Skip to main content
SpringerLink
Log in

5-ALA kinetics in meningiomas: analysis of tumor fluorescence and PpIX metabolism in vitro and comparative analyses with high-grade gliomas

  • Laboratory Investigation
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

Introduction

Although the utility 5-aminolevulinic acid (5-ALA)-mediated fluorescence-guided surgery (FGS) in meningiomas is increasingly discussed, data about the kinetics of protoporphyrin IX (PpIX) and tumor fluorescence are sparse.

Methods

PpIX kinetics after exposition to varying 5-ALA doses (12.5–150 µg/ml) was analyzed in two immortalized as well as primary WHO grade I and II meningioma and U87 high-grade glioma cell lines. Expression of FECH, ABCB6 and ABCG2 was investigated by quantitative real-time PCR.

Results

Fluorescence in Ben-Men 1 and primary WHO grade I/II meningioma increased with rising 5-ALA doses up to 100 µg/ml but then showed a saturation effect. However, decrease of fluorescence was slower after 150 than after 100 µg/ml 5-ALA. Fluorescence in U87 cells marginally increased with rising 5-ALA doses. Kinetics of the fluorescence in Ben-Men 1 cells did not differ from primary meningioma cells after 25–150 µg/ml 5-ALA (p > .05, each). No difference was found when comparing the fluorescence between primary grade I and II meningiomas after any 5-ALA dosage (p > .05, each). No relevant fluorescence was found in IOMM-Lee cells. Expression of FECH, ABCB6 and ABCG2 as well as PpIX export differed between all analyzed cell lines but were not connected to fluorescence.

Conclusions

Eligibility of established meningioma cell lines for in-vitro analyzes of tumor fluorescence significantly differs. Fluorescence in Ben-Men 1 and primary meningioma cell lines but less in IOMM Lee cells is 5-ALA dose-dependent, encouraging in-situ trials to encounter currently discussed shortcomings of FGS in meningiomas. Fluorescence is not related to expression of FECH, ABCB6 and ABCG2.

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

Similar content being viewed by others

Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Code availability

No custom software code was written for data analysis in this study.

References

  1. Ewelt C, Nemes A, Senner V, Wolfer J, Brokinkel B, Stummer W, Holling M (2015) Fluorescence in neurosurgery: Its diagnostic and therapeutic use. Review of the literature. J Photochem Photobiol B 148:302–309. https://doi.org/10.1016/j.jphotobiol.2015.05.002

    Article  CAS  PubMed  Google Scholar 

  2. Stummer W, Pichlmeier U, Meinel T, Wiestler OD, Zanella F, Reulen HJ, Group AL-GS (2006) Fluorescence-guided surgery with 5-aminolevulinic acid for resection of malignant glioma: a randomised controlled multicentre phase III trial. Lancet Oncol 7:392–401. https://doi.org/10.1016/S1470-2045(06)70665-9

    Article  CAS  PubMed  Google Scholar 

  3. Foster N, Eljamel S (2016) ALA-induced fluorescence image guided surgery of meningiomas: a meta-analyses. Photodiagnosis Photodyn Ther 15:73–78. https://doi.org/10.1016/j.pdpdt.2016.05.006

    Article  PubMed  Google Scholar 

  4. Valdes PA, Millesi M, Widhalm G, Roberts DW (2019) 5-aminolevulinic acid induced protoporphyrin IX (ALA-PpIX) fluorescence guidance in meningioma surgery. J Neurooncol 141:555–565. https://doi.org/10.1007/s11060-018-03079-7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Gousias K, Schramm J, Simon M (2016) The Simpson grading revisited: aggressive surgery and its place in modern meningioma management. J Neurosurg 125:551–560. https://doi.org/10.3171/2015.9.JNS15754

    Article  PubMed  Google Scholar 

  6. Spille DC, Hess K, Bormann E, Sauerland C, Brokinkel C, Warneke N, Mawrin C, Paulus W, Stummer W, Brokinkel B (2020) Risk of tumor recurrence in intracranial meningiomas: comparative analyses of the predictive value of the postoperative tumor volume and the Simpson classification. J Neurosurg. https://doi.org/10.3171/2020.4.JNS20412

    Article  PubMed  Google Scholar 

  7. Cornelius JF, Slotty PJ, El Khatib M, Giannakis A, Senger B, Steiger HJ (2014) Enhancing the effect of 5-aminolevulinic acid based photodynamic therapy in human meningioma cells. Photodiagnosis Photodyn Ther 11:1–6. https://doi.org/10.1016/j.pdpdt.2014.01.001

    Article  CAS  PubMed  Google Scholar 

  8. El-Khatib M, Tepe C, Senger B, Dibue-Adjei M, Riemenschneider MJ, Stummer W, Steiger HJ, Cornelius JF (2015) Aminolevulinic acid-mediated photodynamic therapy of human meningioma: an in vitro study on primary cell lines. Int J Mol Sci 16:9936–9948. https://doi.org/10.3390/ijms16059936

