Skip to main content
SpringerLink
Log in

Molecular Imaging of Glucose Metabolism for Intraoperative Fluorescence Guidance During Glioma Surgery

  • Research Article
  • Published:
Molecular Imaging and Biology Aims and scope Submit manuscript

Abstract

Purpose

This study evaluated the use of molecular imaging of fluorescent glucose analog 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) as a discriminatory marker for intraoperative tumor border identification in a murine glioma model.

Procedures

2-NBDG was assessed in GL261 and U251 orthotopic tumor-bearing mice. Intraoperative fluorescence of topical and intravenous 2-NBDG in normal and tumor regions was assessed with an operating microscope, handheld confocal laser scanning endomicroscope (CLE), and benchtop confocal laser scanning microscope (LSM). Additionally, 2-NBDG fluorescence in tumors was compared with 5-aminolevulinic acid–induced protoporphyrin IX fluorescence.

Results

Intravenously administered 2-NBDG was detectable in brain tumor and absent in contralateral normal brain parenchyma on wide-field operating microscope imaging. Intraoperative and benchtop CLE showed preferential 2-NBDG accumulation in the cytoplasm of glioma cells (mean [SD] tumor-to-background ratio of 2.76 [0.43]). Topically administered 2-NBDG did not create sufficient tumor-background contrast for wide-field operating microscope imaging or under benchtop LSM (mean [SD] tumor-to-background ratio 1.42 [0.72]). However, topical 2-NBDG did create sufficient contrast to evaluate cellular tissue architecture and differentiate tumor cells from normal brain parenchyma. Protoporphyrin IX imaging resulted in a more specific delineation of gross tumor margins than intravenous or topical 2-NBDG and a significantly higher tumor-to-normal-brain fluorescence intensity ratio.

Conclusion

After intravenous administration, 2-NBDG selectively accumulated in the experimental brain tumors and provided bright contrast under wide-field fluorescence imaging with a clinical-grade operating microscope. Topical 2-NBDG was able to create a sufficient contrast to differentiate tumor from normal brain cells on the basis of visualization of cellular architecture with CLE. 5-Aminolevulinic acid demonstrated superior specificity in outlining tumor margins and significantly higher tumor background contrast. Given the nontoxicity of 2-NBDG, its use as a topical molecular marker for noninvasive in vivo intraoperative microscopy is encouraging and warrants further clinical evaluation.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.
Fig. 6.

Similar content being viewed by others

Abbreviations

2-NBDG:

2-(N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl) amino)-2-deoxyglucose

5 ALA:

5-Aminolevulinic Acid

FDG:

18F labeled 2-fluoro-2-deoxy-D-glucose

CLE:

Confocal laser endomicroscope

IV:

Intravenous

PET:

Positron emission tomography

PpIX:

Protoporphyrin IX

RFP:

Red fluorescent protein

References

  1. Carlsson SK, Brothers SP, Wahlestedt C (2014) Emerging treatment strategies for glioblastoma multiforme. EMBO Molecular Medicine 6:1359–1370

    Article  CAS  Google Scholar 

  2. Lacroix M, Abi-Said D, Fourney DR, Gokaslan ZL, Shi W, DeMonte F, Lang FF, McCutcheon IE, Hassenbusch SJ, Holland E, Hess K, Michael C, Miller D, Sawaya R (2001) A multivariate analysis of 416 patients with glioblastoma multiforme: prognosis, extent of resection, and survival. J Neurosurg 95:190–198

    Article  CAS  Google Scholar 

  3. Belykh E, Martirosyan NL, Yagmurlu K, et al. (2016) Intraoperative Fluorescence Imaging for Personalized Brain Tumor Resection: Current State and Future Directions Frontiers in Surgery 3

  4. Lakomkin N, Hadjipanayis CG (2019) The use of spectroscopy handheld tools in brain tumor surgery: current evidence and techniques. Front Surg 6:30

    Article  Google Scholar 

  5. Lakomkin N, Hadjipanayis CG (2018) Fluorescence-guided surgery for high-grade gliomas. J Surg Oncol 118:356–361

    Article  Google Scholar 

  6. Cheng Z, Levi J, Xiong Z, Gheysens O, Keren S, Chen X, Gambhir SS (2006) Near-infrared fluorescent deoxyglucose analogue for tumor optical imaging in cell culture and living mice. Bioconjug Chem 17:662–669

    Article  CAS  Google Scholar 

  7. Warburg O, Wind F, Negelein E (1927) The metabolism of tumors in the body. J Gen Physiol 8:519–530

    Article  CAS  Google Scholar 

  8. Wahl RL, Cody RL, Hutchins GD, Mudgett EE (1991) Primary and metastatic breast carcinoma: initial clinical evaluation with PET with the radiolabeled glucose analogue 2-[F-18]-fluoro-2-deoxy-D-glucose. Radiology 179:765–770

