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Bioluminescence imaging of hollow fibers in living animals: its application in monitoring molecular pathways

Abstract

We have applied noninvasive optical imaging technology to the in vivo hollow fiber assay, using tumor cell lines in which optical reporters are expressed in response to activation/inhibition of a specific molecular pathway. In vivo noninvasive imaging of molecular pathways in cells within hollow fibers enables a rapid and accurate evaluation of drug targets and provides useful insights to guide novel drug discovery. In this protocol we show, as an example, that a luciferase reporter, driven by the responsive element of nuclear factor NF-κB, was induced in cells within hollow fibers implanted in living mice, and a detailed procedure for in vivo bioluminescence imaging of hollow fibers is described. This approach can, in principle, be applied to image any molecular pathways of interest when appropriate reporter cells are generated. Hollow fiber encapsulation and implantation takes 2 d, and in vivo validation of reporter takes 1–2 weeks.

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Figure 1: Hollow fibers and tools for encapsulation of cells into hollow fibers and implantation of hollow fibers into mice.
Figure 2: Picture of Xenogen IVIS200 imaging system.
Figure 3: Detection of MAT B III-NF-κB-Luc cells and NF-κB reporter induction by lipopolysaccharide (LPS) in vitro.
Figure 4: Induction of NF-κB reporter by tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS) in hollow fiber in vivo.

References

  1. 1

    Hanahan, D. & Weinberg, R.A. The hallmarks of cancer. Cell 100, 57–70 (2000).

    CAS  Google Scholar 

  2. 2

    Blasberg, R.G. Molecular imaging and cancer. Mol. Cancer Ther. 2, 335–343 (2003).

    CAS  PubMed  Google Scholar 

  3. 3

    Contag, C.H., Jenkins, D., Contag, P.R. & Negrin, R.S. Use of reporter genes for optical measurements of neoplastic disease in vivo. Neoplasia 2, 41–52 (2000).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4

    Wang, W. & El-Deiry, W.S. Bioluminescent molecular imaging of endogenous and exogenous p53-mediated transcription in vitro and in vivo using an HCT116 human colon carcinoma xenograft model. Cancer Biol. Ther. 2, 196–202 (2003).

    Article  PubMed  Google Scholar 

  5. 5

    Uhrbom, L., Nerio, E. & Holland, E.C. Dissecting tumor maintenance requirements using bioluminescence imaging of cell proliferation in a mouse glioma model. Nat. Med. 10, 1257–1260 (2004).

    CAS  Article  PubMed  Google Scholar 

  6. 6

    Karin, M. & Greten, F.R. NF-κB: linking inflammation and immunity to cancer development and progression. Nat. Rev. Immunol. 5, 749–759 (2005).

    CAS  Article  Google Scholar 

  7. 7

    Zhang, G.J. et al. Bioluminescent imaging of Cdk2 inhibition in vivo. Nat. Med. 10, 643–648 (2004).

    CAS  Article  PubMed  Google Scholar 

  8. 8

    Safran, M. et al. Mouse reporter strain for noninvasive bioluminescent imaging of cells that have undergone Cre-mediated recombination. Mol. Imaging 2, 297–302 (2003).

    CAS  Article  PubMed  Google Scholar 

  9. 9

    Suggitt, M. & Bibby, M.C. 50 Years of preclinical anticancer drug screening: empirical to target-driven approaches. Clin. Cancer Res. 11, 971–981 (2005).

    CAS  Google Scholar 

  10. 10

    Bednar, B., Zhang, G.J., Williams, D.L., Hargreaves, R. & Sur, C. Optical molecular imaging in drug discovery and clinical development. Expert Opin. Drug Discov. 2, 65–85 (2007).

    CAS  Article  PubMed  Google Scholar 

  11. 11

    Hollingshead, M.G. et al. In vivo cultivation of tumor cells in hollow fibers. Life Sci. 57, 131–141 (1995).

    CAS  Article  PubMed  Google Scholar 

  12. 12

    Hall, L.A. et al. The hollow fiber assay: continued characterization with novel approaches. Anticancer Res. 20, 903–911 (2000).

    CAS  PubMed  Google Scholar 

  13. 13

    Zhang, G.J. et al. Optical imaging of tumor cells in hollow fibers: evaluation of the antitumor activities of anticancer drugs and target validation. Neoplasia 9, 652–661 (2007).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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Correspondence to Guo-Jun Zhang.

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Zhang, GJ., Chen, TB., Hargreaves, R. et al. Bioluminescence imaging of hollow fibers in living animals: its application in monitoring molecular pathways. Nat Protoc 3, 891–899 (2008). https://doi.org/10.1038/nprot.2008.52

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