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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. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution

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subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS MICROENVIRONMENT-TRIGGERED MULTIMODAL PRECISION DIAGNOSTICS Article 15 July 2021 RENAL

CLEARABLE POLYFLUOROPHORE NANOSENSORS FOR EARLY DIAGNOSIS OF CANCER AND ALLOGRAFT REJECTION Article 14 April 2022 AND-GATED PROTEASE-ACTIVATED NANOSENSORS FOR PROGRAMMABLE DETECTION OF

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engineering of magnetic nanoworms for _in vivo_ tumor targeting. _Small_ 5, 694–700 (2009). Article  CAS  Google Scholar  Download references 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. AUTHOR INFORMATION Author notes * Ester J. Kwon and Jaideep S. Dudani: These authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Koch Institute for Integrative

Cancer Research, Massachusetts Institute of Technology (MIT), Cambridge, 02139, Massachusetts, USA. Ester J. Kwon, Jaideep S. Dudani & Sangeeta N. Bhatia * 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 * Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA. Jaideep S. Dudani * Department of Electrical Engineering and

Computer Science, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA. Sangeeta N. Bhatia * Department of Medicine, Brigham and Women’s Hospital and Harvard Medical

School, Boston, 02115, Massachusetts, USA. Sangeeta N. Bhatia * Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, 02139, Massachusetts, USA. Sangeeta N. Bhatia

* Howard Hughes Medical Institute, Cambridge, 02139, Massachusetts, USA. Sangeeta N. Bhatia Authors * Ester J. Kwon View author publications You can also search for this author inPubMed 

Google Scholar * Jaideep S. Dudani View author publications You can also search for this author inPubMed Google Scholar * Sangeeta N. Bhatia View author publications You can also search for

this author inPubMed Google Scholar CONTRIBUTIONS 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. CORRESPONDING AUTHOR Correspondence to Sangeeta N. Bhatia. ETHICS DECLARATIONS COMPETING INTERESTS The authors are listed as inventors on patent applications related to the

content of this work. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary text, figures and references. (PDF 12491 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT

THIS ARTICLE CITE THIS ARTICLE 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 Download citation * Received: 14 August 2016 * Accepted: 01 March 2017 * Published: 10 April 2017 * DOI: https://doi.org/10.1038/s41551-017-0054 SHARE

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