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Bioluminescence imaging of invasive intracranial xenografts: implications for translational research and targeted therapeutics of brain tumors

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Abstract

Despite decades of study, the etiology of brain cancer remains elusive. However, extensive molecular characterization of primary brain tumors has been accomplished, outlining recurrent features that are proving useful for devising targeted therapies. There are far too few patients available for comparing the efficacy of therapeutic combinations, especially when variations in dosing, frequency, and sequencing are taken into account. Consequently, there is a substantial need for increasing preclinical testing throughput using clinically relevant models. We review luminescent optical imaging for its potential in facilitating in vivo assessment of intracranial tumor growth and response to therapy in rodent orthotopic xenograft models of primary brain malignancies. We review the rationale behind the need of an in vivo model, why orthotopic tumor models displaying an invasive phenotype may be a superior choice when compared to flank-implanted tumors, and what advantages may be drawn from the use of modified cells, suitable for sequential monitoring by in vivo optical imaging. Studies show that luminescent signal correlates highly both with tumor burden and Kaplan–Meier survival curves of rodents bearing intracranial xenografts. We conclude that bioluminescent imaging is a highly sensitive technique for assessment of tumor burden, response to therapy, tumor recurrence, and behavior to salvage therapy, making it a superior option for longitudinal monitoring in intracranial rodent models of primary brain tumors.

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Authors and Affiliations

  1. Department of Neurosurgery, “Bagdasar-Arseni” Hospital, Sos. Berceni nr. 12, sector 4, 041915, Bucharest, Romania

    Eduard B. Dinca & Alexandru V. Ciurea

  2. Department of Neurosurgery, University of California, San Francisco, CA, USA

    Ramona V. Voicu

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  1. Eduard B. Dinca
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Correspondence to Eduard B. Dinca.

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Comments

Allan H. Friedman, Durham, USA

Bioluminescence imaging (BLI) of invasive intracranial xenografts: implications for translational research and targeted therapeutics of brain tumors

As the authors so well describe, we have made great progress at treating models of glioblastomas but sadly have made modest progress treating patients harboring glioblastomas. The problem is that the models do not accurately reproduce the disease. Cells grown out in cultures in media containing an artificial arrangement of growth factors are very different than cells that have developed strategies to survive in the human brain. Tumors implanted into laboratory animals are greatly influenced by the micro environment of the host tissue receiving the implant. I disagree with the author’s dismissal of the modified immune system in athymic mice. It is possible that the intrinsic immune system can become important once the tumor has been wounded by a therapeutic intervention. It is also possible that the strategies tumors use to thwart adaptive immunity are not relevant in the nascent immune system of athymic mice. Whatever the reason, it is much easier to eradicate a GBM in an athymic mouse than in a patient.

The authors describe their work and review the literature using bioluminescence to monitor the growth of intracranially implanted modified malignant xenografts. I myself have no experience with this technique, but I found the authors’ review to be quite compelling. The luciferase-transfected cells seemed to behave in a fashion similar to nontransfected cells. The transfection seemed to be stable over time in cell culture and in implanted grafts. The measured fluorescence correlated well with tumor volume as measured by in vivo MRI and histologically reconstructed tumors. The authors are correct in noting that animal MRI imaging’s equipment is expensive and somewhat cumbersome to use.

The author’s bibliography provides the interested reader with references to the transfection technique and the mechanics of measuring bioluminescence from intracranially implanted tumors. We should not lose track of the author’s point that the models of brain tumors presently available only partially replicate the human condition.

Matthias Kirsch, Dresden, Germany

The authors present a timely review of an established technology using luciferase-induced luminescence for in vivo imaging of tumor growth that still has to find its way into routine monitoring in experimental neuro-oncology.

Bioluminescence imaging is an important adjunct to the experimental armamentarium in neuro-oncology since it offers the possibility for frequent intravital measurements and quantification of tumor growth. It cannot offer morphological resolution as MRI does but is cheaper and much easier to use. BLI is limited to integral two-dimensional imaging, yet—at least in subcutaneous tumor models—bioluminescent data permitted earlier detection of tumor growth than standard caliper measurements.

Currently, it is limited due to the limited penetration depth of light and, therefore, to its principle applications in subcutaneous or intraperitoneal tumor models. However, as the current review demonstrates, experiments in small rodents with thin skulls have shown its applicability for orthotopic brain tumor investigations. Bioluminescence imaging has not reached its limits: Three-dimensional measurements using dual projections and advanced software algorithms will further improve its usability as a brain tumor monitoring device.

Ghazaleh Tabatabai, Zurich, Switzerland

The prognosis for patients with malignant gliomas has remained poor. Novel therapeutic approaches are urgently required. Before evaluating new therapeutic strategies in clinical trials, however, it is important to thoroughly examine their efficacy and toxicity in preclinical models. Today, most preclinical studies evaluating anti-glioma therapies are performed in cell culture and rodent models. Dinca and colleagues provide an overview on currently used in vitro and in vivo techniques for studying brain tumor biology. Cell culture studies either using murine or human glioma cell lines are very helpful for performing “high throughput” studies, i.e., testing different compounds or different combinations thereof in multiple parallel experiments. The main aspect of tumor biology, however, the impact of the tumor host interaction, cannot be adequately simulated in cell culture. Thus, the translational relevance of in vitro studies alone remains questionable. Clinical relevance is supported by studying the efficacy of anti-glioma therapies in mouse models. For studying orthotopic human experimental gliomas, the human glioma cells are implanted into the brains of nude mice. Migration and invasion of human glioma cells can be reliably studied in nude mice. The important aspects of immunological interactions, however, are not reflected by studies in nude mice. Therefore, some authors suggested the use of genetically modified mice by silencing a tumor suppressor gene or overexpressing an oncogene. These approaches, on the other hand, do not mimic the sporadic occurrence of malignant gliomas. Another helpful tools are syngeneic mouse models, i.e. implanting murine glioma cells into the brain of syngeneic mice, e.g., GL-261 and C57Bl/6. Taken together, even though the different mouse models have their respective limitations, it will be important for scientists to design their studies by combining different mouse models, implanting different glioma cell lines or patient-derived cells, and thus combining the strengths and reducing the limitations. Still, for assessing anti-glioma therapies in orthotopic glioma models, it will be important to use a noninvasive imaging tool for longitudinal monitoring of efficacy. In this regard, BLI and magnetic resonance imaging (MRI) are very useful. Both techniques allow close follow-up monitoring of intracranial tumor growth during therapy. Based on these studies, future preclinical assessments of anti-glioma therapies will more closely resemble clinical follow-up neuroimaging approaches in individual patients. This will hopefully further optimize preclinical assessments and enable better clinical translations.

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Dinca, E.B., Voicu, R.V. & Ciurea, A.V. Bioluminescence imaging of invasive intracranial xenografts: implications for translational research and targeted therapeutics of brain tumors. Neurosurg Rev 33, 385–394 (2010). https://doi.org/10.1007/s10143-010-0275-4

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