    Article  CAS  Google Scholar 

  9. Avril N, Menzel M, Dose J, Schelling M, Weber W, Jänicke F, Nathrath W, Schwaiger M (2001) Glucose metabolism of breast cancer assessed by 18F-FDG PET: histologic and immunohistochemical tissue analysis. J Nucl Med 42:9–16

    CAS  PubMed  Google Scholar 

  10. Cox BL, Mackie TR, Eliceiri KW (2015) The sweet spot: FDG and other 2-carbon glucose analogs for multi-modal metabolic imaging of tumor metabolism. Am J Nucl Med Mol Imaging 5:1–13

    PubMed  Google Scholar 

  11. O'Neil RG, Wu L, Mullani N (2005) Uptake of a fluorescent deoxyglucose analog (2-NBDG) in tumor cells. Mol Imaging Biol 7:388–392

    Article  Google Scholar 

  12. Millon SR, Ostrander JH, Brown JQ, Raheja A, Seewaldt VL, Ramanujam N (2011) Uptake of 2-NBDG as a method to monitor therapy response in breast cancer cell lines. Breast Cancer Res Treat 126:55–62

    Article  CAS  Google Scholar 

  13. Flavahan WA, Wu Q, Hitomi M, Rahim N, Kim Y, Sloan AE, Weil RJ, Nakano I, Sarkaria JN, Stringer BW, Day BW, Li M, Lathia JD, Rich JN, Hjelmeland AB (2013) Brain tumor initiating cells adapt to restricted nutrition through preferential glucose uptake. Nat Neurosci 16:1373–1382

    Article  CAS  Google Scholar 

  14. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682

    Article  CAS  Google Scholar 

  15. Yoshioka K, Takahashi H, Homma T, Saito M, Oh KB, Nemoto Y, Matsuoka H (1996) A novel fluorescent derivative of glucose applicable to the assessment of glucose uptake activity of Escherichia coli. Biochim Biophys Acta 1289:5–9

    Article  Google Scholar 

  16. Barros LF, Courjaret R, Jakoby P, Loaiza A, Lohr C, Deitmer JW (2009) Preferential transport and metabolism of glucose in Bergmann glia over Purkinje cells: a multiphoton study of cerebellar slices. Glia 57:962–970

    Article  CAS  Google Scholar 

  17. Valdes-Sosa PA, Zhou H, Luby-Phelps K, et al. (2009) Dynamic near-infrared optical imaging of 2-deoxyglucose uptake by intracranial glioma of Athymic mice. PLoS ONE 4

  18. Yamada K, Saito M, Matsuoka H, Inagaki N (2007) A real-time method of imaging glucose uptake in single, living mammalian cells. Nat Protoc 2:753–762

    Article  CAS  Google Scholar 

  19. Sheth RA, Josephson L, Mahmood U (2009) Evaluation and clinically relevant applications of a fluorescent imaging analog to fluorodeoxyglucose positron emission tomography. J Biomed Opt 14:064014

    Article  Google Scholar 

  20. La Rocca G, Sabatino G, Menna G et al (2020) 5-Aminolevulinic acid false positives in cerebral Neuro-oncology: not all that is fluorescent is tumor. A case-based update and literature review. World Neurosurgery 137:187–193

    Article  Google Scholar 

  21. Müther M, Stummer W (2019) Ependymal fluorescence in fluorescence-guided resection of malignant glioma: a systematic review. Acta Neurochir 162:365–372

    Article  Google Scholar 

  22. Hebeda KM, Saarnak AE, Olivo M, Sterenborg HJCM, Wolbers JG (1998) 5-Aminolevulinic acid induced endogenous porphyrin fluorescence in 9L and C6 brain tumours and in the normal rat brain. Acta Neurochir 140:503–513

    Article  CAS  Google Scholar 

  23. Belykh E, Miller EJ, Hu D, Martirosyan NL, Woolf EC, Scheck AC, Byvaltsev VA, Nakaji P, Nelson LY, Seibel EJ, Preul MC (2018) Scanning fiber endoscope improves detection of 5-aminolevulinic acid-induced protoporphyrin IX fluorescence at the boundary of infiltrative Glioma. World Neurosurg 113:e51–e69

    Article  Google Scholar 

  24. Folaron M, Strawbridge R, Samkoe KS, Filan C, Roberts DW, Davis SC (2018) Elucidating the kinetics of sodium fluorescein for fluorescence-guided surgery of glioma. J Neurosurg 131:724–734

