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Polymer–mRNA complexes for monocyte-trafficked, lymph node-targeted cancer vaccination

Nature Biomedical Engineering (2026) Cite this article

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Abstract

Lymph nodes are the primary sites where adaptive immunity is initiated, yet most messenger RNA cancer vaccines reach them inefficiently and instead accumulate in organs such as the liver, limiting therapeutic potency and increasing systemic toxicity. Here we developed a transferrin receptor-associating polyplex formed by electrostatic complexation of mRNA with low-molecular-weight polyethylenimine that had been chemically modified with cyclic disulfide monomers to enhance nucleic acid binding stability, enable thiol-based transferrin receptor engagement and reduce off-target liver uptake. After subcutaneous administration, these polyplexes activated innate immunity, rapidly recruited monocytes with high transferrin receptor expression and bound these cells through cyclic disulfide-mediated interactions. Monocytes then trafficked the vaccine to draining lymph nodes, where mRNA translation and antigen presentation occurred. Delivery of ovalbumin and interleukin 12 mRNA elicited strong antigen-specific cytotoxic T cell responses and inhibited melanoma progression and metastatic disease. Studies using Survivin and human papillomavirus antigens in distinct tumour models demonstrated broad applicability. This monocyte-driven lymph node-targeting strategy enables potent and selective delivery of mRNA cancer vaccines.

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Fig. 1: Design and characterization of TRAP-mNV for enhanced cytosolic mRNA delivery.
The alternative text for this image may have been generated using AI.
Fig. 2: Monocyte-driven, lymph node-specific mRNA delivery and transfection.
The alternative text for this image may have been generated using AI.
Fig. 3: Activation of the STING pathway by TRAP induces monocyte recruitment at the injection site.
The alternative text for this image may have been generated using AI.
Fig. 4: High endothelial venule-mediated recruitment of TRAP-mNV into the lymph nodes.
The alternative text for this image may have been generated using AI.
Fig. 5: TRAP-mNV drives monocyte-to-dendritic cell differentiation to promote immune responses.
The alternative text for this image may have been generated using AI.
Fig. 6: Therapeutic effect of TRAP-mNV vaccination on the B16F10 tumour model.
The alternative text for this image may have been generated using AI.
Fig. 7: Therapeutic efficacy of TRAP–mRNA vaccines on the TC-1 tumour model.
The alternative text for this image may have been generated using AI.

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Data availability

All data supporting the findings of this study are available within the Article, Supplementary Information and Source Data. Source data are provided with this paper. Additional data are available from the corresponding authors on reasonable request.

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Acknowledgements

We acknowledge financial support from National Natural Science Foundation of China (grant numbers U25A20260 and 52233007), Jiangsu Provincial Department of Science and Technology (grant numbers BG2025060 and BG2025053), Natural Science Foundation of Jiangsu Province (grant number SBK20250405084). We thank L. Wang and J. Yang for synthesizing mRNA, and J. Zhu for assistance in the preliminary cell experiments. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

Author information

Author notes

  1. These authors contributed equally: Qiongzhe Ren, Xiaofei Zhao, Lili Zhou.

Authors and Affiliations

  1. Biomedical Polymers Laboratory, College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou, P. R. China

    Qiongzhe Ren, Xiaofei Zhao, Ruonan Ye, Liguo Chen, Chao Deng, Fenghua Meng, Congcong Xu & Zhiyuan Zhong

  2. Institutes of Biology and Medical Science, Soochow University, Suzhou, P. R. China

    Lili Zhou & Fangfang Zhou

  3. Catug Biotechnology Co. Ltd, Suzhou, P. R. China

    Keyun Ren & Xijun Piao

  4. Department of Biosciences and Bioinformatics, School of Science, Xi’an Jiaotong–Liverpool University, Suzhou, P. R. China

    Yihan Zhou & Kevin C. Chan

  5. School of Pharmacy, Shanghai Jiao Tong University, Shanghai, P. R. China

    Yiming Qi

  6. College of Pharmaceutical Sciences, Soochow University, Suzhou, P. R. China

    Li Cao, Congcong Xu & Zhiyuan Zhong

  7. Suzhou Abogen Biosciences Co. Ltd, Suzhou, P. R. China

    Liang Du, Peng Gao & Bo Ying

  8. International College of Pharmaceutical Innovation, Soochow University, Suzhou, P. R. China

    Congcong Xu & Zhiyuan Zhong

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  1. Qiongzhe Ren
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  2. Xiaofei Zhao
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Contributions

Q.R., X.Z. and L.Z. performed experiments, analysed data and wrote the manuscript. R.Y., L.C., K.R., X.P., Y.Z., Y.Q., K.C.C., L.C., L.D. and P.G. assisted with the preparation of reagents, data analysis and manuscript preparation. Z.Z., C.X., F.Z., C.D., B.Y. and F.M. conceptualized the project, designed and supervised the research, and wrote the manuscript.

