A screening platform for glioma growth and invasion using bioluminescence imaging

Laboratory investigation

Hong Zhao M.D., Ph.D. 1 , Carol Tang Ph.D. 2 , Kemi Cui M.D., Ph.D. 1 , Beng-Ti Ang M.D. 3 , 4 and Stephen T. C. Wong Ph.D. 1
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  • 1 Department of Radiology, The Methodist Hospital and The Center for Biotechnology and Informatics, The Methodist Hospital Research Institute, Weill Medical College, Cornell University, Houston, Texas;
  • 2 Departments of Research and
  • 3 Neurosurgery, National Neuroscience Institute and Duke-National University of Singapore Graduate Medical School; and
  • 4 Singapore Institute for Clinical Sciences, Agency for Science, Technolology and Research, Singapore
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Object

The study of tumor cell growth and invasion in cancer biology is often limited by the inability to visualize tumor cell behavior in real time in animal models. The authors provide evidence that glioma cells are heterogeneous, with a subset responsible for increased invasiveness. The use of bioluminescence (BL) imaging to investigate dynamic aspects of glioma progression are discussed.

Methods

Glioblastoma multiforme–initiating cells were generated under conditions typically used to sustain neural stem cells. The invasiveness potential was determined using a Matrigel chamber. The presence of an “invasiveness gene signature” that correlated with patient survival outcome was ascertained through microarray gene expression analysis. To measure invasiveness, the authors devised a method focussed on BL imaging and tested it in vitro and in vivo using a zebrafish xenograft model. Bioluminescence imaging signals were verified using known inhibitors of glioma growth: AEE788, N-[(3,5-Difluorophenyl)acetyl]-L-alanyl-2-phenylglycine-1,1-dimethylethyl ester, and compound E.

Results

The authors' data support the idea that glioblastoma multiforme–initiating cells are heterogeneous and possess an invasive subset; BL imaging was used as a readout method to assess this invasive subset. The in vitro data obtained using a known glioma growth inhibitor, AEE788, showed that BL imaging could detect cellular movement and invasion even before overall cell death was detectable on conventional viability assays. Further work using a zebrafish tumor xenograft model supported the efficacy of BL imaging in monitoring changes in tumor load.

Conclusions

The authors used optically transparent zebrafish and high-resolution confocal imaging to track tumor growth in vivo and demonstrate the efficacy of this model for screening antitumor and antiangiogenic compounds. The integration of zebrafish transgenic technology into human cancer biological studies may aid in the development of cancer models targeting specific organs, tissues, or cell types within tumors. Zebrafish could also provide a cost-effective means for the rapid development of therapeutic agents directed at blocking tumor growth and invasion.

Abbreviations used in this paper: BL = bioluminescence; DAPT = N-[(3,5-Difluorophenyl)acetyl]-L-alanyl-2-phenylglycine-1,1-dimethylethyl ester; DMSO = dimethyl sulfoxide; IGS = invasiveness gene signature; GBM = glioblastoma multiforme.

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Contributor Notes

Address correspondence to: Stephen T. C. Wong, Ph.D., Department of Radiology, The Methodist Hospital, Weill Cornell Medical College, 6565 Fannin Street B5-022, Houston, Texas 77030. email: stwong@tmhs.org.
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