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[18F]-C-SNAT4: an improved caspase-3-sensitive nanoaggregation PET tracer for imaging of tumor responses to chemo- and immunotherapies

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Abstract

Positron emission tomography (PET) imaging of apoptosis can noninvasively detect cell death in vivo and assist in monitoring tumor response to treatment in patients. While extensive efforts have been devoted to addressing this important need, no apoptosis PET imaging agents have yet been approved for clinical use. This study reports an improved 18F-labeled caspase-sensitive nanoaggregation tracer ([18F]-C-SNAT4) for PET imaging of tumor response to chemo- and immunotherapies in preclinical mouse models.

Methods

We rationally designed and synthesized a new PET tracer [18F]-C-SNAT4 to detect cell death both in vitro and in vivo. In vitro radiotracer uptake studies were performed on drug-sensitive and -resistant NSCLC cell lines (NCI-H460 and NCI-H1299, respectively) treated with cisplatin at different doses. In vivo therapy response monitoring by [18F]-C-SNAT4 PET imaging was evaluated with two treatment modalities—chemotherapy and immunotherapy in two tumor xenografts in mice. Radiotracer uptake in the tumors was validated ex vivo using γ-counting and cleaved caspase-3 immunofluorescence.

Results

This [18F]-C-SNAT4 PET tracer was facilely synthesized and displayed improved serum stability profiles. [18F]-C-SNAT4 cellular update was elevated in NCI-H460 cells in a time- and dose-dependent manner, which correlated well with cell death. A significant increase in [18F]-C-SNAT4 uptake was measured in NCI-H460 tumor xenografts in mice. In contrast, a rapid clearance of [18F]-C-SNAT4 was observed in drug-resistant NCI-H1299 in vitro and in tumor xenografts. Moreover, in BALB/C mice bearing murine colon cancer CT26 tumor xenografts receiving checkpoint inhibitors, [18F]-C-SNAT4 showed its ability for monitoring immunotherapy-induced apoptosis and reporting treatment-responding mice from non-responding.

Conclusion

The uptake of [18F]-C-SNAT4 in tumors received chemotherapy and immunotherapy is positively correlated with the tumor apoptotic level and the treatment efficacy. [18F]-C-SNAT4 PET imaging can monitor tumor response to two different treatment modalities and predict the therapeutic efficacy in preclinical mouse models.

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Data availability

Original data are available on request.

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Acknowledgements

This work was supported by a grant from National Cancer Institute (NCI) Stanford CCNE-TD (U54CA199075). M.C. acknowledges the support by the NCI-funded Stanford Cancer Translational Nanotechnology Training grant (T32CA196585). L.C. acknowledges the support by the Office of the Assistant Secretary of Defense for Health Affairs through the Breast Cancer Research Program (BCRP) under Award No. (W81XWH-18-1-0591). The authors acknowledge the use of Stanford Center for Innovation in In-Vivo Imaging (SCI3) Core Facility, Stanford Neuroscience Microscopy Service (Supported by NIH NS069375), and Stanford Nano Shared Facilities (SNSF) (Supported by the National Science Foundation under award ECCS-1542152). We thank Emily Johnson and Michelle James for their technical assistance in the PET experiments, and Xue Wu for her assistance in the biodistribution experiments.

Funding

This work was supported by a grant from National Cancer Institute (NCI) Stanford CCNE-TD (U54CA199075). M.C. acknowledges the support by the NCI-funded Stanford Cancer Translational Nanotechnology Training grant (T32CA196585). L.C. acknowledges the support by the Office of the Assistant Secretary of Defense for Health Affairs through the Breast Cancer Research Program (BCRP) under Award No. (W81XWH-18-1-0591). The authors acknowledge the use of Stanford Center for Innovation in In-Vivo Imaging (SCI3) Core Facility, Stanford Neuroscience Microscopy Service (Supported by NIH NS069375), and Stanford Nano Shared Facilities (SNSF) (Supported by the National Science Foundation under award ECCS-1542152).

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  1. Min Chen and Zixin Chen contributed equally to this work.

Authors and Affiliations

  1. Molecular Imaging Program at Stanford, Department of Radiology, Stanford University School of Medicine, 1201 Welch Rd., Rm.P078, Stanford, CA, 94305-5484, USA

    Min Chen, Jessa B. Castillo, Liyang Cui, Kaixiang Zhou, Bin Shen, Jinghang Xie, Frederick T. Chin & Jianghong Rao

  2. Department of Chemistry, Stanford University, Stanford, CA, 94305, USA

    Zixin Chen & Jianghong Rao

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  1. Min Chen
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Correspondence to Jianghong Rao.

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Conflict of interest

M. Chen, Z. Chen, J. Xie, and J. Rao are inventors of a patent application filed by Stanford University covering part of the information contained in the paper. No other potential conflicts of interest relevant to this article exist.

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Movie S1

Real-time PET imaging of a mouse with lung tumor NCI-H460 after treatment with cisplatin. (AVI 32765 kb)

Movie S2

Real-time PET imaging of a mouse with lung tumor NCI-H1299 after treatment with cisplatin. (AVI 43787 kb)

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Chen, M., Chen, Z., Castillo, J.B. et al. [18F]-C-SNAT4: an improved caspase-3-sensitive nanoaggregation PET tracer for imaging of tumor responses to chemo- and immunotherapies. Eur J Nucl Med Mol Imaging 48, 3386–3399 (2021). https://doi.org/10.1007/s00259-021-05297-0

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