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Imaging the intracellular distribution of tyrosine kinase inhibitors in living cells with quantitative hyperspectral stimulated Raman scattering

Nature Chemistry volume 6, pages 614–622 (2014)Cite this article

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Abstract

ABL1 tyrosine-kinase inhibitors (TKI) are front-line therapy for chronic myelogenous leukaemia and are among the best-known examples of targeted cancer therapeutics. However, the dynamic uptake into cells of TKIs of low molecular weight and their intracellular behaviour is unknown because of the difficulty of observing non-fluorescent small molecules at subcellular resolution. Here we report the direct label-free visualization and quantification of two TKI drugs (imatinib and nilotinib) inside living cells using hyperspectral stimulated Raman scattering imaging. Concentrations of both drugs were enriched over 1,000-fold in lysosomes as a result of their lysosomotropic properties. In addition, low solubility appeared to contribute significantly to the surprisingly large accumulation of nilotinib. We further show that the lysosomal trapping of imatinib was reduced more than tenfold when chloroquine is used simultaneously, which suggests that chloroquine may increase the efficacy of TKIs through lysosome-mediated drug–drug interaction in addition to the commonly proposed autophagy-inhibition mechanism.

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Figure 1: Structure and spectral properties of five drug molecules: imatinib, nilotinib, chloroquine, GNF-2 and GNF-5.
Figure 2: hsSRS microscopy reveals enrichment of drugs in living cells: the SRS spectra of the bright spots in drug-treated cells match the SRS spectra of the drug in solution, but differ from that of cytosol.
Figure 3: Accumulations of drugs in lysosomes are confirmed by simultaneous two-photon fluorescence imaging of lysotracker and SRS imaging of drug accumulation.
Figure 4: hsSRS imaging of intracellular uptake of the GNF-2 and GNF-5 drugs shows that only cytoplasm spectra have moderate intensity increases at ~1,600 cm−1 caused by drug accumulation.
Figure 5: Time course of lysosomal drug uptake monitored by hsSRS imaging.
Figure 6: Intracellular interaction of TKI drugs with chloroquine measured by hsSRS imaging.

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Acknowledgements

We thank S. Martin, J. Hastewell, M. Ji, F-K. Lu, C. Freudiger and W. Yang for helpful discussions. We also thank S. Moss for his help with the Raman peak assignments. This work was supported by the National Institute of Health's T-R01 (1R01EB010244-01) awarded to X.S.X.

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  1. Dan Fu and Jing Zhou: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA

    Dan Fu & X. Sunney Xie

  2. Novartis Institute for Biomedical Research, Cambridge, Massachusetts 02139, USA

    Jing Zhou, Wenjing Suzanne Zhu, Y. Karen Wang, Tami Hood & Andrew Wylie

  3. Novartis Institute for Biomedical Research, Basel CH-4002, Switzerland

    Paul W. Manley

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Contributions

D.F., J.Z., Y.K.W. and X.S.X conceived the study. D.F. and J.Z. designed the study. D.F., J.Z., Y.K.W., X.S.X., and P.W.M participated extensively in the scientific discussion about the study. D.F. performed the hsSRS imaging study and analysed the imaging data. W.S.Z. performed the cell proliferation and phospho-STAT5 assay, with supervision from A.W., and J.Z., W.S.Z., A.W. and T.H. prepared the drug and cell samples. D.F., J.Z., P.M. and X.S.X. wrote the manuscript with contributions from W.S.Z., A.W. and T.H.

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Correspondence to X. Sunney Xie.

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Fu, D., Zhou, J., Zhu, W. et al. Imaging the intracellular distribution of tyrosine kinase inhibitors in living cells with quantitative hyperspectral stimulated Raman scattering. Nature Chem 6, 614–622 (2014). https://doi.org/10.1038/nchem.1961

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