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Non-invasive monitoring of chronic liver disease via near-infrared and shortwave-infrared imaging of endogenous lipofuscin

An Author Correction to this article was published on 24 November 2020

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Abstract

Monitoring the progression of non-alcoholic fatty liver disease is hindered by a lack of suitable non-invasive imaging methods. Here, we show that the endogenous pigment lipofuscin displays strong near-infrared and shortwave-infrared fluorescence when excited at 808 nm, enabling label-free imaging of liver injury in mice and the discrimination of pathological processes from normal liver processes with high specificity and sensitivity. We also show that the near-infrared and shortwave-infrared fluorescence of lipofuscin can be used to monitor the progression and regression of liver necroinflammation and fibrosis in mouse models of non-alcoholic fatty liver disease and advanced fibrosis, as well as to detect non-alcoholic steatohepatitis and cirrhosis in biopsied samples of human liver tissue.

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Fig. 1: Real-time and non-invasive imaging of quantifiable autofluorescent pigments in the livers of rodents with progressive fibrosis.
Fig. 2: Quantification of lipofuscin/ceroid pigment by autofluorescence microscopy in progressive liver fibrosis model tissue.
Fig. 3: Regression of liver fibrosis is accompanied by the reversal of the lipofuscin/ceroid autofluorescence signal.
Fig. 4: Tracking lipofuscin autofluorescence with the progression of NAFLD in a high-fat diet mouse model.
Fig. 5: Significant autofluorescence signal and fibrosis in a lipodystrophy mouse model.
Fig. 6: Liver injury in humans with restructuring of lipofuscin distribution and quantity.

Data availability

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

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Acknowledgements

We thank S. Roberge for excellent technical support, W. Ho for RT-qPCR support and T. Reiberger for early discussions. This work received financial support, in part, from the National Institutes of Health through the Laser Biomedical Research Center (grant no. 9-P41-EB015871-26A1 to M.G.B.) and the US Army Research Office through the Institute for Soldier Nanotechnologies at MIT, under cooperative agreement no. W911NF-18-2-0048 (to M.G.B.). M.S. was supported by the National Science Foundation Graduate Research Fellowship Program (grant no. 1122374). J.A.C. was supported by a National Defense Science and Engineering Graduate Fellowship 32 CFR 168a. K.v.L., Y.Z. and M.G.B. were supported by a Grand Challenges Grant from the MGH/MIT Strategic Initiative. This work was also in part supported by grants from the National Foundation for Cancer Research, the Ludwig Center at Harvard, the US National Cancer Institute (grant nos. P01-CA080124, R35-CA197743, R01-CA208205 and U01-CA224173 to R.K.J.). J.M.P. was supported by a NIH Training Grant (grant no. T32HL007627). This work was further supported in part by the NIH (grant nos. R01 DK031036 and R01 DK033201 to C.R.K., and grant nos. K12 HD000850 and P30 DK40561) and a NASPGHAN Young Investigator/Nestle Nutrition Award to S.S. M.P. was supported by an Erwin Schroedinger Fellowship from the Austrian Science Fund (FWF; project number J 3747-B28). S.C. was supported by an ABTA Basic Research Fellowship, MGH Tosteon and Fund for Medical Discovery Fellowship and PCRF Emerging Investigator Fellowship Grant. O.T.B. was supported by a European Molecular Biology Organization long-term fellowship.

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Contributions

Conceptualization/evolution: W.J.K., J.A.C., I.X.C., M.P., K.v.L., O.T.B., R.K.J. and M.G.B. Formal analysis: M.S., W.J.K., J.A.C. and I.X.C. Funding acquisition: S.S., C.R.K., K.v.L., R.K.J. and M.G.B. Investigation/experiments: M.S., W.J.K., J.A.C., I.X.C., J.M.P., J.Z., Y.Z., M.P. and S.C. Methodology: M.S., W.J.K., J.A.C., I.X.C., J.M.P., A.S., R.K.J. and M.G.B. Project management: M.S., W.J.K. and J.A.C. Resources: S.S., C.R.K., K.v.L., R.K.J. and M.G.B. Supervision: R.K.J. and M.G.B. Validation: M.S., W.J.K. and J.A.C. Visualization: M.S. and J.A.C. Writing of the original draft: M.S., W.J.K. and J.A.C. Writing, review and editing: all authors.

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Correspondence to Rakesh K. Jain or Moungi G. Bawendi.

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Competing interests

M.P. is an investigator for Bayer, BMS and Lilly; a consultant for Bayer, BMS, Ipsen, Eisai and Lilly; and has received speaker honoraria from Bayer, BMS, Eisai and MSD as well as travel support from Bayer and BMS. R.K.J. received honorarium from Amgen; consultant fees from Chugai, Merck, Ophthotech, Pfizer, SPARC, SynDevRx and XTuit; owns equity in Enlight, Ophthotech, SynDevRx; and serves on the Boards of Trustees of Tekla Healthcare Investors, Tekla Life Sciences Investors, Tekla Healthcare Opportunities Fund and Tekla World Healthcare Fund. M.G.B. is a consultant and owns equity in Lumicell Inc. Neither any reagent nor any funding from these organizations was used in this study. MIT and MGH have filed a patent (United States application no. 62/654,665; international application no. PCT/US2019/026550) based on some of the findings described in this manuscript.

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Supplementary Dataset 1

Supplementary Tables 1–11.

Supplementary Dataset 2

Statistical details.

Supplementary Video 1

Real-time and non-invasive imaging of quantifiable autofluorescent pigments in rodent liver with progressive fibrosis.

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Saif, M., Kwanten, W.J., Carr, J.A. et al. Non-invasive monitoring of chronic liver disease via near-infrared and shortwave-infrared imaging of endogenous lipofuscin. Nat Biomed Eng 4, 801–813 (2020). https://doi.org/10.1038/s41551-020-0569-y

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