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Ultrasensitive tumour-penetrating nanosensors of protease activity

Nature Biomedical Engineering volume 1, Article number: 0054 (2017) Cite this article

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Abstract

The ability to identify cancer lesions with endogenous biomarkers is currently limited to tumours ~1 cm in diameter. We recently reported an exogenously administered tumour-penetrating nanosensor that sheds, in response to tumour-specific proteases, peptide fragments that can then be detected in the urine. Here, we report the optimization, informed by a pharmacokinetic mathematical model, of the surface presentation of the peptide substrates to both enhance on-target protease cleavage and minimize off-target cleavage, and of the functionalization of the nanosensors with tumour-penetrating ligands that engage active trafficking pathways to increase activation in the tumour microenvironment. The resulting nanosensor discriminated sub-5 mm lesions in human epithelial tumours and detected nodules with median diameters smaller than 2 mm in an orthotopic model of ovarian cancer. We also demonstrate enhanced receptor-dependent specificity of signal generation in the urine in an immunocompetent model of colorectal liver metastases, and in situ activation of the nanosensors in human tumour microarrays when re-engineered as fluorogenic zymography probes.

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Figure 1: MMP9 is upregulated across human cancers.
Figure 2: In vitro and in vivo experiments and in silico evaluation enables engineering of an optimized ABN.
Figure 3: Optimized ABNs detect sub-5 mm diameter tumours.
Figure 4: Urinary biomarkers outperform blood biomarkers in detecting millimetre-sized lesions in orthotopic xenograft models of ovarian cancer.
Figure 5: Minimally invasive receptor classification of syngeneic liver metastasis via targeted ABNs.
Figure 6: ABN zymography is responsive to human tissues.

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Acknowledgements

We thank H. Fleming (MIT) for critical editing of the manuscript, and A. Warren (MIT), G. Kwong (Georgia Institute of Technology), J. Voog (MIT, Massachusetts General Hospital), V. Ramanan and C. Buss (MIT) for helpful discussion. We are grateful to the Koch Institute Swanson Biotechnology Core at MIT, especially S. Malstrom, K. Cormier and veterinary pathologist R. T. Bronson. This study was supported in part by a Koch Institute Support Grant P30-CA14051 from the National Cancer Institute (Swanson Biotechnology Center), a Core Center Grant P30-ES002109 from the National Institute of Environmental Health Sciences, the Ludwig Fund for Cancer Research, and the Koch Institute Marble Center for Cancer Nanomedicine. E.J.K. acknowledges support from the Ruth L. Kirschstein National Research Service Award (1F32CA177094-01). J.S.D. thanks the National Science Foundation Graduate Research Fellowship Program for support. S.N.B. is a Howard Hughes Institute Investigator.

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Author notes

  1. Ester J. Kwon and Jaideep S. Dudani: These authors contributed equally to this work.

Authors and Affiliations

  1. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology (MIT), Cambridge, 02139, Massachusetts, USA.

    Ester J. Kwon, Jaideep S. Dudani & Sangeeta N. Bhatia

  2. Harvard–MIT Division of Health Sciences and Technology, Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA.

    Ester J. Kwon & Sangeeta N. Bhatia

  3. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA.

    Jaideep S. Dudani

  4. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA.

    Sangeeta N. Bhatia

  5. Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School, Boston, 02115, Massachusetts, USA.

    Sangeeta N. Bhatia

  6. Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, 02139, Massachusetts, USA.

    Sangeeta N. Bhatia

  7. Howard Hughes Medical Institute, Cambridge, 02139, Massachusetts, USA.

    Sangeeta N. Bhatia

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  1. Ester J. Kwon
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E.J.K. and J.S.D. performed the experiments and analysed the data. E.J.K., J.S.D. and S.N.B. designed the experiments and wrote the manuscript.

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Correspondence to Sangeeta N. Bhatia.

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Kwon, E., Dudani, J. & Bhatia, S. Ultrasensitive tumour-penetrating nanosensors of protease activity. Nat Biomed Eng 1, 0054 (2017). https://doi.org/10.1038/s41551-017-0054

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