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Spatially resolved in vivo imaging of inflammation-associated mRNA via enzymatic fluorescence amplification in a molecular beacon

Nature Biomedical Engineering volume 6, pages 1074–1084 (2022)Cite this article

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Abstract

The in vivo optical imaging of RNA biomarkers of inflammation is hindered by low signal-to-background ratios, owing to non-specific signal amplification in healthy tissues. Here we report the design and in vivo applicability, for the imaging of inflammation-associated messenger RNAs (mRNAs), of a molecular beacon bearing apurinic/apyrimidinic sites, whose amplification of fluorescence is triggered by human apurinic/apyrimidinic endonuclease 1 on translocation from the nucleus into the cytoplasm specifically in inflammatory cells. We assessed the sensitivity and tissue specificity of an engineered molecular beacon targeting interleukin-6 (IL-6) mRNA in live mice, by detecting acute inflammation in their paws and drug-induced inflammation in their livers. This enzymatic-amplification strategy may enable the specific and sensitive imaging of other disease-relevant RNAs in vivo.

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Fig. 1: Imaging of inflammation-associated mRNA via enzymatic fluorescence amplification.
Fig. 2: Evaluation of APE1-activated signal-amplification strategy in test tubes.
Fig. 3: APE1 translocation-mediated imaging of IL-6 mRNA in inflammatory cells.
Fig. 4: Evaluation of the signal-amplification strategy for imaging of acute inflammation in vivo.
Fig. 5: Evaluation of the signal-amplification strategy for diagnosis of drug-induced hepatotoxicity in vivo.
Fig. 6: Comparison with clinical assays.

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 large to be publicly shared, yet they 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 work was supported financially by the National Natural Science Foundation of China (22125402 (to L.L.) and 22004023 (to J.Z.)), the Youth Innovation Promotion Association CAS (to J.Z.), the National Key R&D Program of China (2021YFA1200104, to L.L.) and the Strategic Priority Research Program of Chinese Academy of Sciences (XDB36000000, to L.L. and Y.Z.).

Author information

Author notes

  1. These authors contributed equally: Chuangui Sheng, Jian Zhao.

Authors and Affiliations

  1. CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety and CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology, Beijing, China

    Chuangui Sheng, Jian Zhao, Zhenghan Di, Yuliang Zhao & Lele Li

  2. College of Materials Science and Optoelectronic Technology, University of Chinese Academy of Sciences, Beijing, China

    Chuangui Sheng, Jian Zhao, Zhenghan Di, Yuliang Zhao & Lele Li

  3. Advanced Research Institute of Multidisciplinary Science, Beijing Institute of Technology, Beijing, China

    Yuanyu Huang

  4. GBA Research Innovation Institute for Nanotechnology, Guangdong, China

    Yuliang Zhao & Lele Li

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Contributions

C.S., J.Z. and L.L conceived the idea and designed all the experiments. C.S. and J.Z. performed the experiments. Y.H. provided the PEGylated tri-block co-polymer. C.S., J.Z., Z.D., Y.Z. and L.L. analysed and discussed the results. C.S., J.Z. and L.L. wrote the manuscript. All authors reviewed the manuscript and approved the final version.

Corresponding author

Correspondence to Lele Li.

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Nature Biomedical Engineering thanks Li-Qun Gu and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Sheng, C., Zhao, J., Di, Z. et al. Spatially resolved in vivo imaging of inflammation-associated mRNA via enzymatic fluorescence amplification in a molecular beacon. Nat. Biomed. Eng 6, 1074–1084 (2022). https://doi.org/10.1038/s41551-022-00932-z

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