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Kaneko S, Brokinkel B, Suero Molina E, Warneke N, Holling M, Bunk EC, Hess K, Senner V, Paulus W, Stummer W (2020) Real-time in vivo kinetics of protoporphyrin IX after administration of 5-aminolevulinic acid in meningiomas and comparative analyses with glioblastomas. Acta Neurochir (Wien). https://doi.org/10.1007/s00701-020-04353-2

    Article  Google Scholar 

  10. Puttmann S, Senner V, Braune S, Hillmann B, Exeler R, Rickert CH, Paulus W (2005) Establishment of a benign meningioma cell line by hTERT-mediated immortalization. Lab Invest 85:1163–1171. https://doi.org/10.1038/labinvest.3700307

    Article  CAS  PubMed  Google Scholar 

  11. Lee WH (1990) Characterization of a newly established malignant meningioma cell line of the human brain: IOMM-Lee. Neurosurgery 27:389–395; discussion 396

    Article  CAS  PubMed  Google Scholar 

  12. Hardy SJ, Christodoulides M, Weller RO, Heckels JE (2000) Interactions of Neisseria meningitidis with cells of the human meninges. Mol Microbiol 36:817–829. https://doi.org/10.1046/j.1365-2958.2000.01923.x

    Article  CAS  PubMed  Google Scholar 

  13. Sun W, Kajimoto Y, Inoue H, Miyatake S, Ishikawa T, Kuroiwa T (2013) Gefitinib enhances the efficacy of photodynamic therapy using 5-aminolevulinic acid in malignant brain tumor cells. Photodiagnosis Photodyn Ther 10:42–50. https://doi.org/10.1016/j.pdpdt.2012.06.003

    Article  CAS  PubMed  Google Scholar 

  14. Millesi M, Kiesel B, Mischkulnig M, Martinez-Moreno M, Wohrer A, Wolfsberger S, Knosp E, Widhalm G (2016) Analysis of the surgical benefits of 5-ALA-induced fluorescence in intracranial meningiomas: experience in 204 meningiomas. J Neurosurg 125:1408–1419. https://doi.org/10.3171/2015.12.JNS151513

    Article  CAS  PubMed  Google Scholar 

  15. Potapov AA, Goryaynov SA, Danilov GV, Chelushkin DM, Okhlopkov VA, Shimanskiy VN, Beshplav ST, Poshataev VK, Shishkina LV, Zakharova NE, Spallone A, Savel’eva TA, Loshchenov VB (2018) [Intraoperative fluorescence diagnostics in surgery of intracranial meningiomas: analysis of 101 cases]. Zh Vopr Neirokhir Im N N Burdenko 82:17–29. https://doi.org/10.17116/oftalma201882217-29

    Article  CAS  PubMed  Google Scholar 

  16. Coluccia D, Fandino J, Fujioka M, Cordovi S, Muroi C, Landolt H (2010) Intraoperative 5-aminolevulinic-acid-induced fluorescence in meningiomas. Acta Neurochir (Wien) 152:1711–1719. https://doi.org/10.1007/s00701-010-0708-4

    Article  Google Scholar 

  17. Brokinkel B, Kroger S, Senner V, Jeibmann A, Karst U, Stummer W (2018) Visualizing protoporphyrin IX formation in the dura tail of meningiomas by mass spectrometry imaging. Acta Neurochir (Wien) 160:1433–1437. https://doi.org/10.1007/s00701-018-3488-x

    Article  Google Scholar 

  18. Knipps J, Beseoglu K, Kamp M, Fischer I, Felsberg J, Neumann LM, Steiger HJ, Cornelius JF (2017) Fluorescence behavior and dural infiltration of meningioma analyzed by 5-aminolevulinic acid-based fluorescence: operating microscope versus mini-spectrometer. World Neurosurg 108:118–127. https://doi.org/10.1016/j.wneu.2017.08.140

    Article  PubMed  Google Scholar 

  19. Turcotte EL, Rahme RJ, Merrill SA, Hess RA, Lettieri SC, Bendok BR (2020) Three applications of 5-aminolevulinic acid (5-ALA) for microsurgical resection of meningiomas. World Neurosurg. https://doi.org/10.1016/j.wneu.2020.03.178

    Article  PubMed  Google Scholar 

  20. Teng L, Nakada M, Zhao SG, Endo Y, Furuyama N, Nambu E, Pyko IV, Hayashi Y, Hamada JI (2011) Silencing of ferrochelatase enhances 5-aminolevulinic acid-based fluorescence and photodynamic therapy efficacy. Br J Cancer 104:798–807. https://doi.org/10.1038/bjc.2011.12