    Article  Google Scholar 

  25. Belykh E, Miller EJ, Patel AA, Yazdanabadi MI, Martirosyan NL, Yağmurlu K, Bozkurt B, Byvaltsev VA, Eschbacher JM, Nakaji P, Preul MC (2018) Diagnostic accuracy of a confocal laser endomicroscope for in vivo differentiation between normal injured and tumor tissue during fluorescein-guided glioma resection: laboratory investigation. World Neurosurg 115:e337–e348

  26. Martirosyan NL, Georges J, Eschbacher JM, Cavalcanti DD, Elhadi AM, Abdelwahab MG, Scheck AC, Nakaji P, Spetzler RF, Preul MC (2014) Potential application of a handheld confocal endomicroscope imaging system using a variety of fluorophores in experimental gliomas and normal brain. Neurosurg Focus 36:E16

    Article  Google Scholar 

  27. Martirosyan NL, Eschbacher JM, Kalani MY et al (2016) Prospective evaluation of the utility of intraoperative confocal laser endomicroscopy in patients with brain neoplasms using fluorescein sodium: experience with 74 cases. Neurosurg Focus 40:E11

    Article  Google Scholar 

  28. Nitin N, Carlson AL, Muldoon T, El-Naggar AK, Gillenwater A, Richards-Kortum R (2009) Molecular imaging of glucose uptake in oral neoplasia following topical application of fluorescently labeled deoxy-glucose. Int J Cancer 124:2634–2642

    Article  CAS  Google Scholar 

  29. Thekkek N, Maru DM, Polydorides AD, Bhutani MS, Anandasabapathy S, Richards-Kortum R (2011) Pre-clinical evaluation of fluorescent deoxyglucose as a topical contrast agent for the detection of Barrett’s-associated neoplasia during confocal imaging. Technol Cancer Res Treat 10:431–441

    Article  CAS  Google Scholar 

  30. Yokoyama H, Sasaki A, Yoshizawa T, Kijima H, Hakamada K, Yamada K (2016) Imaging hamster model of bile duct cancer in vivo using fluorescent L-glucose derivatives. Hum Cell 29:111–121

  31. Pal R, Villarreal P, Qiu S, Vargas G (2018) In-vivo topical mucosal delivery of a fluorescent deoxy-glucose delineates neoplasia from normal in a preclinical model of oral epithelial neoplasia. Sci Rep 8:9760

Download references

Acknowledgments

The authors thank the staff of Neuroscience Publications at Barrow Neurological Institute for assistance with manuscript preparation. We thank Carl Zeiss, AG, Oberkochen, Germany, for providing the LSM 710 confocal laser scanning microscope.

Funding

The Barrow Neurological Foundation and the Newsome Chair in Neurosurgery Research (to MCP). The confocal laser endomicroscope was provided by Zeiss. Zeiss did not have any contribution or effect on the idea, study design, data collection, analysis, or paper preparation.

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Barrow Neurological Institute, St. Joseph’s Hospital and Medical Center, Phoenix, AZ, USA

    Evgenii Belykh, Jubran H. Jubran, Laeth L. George, Liudmila Bardonova & Mark C. Preul

  2. Department of Neuro-Oncology Research, Barrow Neurological Institute, St. Joseph’s Hospital and Medical Center, Phoenix, AZ, USA

    Deborah R. Healey, Chad C. Quarles & Adrienne C. Scheck

  3. Department of Neurosurgery, Philadelphia College of Osteopathic Medicine, Philadelphia, PA, USA

    Joseph F. Georges

  4. Department of Neuropathology, Barrow Neurological Institute, St. Joseph’s Hospital and Medical Center, Phoenix, AZ, USA

    Jennifer M. Eschbacher

  5. Ivy Brain Tumor Center, Barrow Neurological Institute, St. Joseph’s Hospital and Medical Center, Phoenix, AZ, USA

    Shwetal Mehta

  6. Department of Neurosurgery, Banner Health-University of Arizona College of Medicine, Phoenix, AZ, USA

    Peter Nakaji

Authors
  1. Evgenii Belykh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jubran H. Jubran
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Laeth L. George
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Liudmila Bardonova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Deborah R. Healey
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Joseph F. Georges
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Chad C. Quarles
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jennifer M. Eschbacher
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shwetal Mehta
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Adrienne C. Scheck
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Peter Nakaji
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Mark C. Preul
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mark C. Preul.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s Note

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

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Belykh, E., Jubran, J.H., George, L.L. et al. Molecular Imaging of Glucose Metabolism for Intraoperative Fluorescence Guidance During Glioma Surgery. Mol Imaging Biol 23, 586–596 (2021). https://doi.org/10.1007/s11307-021-01579-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11307-021-01579-z

Key words

Search

Navigation