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Electroporation-mediated novel albumin-fused Flt3L DNA delivery promotes cDC1-associated anticancer immunity

Gene Therapy volume 32, pages 277–286 (2025)Cite this article

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Abstract

Dendritic cells (DCs) constitute a distinct type of immune cell found within tumors, serving a central role in mediating tumor antigen-specific immunity against cancer cells. Frequently, DC functions are dysregulated by the immunosuppressive signals present within the tumor microenvironment (TME). Consequently, DC manipulation holds great potential to enhance the cytotoxic T cell response against cancer diseases. One strategy involves administering Fms-like tyrosine kinase receptor 3 ligand (Flt3L), a vitally important cytokine for DC development. In this current study, the electroporation-mediated delivery of a novel albumin-fused Flt3L DNA (alb-Flt3L DNA) demonstrated the ability to induce an anti-tumor immune response. This albumin fusion construct possesses more persistent bioactivity in targeted organs. Furthermore, TC-1-bearing-C57BL/6 mice receiving alb-Flt3L DNA treatment presented better tumor control and superior survival. Cellular analysis revealed that alb-Flt3L DNA administration promoted robust DC and cDC1 expansion. In addition, increased levels of IFN-γ-secreting CD8+ lymphocytes were found in correlation to greater cDC1 population. Moreover, the toxicity of alb-Flt3L administration is limited. Collectively, our data showcases a novel DC-based immunotherapy using electroporation to administer alb-Flt3L DNA.

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Fig. 1: Albumin-fused Gaussia luciferase (alb-GLuc) exhibits extended biodistribution under local injection compared to GLuc.
Fig. 2: alb-Flt3L DNA treatment elicits superior tumor control and better survival compared with that of Flt3L DNA.
Fig. 3: alb-Flt3L DNA-induced DC expansion leads to tumor control.
Fig. 4: alb-Flt3L DNA demonstrates robust anti-tumor immunity against TC-1 tumor.
Fig. 5: Histology examination of key organs of mice receiving treatments.

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Data availability

Raw data were generated at Johns Hopkins University School of Medicine. The datasets used and/or analyzed during the current study available from the corresponding author on reasonable request.

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Acknowledgements

This study was supported by the National Institute of Health, National Cancer Institute Specialized Program of Research Excellence (SPORE) in Cervical Cancer grant (NIH/NCI P50CA098252) and NCI awards (R01CA237067, R21DE029910-01, R21CA256020, and 1R21CA234516-01A1).

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Authors and Affiliations

  1. Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Ming-Hung Hu, Darrell Fan, Hsin-Fang Tu, Ya-Chea Tsai, Liangmei He, Zhicheng Zhou, Michelle Cheng, Deyin Xing, Suyang Wang, Alexis Wu, T. C. Wu & Chien-Fu Hung

  2. Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan, ROC

    Ming-Hung Hu

  3. Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan, ROC

    Ming-Hung Hu

  4. Cancer Center, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan, ROC

    Ming-Hung Hu

  5. Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    T. C. Wu & Chien-Fu Hung

  6. Department of Obstetrics and Gynecology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    T. C. Wu

  7. Molecular Microbiology and Immunology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    T. C. Wu

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Contributions

Conception and design: M.H.H. and C.F.H. Conducting experiments: M.H.H., D.F., Y.C.T., L.H., and Z.Z. analysis and interpretation of data: M.H.H., D.F., H.F.T., D.X., and S.W. Writing and review of manuscript: M.H.H., D.F., M.C., A.W., and C.F.H. Study supervision: T.C.W. and C.F.H.

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Correspondence to Chien-Fu Hung.

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All mouse experiments were conducted at the Johns Hopkins University School of Medicine following preapproved protocols and in accordance with the Johns Hopkins University School of Medicine Institutional Animal Care and Use Committee (MO23M84).

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Hu, MH., Fan, D., Tu, HF. et al. Electroporation-mediated novel albumin-fused Flt3L DNA delivery promotes cDC1-associated anticancer immunity. Gene Ther 32, 277–286 (2025). https://doi.org/10.1038/s41434-024-00497-3

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