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Photoacoustic imaging of elevated glutathione in models of lung cancer for companion diagnostic applications

Nature Chemistry volume 13, pages 1248–1256 (2021)Cite this article

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Abstract

Companion diagnostics (CDx) are powerful tests that can provide physicians with crucial biomarker information that can improve treatment outcomes by matching therapies to patients. Here, we report a photoacoustic imaging-based CDx (PACDx) for the selective detection of elevated glutathione (GSH) in a lung cancer model. GSH is abundant in most cells, so we adopted a physical organic chemistry approach to precisely tune the reactivity to distinguish between normal and pathological states. To evaluate the efficacy of PACDx in vivo, we designed a blind study where photoacoustic imaging was used to identify mice bearing lung xenografts. We also employed PACDx in orthotopic lung cancer and liver metastasis models to image GSH. In addition, we designed a matching prodrug, PARx, that uses the same SNAr chemistry to release a chemotherapeutic with an integrated PA readout. Studies demonstrate that PARx can inhibit tumour growth without off-target toxicity in a lung cancer xenograft model.

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Fig. 1: Application of physical organic chemistry to tune SNAr reactivity for GSH sensing.
Fig. 2: Evaluation of PACDx GSH-responsiveness in vitro and in in cellulo systems.
Fig. 3: Development of PARx, a gemcitabine-based prodrug with matching GSH reactivity.
Fig. 4: Assessment of PARx biodistribution and in vivo activation.
Fig. 5: Determination of PARx efficacy in a murine model of lung cancer.
Fig. 6: Application of PACDx and PARx in unbiased animal studies.

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All data are available within the Article and its Supplementary Information. Alternatively, data are available upon request from the corresponding author.

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Acknowledgements

This work was supported by the National Institutes of Health (R35GM133581). M.Y.L acknowledges the Alfred P. Sloan Foundation for financial support. Major funding for the 500-MHz Bruker CryoProbeTM was provided by the Roy J. Carver Charitable Trust (Muscatine, Iowa; grant no. 15-4521) to the School of Chemical Sciences NMR Lab. The Q-TOF Ultima mass spectrometer was purchased in part with a grant from the National Science Foundation, Division of Biological Infrastructure (DBI-0100085). We also acknowledge the Core Facilities at the Carl R. Woese Institute for Genomic Biology for access to the Zeiss LSM 700 confocal microscope and corresponding software. We also acknowledge I. Dobrucka and the Molecular Imaging Laboratory at the Beckman Institute for use of the IVIS imaging system. M.Y.L. thanks H. Knox for initial animal training and assistance with the NanoZoomer. We thank T. Bearrood and C. Anorma for assistance with initial confocal imaging experiments, L. Akin for aid with mass spectrometry experiments, S. Anakk and A. Dean for help with interpreting results from H&E staining experiments, N. Herndon and J. Xu for help with generating the orthotopic lung cancer and liver metastasis models, S. Gardner for providing 4T1 tumour models, J. Sarol and A. Kaur from Biostatistics, Epidemiology, & Research Design at the Interdisciplinary Health Sciences Institute (UIUC) for aid in statistical analysis, and A. Bennet for helpful discussions.

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  1. Department of Chemistry and Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    Melissa Y. Lucero & Jefferson Chan

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  1. Melissa Y. Lucero
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Contributions

M.Y.L. performed all experiments in this study that include chemical synthesis, in vitro characterization, cellular studies, tumour model studies, in vivo imaging and sample preparation for ex vivo analysis. J.C. assisted with the blinded animal study. M.Y.L. and J.C. analysed the data and prepared the manuscript. J.C. conceived the project, with intellectual contributions from M.Y.L.

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Correspondence to Jefferson Chan.

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Peer review information Nature Chemistry thanks Junjie Yao, Deju Ye and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Figs. 1–34, Tables 1 and 2 and NMR spectra 2–21.

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Lucero, M.Y., Chan, J. Photoacoustic imaging of elevated glutathione in models of lung cancer for companion diagnostic applications. Nat. Chem. 13, 1248–1256 (2021). https://doi.org/10.1038/s41557-021-00804-0

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