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Generation of tolerogenic antigen-presenting cells in vivo via the delivery of mRNA encoding PDL1 within lipid nanoparticles

Nature Biomedical Engineering volume 9, pages 1320–1334 (2025)Cite this article

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Abstract

Tolerogenic antigen-presenting cells (APCs) are promising as therapeutics for suppressing T cell activation in autoimmune diseases. However, the isolation and ex vivo manipulation of autologous APCs is costly, and the process is customized for each patient. Here we show that tolerogenic APCs can be generated in vivo by delivering, via lipid nanoparticles, messenger RNA coding for the inhibitory protein programmed death ligand 1. We optimized a lipid-nanoparticle formulation to minimize its immunogenicity by reducing the molar ratio of nitrogen atoms on the ionizable lipid and the phosphate groups on the encapsulated mRNA. In mouse models of rheumatoid arthritis and ulcerative colitis, subcutaneous delivery of nanoparticles encapsulating mRNA encoding programmed death ligand 1 reduced the fraction of activated T cells, promoted the induction of regulatory T cells and effectively prevented disease progression. The method may allow for the engineering of APCs that target specific autoantigens or that integrate additional inhibitory molecules.

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Fig. 1: In vivo-produced tol-APCs by LNPs/mPDL1 selectively target activated T cells for the treatment of AIDs.
Fig. 2: Optimization of LNP formulation with low immunogenicity using DOE.
Fig. 3: Production of tol-APCs by LNPs/mPDL1 treatment.
Fig. 4: Tol-APCs produced by LNPs/mPDL1 reduce proliferation and induce apoptosis of activated T cells in vitro.
Fig. 5: In vivo-produced tol-APCs by LNPs/mPDL1 treatment selectively reduce activated T cells in vivo.
Fig. 6: In vivo-produced tol-APCs inhibit the progression of RA.
Fig. 7: In vivo-produced tol-APCs mediate potent therapeutic effects in DSS-induced UC mice.

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Data availability

The 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 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 by the National Natural Science Foundation of China (52025036 to Y.W., 82173390 to M.L. and 52495014 to Y.W.), the National Key R&D Program of China (2020YFA0710700 and 2022YFC2303300 to Y.W.), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB0490000 and XDB0940303 to Y.W.), the Anhui Provincial Key Research and Development Project (2023s07020019 to Y.W.), the Anhui Provincial Major Science and Technology Project (202303a07020010 to Y.W.), the Anhui Provincial Natural Science Foundation (2408085J042 to M.L.), the project of collaborative innovation for colleges of Anhui province (GXXT-2022-063 to M.L.) and the USTC Research Funds of the Double First-Class Initiative (YD9100002054 to Y.W. and YD9110002021 to M.L.). This work was partially carried out at the USTC Center for Micro and Nanoscale Research and Fabrication. This work was partially carried out at the Instruments Center for Physical Science, University of Science and Technology of China.

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  1. These authors contributed equally: Yang Liu, Qian Liu, Baowen Zhang.

Authors and Affiliations

  1. Department of Radiology, the First Affiliated Hospital of University of Science and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China

    Yang Liu, Qian Liu, Shanshan Chen, Min Li & Yucai Wang

  2. National Key Laboratory of Immune Response and Immunotherapy, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China

    Yang Liu, Qian Liu, Baowen Zhang, Yanqiong Shen, Zhibin Li, Jiachen Zhang, Min Li & Yucai Wang

  3. Institute of Health and Medicine, Hefei Comprehensive National Science Center, Hefei, China

    Yanqiong Shen & Yucai Wang

  4. RNAlfa Biotech, Hefei, China

    Yanqiong Shen, Yi Yang & Yucai Wang

  5. Key Laboratory of Anhui Province for Emerging and Reemerging Infectious Diseases, Hefei, China

    Min Li & Yucai Wang

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Contributions

Y.W., M.L., Y.L. and Q.L. conceptualized and designed the research. Y.L., Q.L., B.Z., S.C., Y.S., Z.L., J.Z. and Y.Y. performed the experiments. S.C. provided help in designing LNP formulations. Y.L., Q.L. and B.Z. analysed the experimental data. Y.L., M.L., Q.L., B.Z. and Y.W. prepared the figures and wrote the paper. Y.W. supervised the project.

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Correspondence to Min Li or Yucai Wang.

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Extended data

Extended Data Fig. 1 In vivo-produced tol-APCs inhibit RA progression.

a, Statistical data of OARSI score. b, c, The percentage of IFN-γ+ (b) and TNF-α+ (c) area per FOV. d, Representative images of CD4, CD8, and Foxp3 staining from the knee joint of one mouse in a group of four. Scale bar = 200 µm. Arrows refer to Foxp3+ cells. e-g, Number of CD4+ (e), CD8+ (f) and Foxp3+ (g) cells per FOV. RA mice were subcutaneously treated with PBS, LNPs, or LNPs/mPDL1 (5 μg mRNA) at the lower right back. Mice treated with iTNF-α served as the positive control group. Normal group comprises healthy mice. n = 4 biologically independent mice per group for data in a-c and e-g. Data are expressed as the mean ± s.e.m. Statistical significances were determined using one-way ANOVA with Dunnett’s post hoc test. Comparisons were performed between the LNPs/mPDL1 group and each of the other groups. N.S. is P ≥ 0.05, and significant P values are displayed.

Source data

Extended Data Fig. 2 In vivo-produced tol-APCs mediate potent therapeutic effects in DSS-induced UC mice.

a, Representative images of CD8, Foxp3, IFN-γ, and TNF-α staining from the colon of one mouse in a group of four. Arrows refer to Foxp3+ cells. Scale bar = 200 µm. b-e, The number of CD8+ (b) and Foxp3+ (c) cells and the percentage of IFN-γ+ (d) and TNF-α+ (e) area per FOV. Mice were treated with PBS, LNPs, or LNPs/mPDL1 (5 μg mRNA) via subcutaneous injection at the lower right back. Mice treated with cyclosporine served as the positive control group. Normal group comprises healthy mice. n = 4 biologically independent mice per group for data in b-e. Data are expressed as the mean ± s.e.m. Statistical significances were determined using one-way ANOVA with Dunnett’s post hoc test. Comparisons were performed between the LNPs/mPDL1 group and each of the other groups. N.S. is P ≥ 0.05, and significant P values are displayed.

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Liu, Y., Liu, Q., Zhang, B. et al. Generation of tolerogenic antigen-presenting cells in vivo via the delivery of mRNA encoding PDL1 within lipid nanoparticles. Nat. Biomed. Eng 9, 1320–1334 (2025). https://doi.org/10.1038/s41551-025-01373-0

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