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Prodrug-tethered lipid nanoparticles for synergistic messenger RNA cancer immunotherapy

Nature Nanotechnology volume 21, pages 430–442 (2026) Cite this article

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Abstract

Regulating T cell phenotypes between activation and exhaustion remains a significant challenge for messenger RNA-based cancer immunotherapy. A potential approach to improve anti-cancer T cell activity is to co-deliver interleukin-12 (IL-12), to stimulate effector T cells, and indoleamine 2,3-dioxygenase (IDO) inhibitor, to suppress T cell exhaustion. Here we design prodrug ionizable lipid nanoparticles (pLNPs), via a library of prodrug ionizable lipids (pILs), incorporating an intracellularly cleavable IDO inhibitor within the pIL structure and encapsulating IL-12 messenger RNA. The lead pIL shows enhanced mRNA transfection over a clinically utilized ionizable lipid, as well as strong immunomodulatory effects via release of the IDO inhibitor. In a subcutaneous colon cancer mouse model, pLNP drives complete regression of primary tumours by eliciting effector T cell infiltration while reducing exhaustion, induces a memory T cell response and stimulates a systemic immune response that allows for regression of distal tumours in this study. These results highlight the promise of pLNPs for small-molecule drug and mRNA combination cancer immunotherapy.

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Fig. 1: Engineering IDO inhibitor-based prodrug ionizable lipids (pILs) to facilitate IL-12 mRNA delivery via prodrug lipid nanoparticles (pLNPs) for synergistic cancer immunotherapy.
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Fig. 2: In vitro and in vivo screening of pIL library and lead pLNPs for mRNA delivery.
The alternative text for this image may have been generated using AI.
Fig. 3: G0-SS-AA-C12 LNP for potent IL-12 mRNA transfection and suppression of T cell exhaustion in vitro.
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Fig. 4: G0-SS-AA-C12 IL-12 pLNP achieves synergistic therapeutic effects in vivo in an MC38 tumour model and induces a splenic T cell response.
The alternative text for this image may have been generated using AI.
Fig. 5: Characterization of TME after treatment with G0-SS-AA IL-12 pLNPs encapsulating IL-12 mRNA.
The alternative text for this image may have been generated using AI.
Fig. 6: Intratumoural injection of G0-SS-AA-C12 IL-12 pLNP elicits effective long-term anti-tumour response and drives regression of distal tumours.
The alternative text for this image may have been generated using AI.

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Data availability

All relevant data supporting the findings of this study are available within the paper, Supplementary Information or Source Data file. Source data are provided with this paper.

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Acknowledgements

M.J.M. acknowledges support from a Burroughs Wellcome Fund Career Award at the Scientific Interface (CASI), a US National Science Foundation CAREER award (CBET-2145491) and an American Cancer Society Research Scholar Grant (RSG-22-122-01-ET).

Author information

Author notes

  1. These authors contributed equally: Qiangqiang Shi, Ningqiang Gong.

Authors and Affiliations

  1. Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA

    Qiangqiang Shi, Ningqiang Gong, Jinjin Wang, Rohan Palanki, Zhangyi Luo, Junchao Xu, Christian G. Figueroa-Espada, Lulu Xue, Ye Zeng, Xuexiang Han, Dongyoon Kim, Hannah Yamagata, Hannah C. Geisler, Il-Chul Yoon & Michael J. Mitchell

  2. Department of General Surgery, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, Hefei National Research Center for Physical Science at Microscale, University of Science and Technology of China, Hefei, China

    Ningqiang Gong

  3. Department of Radiation Oncology, University of Pennsylvania, Philadelphia, PA, USA

    Qiuxian Zheng

  4. Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Mohamad-Gabriel Alameh, Garima Dwivedi, Benjamin Davis, Jilian Melamed, Rakan El-Mayta, Drew Weissman & Michael J. Mitchell

  5. School of Dental Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Qinyuan Chen

  6. Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Drew Weissman & Michael J. Mitchell

  7. Penn Institute for RNA Innovation, University of Pennsylvania, Philadelphia, PA, USA

    Michael J. Mitchell

  8. Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Michael J. Mitchell

  9. Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Michael J. Mitchell

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  1. Qiangqiang Shi
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Contributions

Q.S., N.G. and M.J.M. conceived and designed the experiments. Q.S., N.G., J.W., Q.Z., M.-G.A., G.D., B.D., J.M., Z.L., R.P., J.X., Y.Z., L.X., X.H., D.K., Q.C., H.Y., H.C.G., R.E.-M. and I.-C.Y. performed the experiments. Q.S., N.G. and M.J.M. analysed the data. Q.S., R.P., C.G.F.-E., H.C.G., D.W. and M.J.M. wrote, reviewed and edited the paper. M.J.M. supervised the entire project. All authors discussed the results and commented on the paper.

Corresponding author

Correspondence to Michael J. Mitchell.

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Competing interests

M.J.M. and Q.S. have filed a patent application based on this study (Application No. PCT/US25/52032). The other authors declare no competing interests.

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Nature Nanotechnology thanks Philippe Barthélémy and Quanyin Hu for their contribution to the peer review of this work.

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Shi, Q., Gong, N., Wang, J. et al. Prodrug-tethered lipid nanoparticles for synergistic messenger RNA cancer immunotherapy. Nat. Nanotechnol. 21, 430–442 (2026). https://doi.org/10.1038/s41565-025-02102-z

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