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Zhao SG, Chen XF, Wang LG, Yang G, Han DY, Teng L, Yang MC, Wang DY, Shi C, Liu YH, Zheng BJ, Shi CB, Gao X, Rainov NG (2013) Increased expression of ABCB6 enhances protoporphyrin IX accumulation and photodynamic effect in human glioma. Ann Surg Oncol 20:4379–4388. https://doi.org/10.1245/s10434-011-2201-6

    Article  PubMed  Google Scholar 

  22. Hefti M, Holenstein F, Albert I, Looser H, Luginbuehl V (2011) Susceptibility to 5-aminolevulinic acid based photodynamic therapy in WHO I meningioma cells corresponds to ferrochelatase activity. Photochem Photobiol 87:235–241. https://doi.org/10.1111/j.1751-1097.2010.00821.x

    Article  CAS  PubMed  Google Scholar 

  23. Valdes PA, Bekelis K, Harris BT, Wilson BC, Leblond F, Kim A, Simmons NE, Erkmen K, Paulsen KD, Roberts DW (2014) 5-Aminolevulinic acid-induced protoporphyrin IX fluorescence in meningioma: qualitative and quantitative measurements in vivo. Neurosurgery 10 Suppl 1:74–82; discussion 82 – 73. https://doi.org/10.1227/NEU.0000000000000117

Download references

Acknowledgements

We cordially thank Hans-Joachim Schnittler, Institute of Anatomy and Vascular Biology, University Hospital Muenster, Muenster, Germany, for kindly providing the necessary access to the microplate reader. We further thank Christian Mawrin, Institute of Neuropathology, Magdeburg, Germany, for kindly providing IOMM-Lee Meningioma cell lines.

Funding

Partial financial support was received from the Walter Schulz Stiftung, Germany.

Author information

Author notes

  1. Walter Stummer, Volker Senner and Benjamin Brokinkel have contributed equally to this work.

Authors and Affiliations

  1. Department of Neurosurgery, University Hospital Münster, Albert-Schweitzer-Campus 1, Building A1, 48149, Münster, Germany

    Eva Christine Bunk, Walter Stummer & Benjamin Brokinkel

  2. Institute of Neuropathology, University Hospital Münster, Münster, Germany

    Andrea Wagner & Volker Senner

Authors
  1. Eva Christine Bunk
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrea Wagner
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Walter Stummer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Volker Senner
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Benjamin Brokinkel
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by ECB, BB and VS. The first draft of the manuscript was written by BB and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Benjamin Brokinkel.

Ethics declarations

Conflict of interest

The authors have no financial or non-financial interests to disclose.

Ethical approval

All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. The study was approved by the local ethics committee and permitted by the patients in each single case (2019-296-f-S).

Consent to participate/for publication

Informed consent was obtained from all individual participants included in the study.

Additional information

Publisher’s Note

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

Supplementary Information

Below is the link to the electronic supplementary material.

Suppl. Figure 1: Phase contrast microscopy of primary meningioma cells.

Cells could adhere to different coatings including Matrigel (A), PLL plus mLaminin (B) and PLL plus hLaminin (C) as well as to culture plates without any coating and could be kept over several passages. Cells could also be kept in culture for several passages following kryoconservation (D) (div.=days in vitro, h = human, m = mouse, PLL = poly-L-Lysine) (A – C: WHO grade II, atypical meningioma, MIB1 Proliferation-index ND; D: WHO grade I, meningothelial meningioma, MIB1 Proliferation-index 5%). (TIF 17932 kb)

Suppl. Figure 2: Fluorescence intensity in negative controls

. As expected, no relevant fluorescence was detected in negative controls. (TIF 7922 kb)

Suppl. Figure 3: Fluorescence of primary and immortalized meningioma cell lines in comparative analyses.

Fluorescence intensity of Ben-Men 1 cells at 0, 2, 4 and 6 hours was similar as compared to primary meningioma cells after 25–150 µg/ml 5-ALA (p > .05, each). On the other hand, fluorescence intensity between primary WHO grade II meningioma cell lines and IOMM-Lee cells began to differ at 0 and 2 hours after incubation (p = .025 and 0.053) with 25 µg/ml 5-ALA. At 50 µg/ml 5-ALA, fluorescence intensity between both cell lines significantly differed at all time points (p = .036) with a similar trend at doses of 100 and 150 mg/ml 5-ALA (p = .083, each). For better overview exemplary data shown only for 50 µg/ml 5-ALA. (TIF 7523 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bunk, E.C., Wagner, A., Stummer, W. et al. 5-ALA kinetics in meningiomas: analysis of tumor fluorescence and PpIX metabolism in vitro and comparative analyses with high-grade gliomas. J Neurooncol 152, 37–46 (2021). https://doi.org/10.1007/s11060-020-03680-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11060-020-03680-9

Keywords

Search

Navigation