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A nanoparticle probe for the imaging of autophagic flux in live mice via magnetic resonance and near-infrared fluorescence

Nature Biomedical Engineering volume 6, pages 1045–1056 (2022)Cite this article

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Abstract

Autophagy—the lysosomal degradation of cytoplasmic components via their sequestration into double-membraned autophagosomes—has not been detected non-invasively. Here we show that the flux of autophagosomes can be measured via magnetic resonance imaging or serial near-infrared fluorescence imaging of intravenously injected iron oxide nanoparticles decorated with cathepsin-cleavable arginine-rich peptides functionalized with the near-infrared fluorochrome Cy5.5 (the peptides facilitate the uptake of the nanoparticles by early autophagosomes, and are then cleaved by cathepsins in lysosomes). In the heart tissue of live mice, the nanoparticles enabled quantitative measurements of changes in autophagic flux, upregulated genetically, by ischaemia–reperfusion injury or via starvation, or inhibited via the administration of a chemotherapeutic or the antibiotic bafilomycin. In mice receiving doxorubicin, pre-starvation improved cardiac function and overall survival, suggesting that bursts of increased autophagic flux may have cardioprotective effects during chemotherapy. Autophagy-detecting nanoparticle probes may facilitate the further understanding of the roles of autophagy in disease.

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Fig. 1: Characterization of the ADN and its in vitro validation.
Fig. 2: Specificity of ADN activation and its uptake in early autophagosomes.
Fig. 3: Imaging of cardiomyocyte autophagy ex vivo with ADN is both sensitive and specific.
Fig. 4: ADN detects a reduction in autophagy when H9C2 cardiomyocytes are exposed to Dox and a cytoprotective effect when autophagy is restored.
Fig. 5: Starvation before Dox challenge in mice restores basal levels of autophagy, attenuates apoptosis, enhances cardiac function and improves overall survival.
Fig. 6: In vivo imaging of autophagy with ADN.

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Data availability

The main data supporting the results in this study are available within the paper and its Supplementary Information. The raw and analyzed datasets generated during the study are too large to be publicly shared but are available for research purposes from the corresponding authors on reasonable request. Source data are provided with this paper.

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Acknowledgements

This study was supported in part by the following grants from the National Institutes of Health: K99/R00HL121152 (to H.H.C.), R01HL112831 and R01HL141563 (to D.E.S.), R01HL122547 and R01HL102368-06A1 (to S.D.), R01HL131635 (to C.M.) and R01HL131831 (to R.M.B.). Electron microscopy was performed in the Microscopy Core of the Center for Systems Biology/Program in Membrane Biology, which is partially supported by an Inflammatory Bowel Disease Grant (DK043351) and a Boston Area Diabetes and Endocrinology Research Center (BADERC) award (DK057521). The funders had no role in the design of the study, in data collection and analysis, in the decision to publish the manuscript or in its preparation.

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Authors and Affiliations

  1. Molecular Cardiology Research Institute, Tufts Medical Center, Tufts University School of Medicine, Boston, MA, USA

    Howard H. Chen, Zehedina Khatun, Lan Wei, Asma Boukhalfa, Efosa Enoma, Lin Meng & Robert M. Blanton

  2. A.A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Howard H. Chen, Choukri Mekkaoui, Yinching I. Chen, Anand T. N. Kumar & David E. Sosnovik

  3. Cardiovascular Research Center, Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Dakshesh Patel, Sally Ji Who Kim, Leena Kaikkonen, Guoping Li, Parul Sahu, Saumya Das & David E. Sosnovik

  4. Program in Membrane Biology and Division of Nephrology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA

    Diane E. Capen

  5. Gordon Center for Medical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Hushan Yuan & Lee Josephson

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Contributions

H.H.C. and D.E.S. conceived the study, designed the experiments and performed data acquisition, data analysis, data interpretation, figure preparation and manuscript writing. Z.K. and L.W. contributed to data acquisition and data analysis. C.M. and S.J.W.K. contributed to data analysis and figure preparation. D.P., A.B., E.E., L.M., Y.I.C., L.K., G.L., D.E.C., P.S. and A.T.N.K. contributed to data acquisition and data analysis. R.M.B. contributed to data acquisition, data analysis and data interpretation. H.Y. contributed to experimental design, data acquisition and data analysis. S.D. contributed to experimental design, data analysis and data interpretation. L.J. contributed to the conception of the study and to experimental design and data interpretation. All authors contributed to the editing of the manuscript.

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Correspondence to Howard H. Chen.

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Chen, H.H., Khatun, Z., Wei, L. et al. A nanoparticle probe for the imaging of autophagic flux in live mice via magnetic resonance and near-infrared fluorescence. Nat. Biomed. Eng 6, 1045–1056 (2022). https://doi.org/10.1038/s41551-022-00904-3

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