Corresponding authors

Correspondence to Fangfang Zhou, Congcong Xu or Zhiyuan Zhong.

Ethics declarations

Competing interests

Z.Z., C.X., R.Y., Q.R. and F.M. have filed a provisional patent (China, CN202510375982.0) related to DTC-modified PEI for mRNA delivery, which is relevant to the mRNA delivery platform investigated in this study. K.R. is a co-founder of Catug Biotechnology Co., Ltd, and X.P. is an employee of Catug Biotechnology Co., Ltd, which is involved in the synthesis of mRNA used in this work. B.Y. is the founder of Suzhou Abogen Biosciences, and L.D. and P.G. are employees of Suzhou Abogen Biosciences. The other authors declare no competing interests.

Peer review

Peer review information

Nature Biomedical Engineering thanks Lin Mei, Xianzhu Yang and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available.

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Extended data

Extended Data Fig. 1 Physicochemical characterization and transfection performance of TRAP–mRNA complexes.

a, Quantification of mRNA encapsulation efficiency by TRAP–mRNA and nTRAP–mRNA formulations (n = 6 independent measurements). b, Agarose gel electrophoresis showing that mRNA is effectively protected by TRAP. c, Time course of mRNA release from TRAP–mRNA complexes in the presence of GSH (0–24 h), as measured by a RibogreenTM assay (n = 3 independent measurements). d, Representative GFP protein fluorescence imaging of various cells, scale bar: 100 µm. e, Representative flow cytometry histograms of mEGFP expression in different cell lines. f, Quantification of GFP+ cells determined by flow cytometry (n = 3 biological samples). Data in a,c,f are presented as mean values ± s.d. The experiments in b and d were independently repeated at least three times with similar results. Statistical significance for panel f was determined by multiple two-sided unpaired Student’s t-tests with correction for multiple comparisons, by two-sided unpaired Student’s t-test for panel a. Exact P values are provided in the figures where applicable. Source data are provided.

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Extended Data Fig. 2 mRNA translation in lymph nodes following TRAP–mRNA administration.

a, Schematic illustration of Cre mRNA-mediated tdTomato expression in cells, where TRAP-mCre induces recombination and tdTomato expression through excision of the LoxP-flanked STOP cassette. b, Representative flow cytometry plots showing tdTomato+ DCs, Macs, and MOs in inguinal lymph nodes 48 h after subcutaneous injection of TRAP-mCre. c, Quantitative summary of Cre mRNA delivery efficiency, shown as the percentage of tdTomato+ cells among each immune-cell subset (n = 3 biological samples). d, Schematic representation of Thy1.1 (membrane-anchored) reporter expression following TRAP-mediated delivery of Thy1.1 mRNA, used to trace translation events in DCs and MOs within lymph nodes. e, Representative flow cytometry plots showing the frequency of Ly6C+CD11c+cells in inguinal lymph nodes at 3 h and 24 h after subcutaneous injection of TRAP–mLuc nanoparticles. f, Representative histogram overlays depicting Thy1.1 expression in Ly6C+CD11c− monocytes at 3 h, 12 h, and 24 h post-injection of TRAP–mThy1.1, and TRAP–mLuc treated mice served as isotype controls. Quantification of the MFI of Thy1.1+ MOs (n = 5 biological samples). Data in c and f are presented as mean values ± s.d. Statistical significance in f was determined by ordinary one-way ANOVA (two-sided). Source data are provided.

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Extended Data Fig. 3 TRAP enhances local immune-cell recruitment following subcutaneous administration.

a, Representative IVIS bioluminescence images showing luciferase expression after subcutaneous administration of TRAP–mLuc or non-targeted nTRAP–mLuc. b, Flow cytometry analysis of CD45+ immune-cell infiltration at the injection site in untreated (UT), nTRAP–mLuc, and TRAP–mLuc-treated mice. c, Quantification of CD45+ immune-cell proportions across groups (n = 5 biological samples). Data in c is presented as mean values ± s.d. Statistical significance in c was determined by ordinary one-way ANOVA (two-sided). Source data are provided.

