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Intracerebral fate of organic and inorganic nanoparticles is dependent on microglial extracellular vesicle function

Nature Nanotechnology volume 19, pages 376–386 (2024)Cite this article

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Abstract

Nanoparticles (NPs) represent an important advance for delivering diagnostic and therapeutic agents across the blood–brain barrier. However, NP clearance is critical for safety and therapeutic applicability. Here we report on a study of the clearance of model organic and inorganic NPs from the brain. We find that microglial extracellular vesicles (EVs) play a crucial role in the clearance of inorganic and organic NPs from the brain. Inorganic NPs, unlike organic NPs, perturb the biogenesis of microglial EVs through the inhibition of ERK1/2 signalling. This increases the accumulation of inorganic NPs in microglia, hindering their elimination via the paravascular route. We also demonstrate that stimulating the release of microglial EVs by an ERK1/2 activator increased the paravascular glymphatic pathway-mediated brain clearance of inorganic NPs. These findings highlight the modulatory role of microglial EVs on the distinct patterns of the clearance of organic and inorganic NPs from the brain and provide a strategy for modulating the intracerebral fate of NPs.

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Fig. 1: Inorganic and organic NPs exhibit different brain distributions and clearance profiles.
Fig. 2: Inorganic and organic NPs manifest different intracellular transport and exocytosis patterns in microglia.
Fig. 3: NP treatment alters microglial signalling and exocytosis processes.
Fig. 4: Microglial EV release is crucial to NP elimination.
Fig. 5: ERK1/2 activation facilitates NP clearance from the brain.

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Data availability

All data supporting the findings of this study are available within the Article and the Supplementary Information. All RNA sequence raw data were deposited to the NCBI Sequence Read Archive (https://www.ncbi.nlm.nih.gov/sra) with the identifier BioProject ID: PRJNA1003397. Proteomics raw data have been deposited to the PRIDE archive (https://www.ebi.ac.uk/pride/archive) with the identifier PXD044395. Source data are provided with this paper.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (81973272, 92068111, 82073836, 82171358), the National Key Research and Development Program of China (2022YFC2502800), the Canada Research Chairs Program (950-232468) and grants from the Shanghai Science and Technology Committee (21XD1422200, 23S41900100) and the Innovation Program of Shanghai Municipal Education Commission (2023ZKZD21). We thank Y. Wu, G. Li and Y. Huang from the Core Facility of Basic Medical Sciences, Shanghai Jiao Tong University School of Medicine. We are also grateful to J. Chen from the Instrumental Analysis Center of Shanghai Jiao Tong University for providing instrumental assistance.

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Authors and Affiliations

  1. Department of Pharmacology and Chemical Biology, Shanghai Universities Collaborative Innovation Center for Translational Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Jinchao Gao, Qingxiang Song, Xiao Gu, Gan Jiang, Jialin Huang, Yuyun Tang, Renhe Yu, Antian Wang, Yukun Huang & Xiaoling Gao

  2. Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada

    Gang Zheng

  3. Shuguang Lab for Future Health, Academy of Integrative Medicine, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China

    Hongzhuan Chen

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Contributions

J.G. and X. Gao are responsible for all phases of the research. J.G. designed and executed the experiments, analysed the data and wrote the original manuscript. X. Gao conceptualized and supervised the project and contributed to the experimental planning, data analysis and manuscript revision. H.C. and G.Z. conceptualized and supervised the project and contributed to the manuscript revision. Q.S. and X. Gu helped in designing and executing the in vivo two-photon laser-scanning microscopy experiments. G.J., J.H. and A.W. helped with the preparation of organic nanoparticles. Y.T. helped with the extraction of exosomes. R.Y. and Y.H. helped with in vitro BV-2 cell studies. All authors reviewed and approved the paper.

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Correspondence to Gang Zheng, Hongzhuan Chen or Xiaoling Gao.

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Nature Nanotechnology thanks Elizabeth Nance and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–29, Discussion and Tables 1–6.

Reporting Summary

Supplementary Video 1

DiD-rHDL uptake (0–1 h). BV-2 cells were exposed to DMEM with 1% DiD-rHDL (25 μg/ml) for 1 h. Cells were stained with Hoechst 33342 (blue).

Supplementary Video 2

DiD-PEG-PLA NPs uptake (0–1 h). BV-2 cells were exposed to DMEM with 1% DiD-PEG-PLA (50 μg/ml) for 1 h. Cells were stained with Hoechst 33342 (blue).

Supplementary Video 3

QD uptake (0–1 h). BV-2 cells were exposed to DMEM with QD (30 μg/ml) for 1 h. Cells were stained with Hoechst 33342 (blue).

Supplementary Video 4

PQD uptake (0–1 h). BV-2 cells were exposed to DMEM with PQD (30 μg/ml) for 1 h. Cells were stained with Hoechst 33342 (blue).

Supplementary Video 5

DiD-rHDL exocytosis (1–4 h). BV-2 cells were exposed to DMEM with 1% DiD-rHDL (25 μg/ml) (red) for 1 h. Then cells were gently washed twice with the fresh medium and incubated in the fresh culture medium for exocytosis for 3 h. BV-2 cells were stained with Lyso-Tracker (green) and Hoechst 33342 (blue).

Supplementary Video 6

DiD-PEG-PLA NPs exocytosis (1–4 h). BV-2 cells were exposed to DMEM with 1% DiD-PEG-PLA (50 μg/ml) (red) for 1 h. Then cells were gently washed twice with the fresh medium and incubated in the fresh culture medium for exocytosis for 3 h. BV-2 cells were stained with Lyso-Tracker (green) and Hoechst 33342 (blue).

Supplementary Video 7

QD exocytosis (1–4 h). BV-2 cells were exposed to DMEM with QD (30 μg/ml) (red) for 1 h. Then cells were gently washed twice with the fresh medium and incubated in the fresh culture medium for exocytosis for 3 h. BV-2 cells were stained with Lyso-Tracker (green) and Hoechst 33342 (blue).

Supplementary Video 8

PQD exocytosis (1–4 h). BV-2 cells were exposed to DMEM with PQD (30 μg/ml) (red) for 1 h. Then cells were gently washed twice with the fresh medium and incubated in the fresh culture medium for exocytosis for 3 h. BV-2 cells were stained with Lyso-Tracker (green) and Hoechst 33342 (blue).

Supplementary Data 1

Statistical source data of Supplementary Figures.

Supplementary Data 2

Unprocessed western blots for Supplementary Figures.

Source data

Source Data Fig. 1

Statistical source data.

Source Data Fig. 2

Statistical source data.

Source Data Fig. 3

Statistical source data.

Source Data Fig. 4

Statistical source data.

Source Data Fig. 5

Statistical source data.

Source Data Fig. 3

Unprocessed western blots.

Source Data Fig. 4

Unprocessed western blots.

Source Data Fig. 5

Unprocessed western blots.

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Gao, J., Song, Q., Gu, X. et al. Intracerebral fate of organic and inorganic nanoparticles is dependent on microglial extracellular vesicle function. Nat. Nanotechnol. 19, 376–386 (2024). https://doi.org/10.1038/s41565-023-01551-8

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