing the cleavage of azo bonds. The optical properties of ODP exhibited excellent performance in terms of fluorescence enhancement, fluorescence lifetime (0.81 ns), and detection limit (0.86 µM) in response to SDT. Cell imaging experiments showed that ODP could effectively detect and image intracellular hypoxia and the imaging capability of ODP was studied under various conditions including cell migration, antioxidant treatment, and different incubation times. Through comprehensive in vitro and in vivo experiments, including cellular imaging and mouse tumor models, this work demonstrates the efficacy of ODP in accurately detecting and imaging hypoxia. Moreover, ODP’s potential in inducing apoptosis in cancer cells offers a promising avenue for integrating diagnostic and therapeutic strategies in cancer treatment. This innovative approach not only contributes to the understanding and assessment of tumor hypoxia but also opens new possibilities for targeted cancer therapy.

Graphical Abstract

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

Access this article

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

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Sch. 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Data availability

Not applicable.

References

  1. Lee P, Chandel NS, Simon MC (2020) Cellular adaptation to hypoxia through hypoxia inducible factors and beyond. Nat Rev Mol Cell Biol

  2. Pouyssegur J, Dayan F, Mazure N (2006) Hypoxia signalling in cancer and approaches to enforce tumour regression. Nature 441(7092):437

    Article  CAS  PubMed  Google Scholar 

  3. Semenza L, Gregg (2003) Targeting HIF-1 for cancer therapy. Nat Rev Cancer 3(10):721–732

    Article  CAS  PubMed  Google Scholar 

  4. Semenza GL (2000) HIF-1: mediator of physiological and pathophysiological responses to hypoxia. J appl Physiol 88(4):1474–1480

    Article  CAS  PubMed  Google Scholar 

  5. Hillner B, Siegel BA, Shields AF, Liu D, Gar Ee IF, Hanna NL, Stine SH, Coleman RE (2009) The impact of positron emission tomography (PET) on expected management during cancer treatment. Findings of the National Oncologic PET Registry, Cancer

    Book  Google Scholar 

  6. Martin DR, Danrad R, Herrmann K, Semelka RC, Hussain SM (2005) Magnetic resonance imaging of the gastrointestinal tract. Top Magn Reson Imaging Tmri 16(1):77–98

    Article  PubMed  Google Scholar 

  7. Shao Q, **ng B (2010) Photoactive molecules for applications in molecular imaging and cell biology. ChemInform 41(8):2835–2846

    Google Scholar 

  8. Zhao M Benhao, Yifan, Haisheng, **nyan, Zhang, Hongxin, Chaoran, Feng, A Tumor-Microenvironment-Responsive Lanthanide-Cyanine FRET Sensor for NIR-II; Luminescence-Lifetime In Situ Imaging of Hepatocellular Carcinoma

  9. Zhang F, Chen Y, Pei P, Lei Z, Zhang X, Yin D (2021) A Promising NIR-II Fluorescent Sensor for Peptide‐Mediated Long‐term Monitoring of Kidney Dysfunction, Angewandte Chemie

  10. Rauth AM, Goldberg Z, Misra V (1997) DT-Diaphorase: Possible Roles in Cancer Chemotherapy and Carcinogenesis, Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics

  11. Sandhya S Biodegradation of Azo Dyes Under Anaerobic Condition: Role of Azoreductase, Biodegradation of Azo Dyes2010

  12. Li X, Liang X, Yin J, Lin W Organic fluorescent probes for monitoring autophagy in living cells, Chemical Society Reviews

  13. Jiao Y, Zhang L, Gao X, Si W, Duan C (2020) A cofactor-substrate‐based Supramolecular fluorescent probe for the Ultrafast Detection of Nitroreductase under hypoxic conditions. Angew Chem 132(15)

  14. Thiel Z, Rivera-Fuentes P (2019) Single‐Molecule Imaging of Active Mitochondrial Nitroreductases Using a Photo‐Crosslinking Fluorescent Sensor, Angewandte Chemie International Edition

  15. Zhang S, Chen H, Wang L, Qin X, Jiang BP, Ji SC, Shen XC, Liang H (2021) A General Approach to Design Dual Ratiometric fluorescent and photoacoustic probe for quantitatively visualizing tumor hypoxia levels in vivo. Angew Chem Int Ed Engl

  16. Son S, Won M, Green O, Hananya N, Sharma A, Jeon Y, Kwak JH, Sessler JL, Shabat D, Kim JS (2019) Chemiluminescent probe for the InVitro and InVivo Imaging of Cancers Over-expressing NQO1. Angew Chem Int Ed

  17. Kwon N, Cho MK, Park SJ, Kim D, Nam SJ, Cui L, Kim HM, Yoon J (2017) An efficient two-photon fluorescent probe for human NAD(P)H:quinone oxidoreductase (hNQO1) detection and imaging in tumor cells. Chem Commun 53(3):525–528

    Article  CAS  Google Scholar 

  18. Yang YP, Qi FJ, Qian YP, Bao XZ, Zhang HC, Ma B, Dai F, Zhang SX, Zhou B (2021) Develo** Push-Pull Hydroxylphenylpolyenylpyridinium Chromophores as Ratiometric two-photon fluorescent probes for Cellular and Intravital Imaging of Mitochondrial NQO1. Anal Chem 93(4):2385–2393