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Extended Data Fig. 4 TRAP–mRNA uptake and monocyte differentiation dynamics at the injection site.

a, Subcutaneous administration of TRAP–mRNA leads to increased recruitment of CD11b+ myeloid cells at the injection site, as shown by representative flow cytometry histograms and quantification (n = 6 biological samples). b, Comparative analysis of TRAP–mRNA (Cy5-labelled) uptake between immune (CD45+) and non-immune (CD45−) cell populations at the injection site, shown by representative dot plots and fluorescence intensity histograms. c, Representative flow cytometry plot showing Cy5 signal in CD45+CD11b+ myeloid cells following subcutaneous injection of TRAP-Cy5-mRNA. d, Among Cy5+ myeloid cells, monocytes (Ly6C+), dendritic cells (CD11c+), and macrophages (F4/80+) account. e, Time-course analysis of monocyte differentiation at the injection site, as evidenced by the progressive increase in CD11c expression on Ly6C+ monocytes from 0 h to 8 h post-injection and quantification of Ly6C+CD11c+ double-positive cells over time (n = 5 biological samples). f, Temporal expression of XCR1 in Ly6C+ monocytes at the injection site, as assessed by flow cytometry at 3-, and 8-h post-injection, with representative plots and quantification (n = 5 biological samples). Data in a, e and f are presented as mean values ± s.d. Statistical significance was determined by one-way ANOVA (two-sided) for panels e and f, by two-sided unpaired Student’s t-test for panel a. Source data are provided.

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Extended Data Fig. 5 Systemic depletion of macrophages by clodronate liposome administration.

a, Schematic illustration of the in vivo macrophage depletion protocol. C57BL/6 mice were intraperitoneally injected with clodronate liposomes (CL-Lipo; 200 µl on day 0 and 100 µl on day 4), and tissues were harvested on day 5 for analysis. b–e, Representative flow cytometry plots and quantification of macrophages (F4/80+) and monocytes (Ly6C+) in (b) peripheral blood, (c) peritoneal lavage fluid, (d) spleens and (e) lymph nodes from untreated (UT) and CL-Lipo–treated mice (n = 5 mice per group). Data are presented as mean values ± s.d. Statistical significance was determined by a two-sided unpaired Student’s t-test. Exact P values are provided in the figure where applicable. Source data are provided.

Source data

Extended Data Fig. 6 Tracking in situ recruitment and TRAP-Thy1.1 mRNA expression in myeloid cells at the injection site.

a, Flow cytometry analysis of CD11b+ cells at 3 h, 12 h, and 24 h following subcutaneous injection of TRAP–mThy1.1. Representative gating on CD11b+ cells is shown, with quantification of DCs, Macs, and MOs among the recruited CD11b+ population at the injection site over time (n = 6 biological samples). b, Representative flow cytometry plots showing Thy1.1+ cells at 3 h, 12 h, and 24 h post-injection. Quantification of DCs, Macs, and MOs within the Thy1.1+ population reveals the distribution of translation-active myeloid subsets at the injection site (n = 6 biological samples). Thy1.1 encodes a membrane-anchored reporter protein, enabling detection of mRNA translation following TRAP-mediated delivery. Data in a and b are presented as mean values ± s.d. Statistical significance was determined by one-way ANOVA (two-sided). Exact P values are provided in the figure where applicable. Source data are provided.

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Extended Data Fig. 7 Comprehensive safety evaluation of TRAP–mRNA vaccination.

a, Complete blood count (CBC) parameters were analysed 24 h post-injection in mice receiving subcutaneous administration of TRAP–mRNA nanoparticles (3 µg mRNA, N/P = 10) or PBS (n = 6 mice per group), including WBC, NEU, LYM, RBC, HGB, HCT, PLT, MPV, NRBC, and ALY. No significant differences were observed between groups. b, Serum biochemical indicators of liver function (ALT and AST) measured 24 h post-injection (n = 4 mice per group) showed no evidence of hepatotoxicity in TRAP–mRNA-treated mice compared with PBS controls. c, Representative hematoxylin and eosin (H&E)-stained sections of major organs (heart, liver, spleen, lung, and kidney) collected 24 h post-injection from mice treated with TRAP–mLuc (control) or TRAP–mHPV revealed no apparent histopathological abnormalities, indicating minimal off-target tissue toxicity. Data are presented as mean ± s.d. Statistical significance was assessed using a two-tailed unpaired Student’s t-test. Exact P values are provided in the figure where applicable. Source data are provided.

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Supplementary information

Supplementary Information (download PDF )

Supplementary Table. 1, Supplementary Figs. 1–38 and Uncropped blots and gels of Supplementary Figs. 6a, 15 and 26c.

Reporting Summary (download PDF )

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Supplementary Video 1 (download GIF )

Cellular uptake of nanoparticles.

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Migration of nanoparticles within lymph nodes in the vasculature.

Supplementary Data 1 (download XLSX )

Coding sequences of mRNA used in this study.

Supplementary Data 2 (download XLSX )

Uncropped gels of Supplementary Figs. 15 and 26c.

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Ren, Q., Zhao, X., Zhou, L. et al. Polymer–mRNA complexes for monocyte-trafficked, lymph node-targeted cancer vaccination. Nat. Biomed. Eng (2026). https://doi.org/10.1038/s41551-026-01672-0

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