Renal clearable catalytic gold nanoclusters for in vivo disease monitoring

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Abstract

Ultrasmall gold nanoclusters (AuNCs) have emerged as agile probes for in vivo imaging, as they exhibit exceptional tumour accumulation and efficient renal clearance properties. However, their intrinsic catalytic activity, which can enable an increased detection sensitivity, has yet to be explored for in vivo sensing. By exploiting the peroxidase-mimicking activity of AuNCs and the precise nanometre-size filtration of the kidney, we designed multifunctional protease nanosensors that respond to disease microenvironments to produce a direct colorimetric urinary readout of the disease state in less than one hour. We monitored the catalytic activity of AuNCs in the collected urine of a mouse model of colorectal cancer in which tumour-bearing mice showed a 13-fold increase in colorimetric signal compared to healthy mice. The nanosensors were eliminated completely through hepatic and renal excretion within four weeks of injection with no evidence of toxicity. We envision that this modular approach will enable the rapid detection of a diverse range of diseases by exploiting their specific enzymatic signatures.

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Fig. 1: Design of the nanocatalyst signal amplification sensing system.
Fig. 2: Peptide-functionalized AuNCs exhibit robust catalytic activity.
Fig. 3: Peptide-functionalized AuNCs clear via the renal system and retain catalytic activity in urine.
Fig. 4: AuNC–NAv complexes disassemble in vitro in response to protease activity.
Fig. 5: AuNC-functionalized protease nanosensors enable a direct colorimetric urinary readout of the disease state.

Data availability

Research data is available online at https://doi.org/10.5281/zenodo.3256265.

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Acknowledgements

We thank H. Fleming (MIT) and A. Nogiwa-Valdez (Imperial) for critical reading and editing of the manuscript, M. Kumar (Bioimaging and Chemical Analysis Facilities, MIT) for help with the ICP–MS, M. Berne (Tufts University Core Facility) for peptide synthesis and K. Cormier and R. Bronson from the Koch Institute Histology Core. We acknowledge use of the characterization facilities at the Harvey Flower Electron Microscopy Suite (Department of Materials, Imperial College London) and the Light Microscopy Facilities at the Francis Crick Institute London. 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. C.N.L., Q.C. and M.M.S. acknowledge generous support from the i-sense Engineering and Physical Sciences Research Council (EPSRC) IRC in Early Warning Sensing Systems for Infectious Diseases (EP/K031953/1; www.i-sense.org.uk). C.N.L. acknowledges support from the Marshall Aid Commemoration Commission. A.P.S. acknowledges support from the NIH Molecular Biophysics Training Grant and the National Science Foundation Graduate Research Fellowship. J.S.D. acknowledges support from the National Science Foundation Graduate Research Fellowship, the Ludwig Center for Molecular Oncology fellowship and the Siebel Scholar Foundation. A.N. acknowledges support from the Sir Henry Wellcome Postdoctoral Fellowship (209121_Z_17_Z) funding scheme from the Wellcome Trust. A.B. acknowledges support from the Early Postdoc Fellowship program (P2ELP2_178238) from the Swiss National Science Foundation. M.M.S., Y.L. and Q.C. acknowledge support from the European Research Council (ERC) Seventh Framework Programme Consolidator grant “Naturale CG” (616417). S.N.B. is a Howard Hughes Medical Institute Investigator.

Author information

C.N.L., A.P.S., J.S.D., S.N.B. and M.M.S. conceived and designed the research. C.N.L. and A.P.S. carried out all the experiments and analysed the data. Y.L. assisted with the peptide synthesis and characterization, A.N. performed the FCS measurements and analysis, A.B. assisted with the ICP–MS and Q.C. assisted with the TEM imaging. C.N.L., A.P.S., S.N.B. and M.M.S. wrote the manuscript with feedback from all the authors.

Correspondence to Sangeeta N. Bhatia or Molly M. Stevens.

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Competing interests

S.N.B., M.M.S., C.N.L. and A.P.S. have filed a patent application related to this research with the US Patent and Trademark Office. S.N.B. is a director at Vertex, co-founder and consultant at Glympse Bio, consultant for Cristal, Maverick and Moderna, and receives sponsored research funds from Johnson & Johnson.

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Peer review information: Nature Nanotechnology thanks Honggang Cui, Hui Wei and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Materials and Methods, Figs. 1–20, Tables 1–4 and refs. 1–3.

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