    Article  CAS  PubMed  Google Scholar 

  19. Rouillere A, Johnson, Amanda E, st B, Quinn A (2017) Prasai, Bijeta, efficacious fluorescence turn-on probe for high-contrast imaging of human cells overexpressing quinone reductase activity. Chem Commun

  20. Zhang Y, Zhao W, Chen Y, Yuan H, Fang H, Yao S, Zhang C, Xu H, Li N, Liu Z, Guo Z, Zhao Q, Liang Y, He W (2021) Rational construction of a reversible arylazo-based NIR probe for cycling hypoxia imaging in vivo. Nat Commun 12(1):2772

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Wang C, Zhang S, Huang J, Cui L, Hu J, Tan S (2019) Novel designed azo substituted semi-cyanine fluorescent probe for cytochrome P450 reductase detection and hypoxia imaging in cancer cells. RSC Adv 9(37):21572–21577

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. **nwei T, Zhao, Li Y, Pan S, Wang H (2018) Ma, Near-Infrared Fluorescent Probes for Hypoxia Detection via Joint Regulated Enzymes: Design, Synthesis, and Application in Living Cells and Mice, Analytical Chemistry

  23. **ang MH, Huang H, Liu XJ, Tong ZX, Zhang CX, Wang F, Yu RQ, Jiang JH (2019) Mitochondrion-Targeting Fluorescence Probe via Reduction Induced Charge Transfer for Fast Methionine Sulfoxide Reductases Imaging, Analytical Chemistry

  24. Xu C, Zou H, Zhao Z, Zheng Z, Kwok RTK, Lam JWY, Sung HHY, Williams ID, Chen S, Zheng L (2021) B.Z. Tang, turning on Light Emission of a dark pro-aggregation‐Induced Emission Luminogen in Aqueous Media through Reductase‐Modulated Derotation. Adv NanoBiomed Res 1(4)

  25. Yuan J, Peng R, Su D, Zhang X, Yuan L (2021) Cell membranes targeted unimolecular prodrug for programmatic photodynamic-chemo therapy. Theranostics 11(7):3502–3511

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Zielonka J, Podsiad?Y R, Czerwińska M, Sikora A, Soko?Owska J, Marcinek A (2004) Color changes accompanying one-electron reduction and oxidation of the azo dyes. J Photochem Photobiology Chem 163(3):373–379

    Article  CAS  Google Scholar 

  27. Guo Z, Park S, Yoon J, Shin I (2014) Recent progress in the development of near-infrared fluorescent probes for bioimaging applications. Chem Soc Rev 43(1):16–29

    Article  PubMed  Google Scholar 

  28. Beija M, Af Onso C, Martinho J (2009) Synthesis and applications of rhodamine derivatives as fluorescent probes. Chem Soc Rev

  29. Lin Q, Bao C, Cheng S, Yang Y, Zhu L (2012) Target-activated coumarin phototriggers specifically switch on fluorescence and photocleavage upon Bonding to Thiol-Bearing protein. J Am Chem Soc 134(11):5052–5055

    Article  CAS  PubMed  Google Scholar 

  30. Wang S, Ren WX, Hou JT, Won M, Kim JS (2021) Fluorescence imaging of pathophysiological microenvironments. Chem Soc Rev 50(4)

  31. Owens EA, Henary M, Fakhri GE, Choi HS (2016) Tissue-Specific Near-Infrared Fluorescence Imaging, Accounts of Chemical Research 1731

  32. Tian Y, Li Y, Jiang W-L, Zhou D-Y, Fei J, Li C-Y (2019) In-situ imaging of azoreductase activity in the acute and chronic ulcerative colitis mice by a near-infrared fluorescent probe. Anal Chem 91(16):10901–10907

    Article  CAS  PubMed  Google Scholar 

  33. Piao W, Tsuda S, Tanaka Y, Maeda S, Liu F, Takahashi S, Kushida Y, Komatsu T, Ueno T, Terai T, Nakazawa T, Uchiyama M, Morokuma K, Nagano T, Hanaoka K (2013) Development of azo-based fluorescent probes to detect different levels of hypoxia. Angew Chem Int Ed Engl 52(49):13028–13032

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This research was supported by grants of Postdoctoral Science Foundation of China (No. 2019M651781 and 2019TQ0142) and the Innovation Fund for Technology of Nan**g University (2021).

Author information

Author notes

  1. Chen ** and Pengfei Wu contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nan**g University, No.163 **anlin Road, Nan**g, 210023, China

    Chen **, Hai-Liang Zhu & Zhen Li

  2. Pancreas Center, The First Affiliated Hospital of Nan**g Medical University, Jiangsu province hospital, Nan**g, 210029, China

    Pengfei Wu & Min Tu

Authors
  1. Chen **
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pengfei Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Min Tu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hai-Liang Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhen Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Hai-Liang Zhu or Zhen Li.

Ethics declarations

Ethical approval

All animal care and experimental protocols complied with the Animal Management Rules of the Ministry of Health of the People’s Republic of China and were approved by the Model Animal Research Center (MARC) of Nan**g University.

Conflict of interest

There is no conflict of interest to declare.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Material 1

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

**, C., Wu, P., Tu, M. et al. Development of a hypoxia-activated red-emission fluorescent probe for in vivo tumor microenvironment imaging and anti-tumor therapy. Microchim Acta 191, 217 (2024). https://doi.org/10.1007/s00604-024-06291-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00604-024-06291-7

Keywords

Search

Navigation