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In situ programming of leukaemia-specific T cells using synthetic DNA nanocarriers

Nature Nanotechnology volume 12, pages 813–820 (2017)Cite this article

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Abstract

An emerging approach for treating cancer involves programming patient-derived T cells with genes encoding disease-specific chimeric antigen receptors (CARs), so that they can combat tumour cells once they are reinfused. Although trials of this therapy have produced impressive results, the in vitro methods they require to generate large numbers of tumour-specific T cells are too elaborate for widespread application to treat cancer patients. Here, we describe a method to quickly program circulating T cells with tumour-recognizing capabilities, thus avoiding these complications. Specifically, we demonstrate that DNA-carrying nanoparticles can efficiently introduce leukaemia-targeting CAR genes into T-cell nuclei, thereby bringing about long-term disease remission. These polymer nanoparticles are easy to manufacture in a stable form, which simplifies storage and reduces cost. Our technology may therefore provide a practical, broadly applicable treatment that can generate anti-tumour immunity ‘on demand’ for oncologists in a variety of settings.

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Figure 1: Design and manufacture of lymphocyte-programming nanoparticles.
Figure 2: DNA nanocarriers choreograph robust and persistent CAR production by lymphocytes in vitro.
Figure 3: CD3-targeted nanoparticles bind to circulating T cells in mice.
Figure 4: Reprogramming host T cells with leukaemia-specific CAR genes.
Figure 5: Nanoparticle-programmed CAR lymphocytes can cause tumour regression with efficacies similar to adoptive T-cell therapy.

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Acknowledgements

We thank I. Stanishevskaya and D. Ehlert (cognitionstudio.com) for the design of the illustrations. We also thank M. Sadelain (Memorial Sloan-Kettering Cancer Center, New York, New York) for the Eμ-ALL01 cell line, and for the DNA construct that encodes an all-murine CD19-specific CAR. This work was supported in part by the Fred Hutchinson Cancer Research Center's Immunotherapy Initiative with funds provided by the Bezos Family Foundation, a New Idea Award from the Leukemia & Lymphoma Society, the Phi Beta Psi Sorority, the National Science Foundation (CAREER, award no. 1452492 and EAGER award no. 1644363), and the National Cancer Institute of the National Institutes of Health under award no. R01CA207407. M.T.S. was also supported by a Research Scholar Grant (RSG-16-110-01 – LIB) from the American Cancer Society.

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  1. Tyrel T. Smith, Sirkka B. Stephan and Howell F. Moffett: These authors contributed equally to this work.

Authors and Affiliations

  1. Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, 98109, Washington, USA

    Tyrel T. Smith, Sirkka B. Stephan, Howell F. Moffett, Laura E. McKnight, Weihang Ji, Martin E. Wohlfahrt & Matthias T. Stephan

  2. Technology Access Foundation (TAF) Academy, Fred Hutchinson Cancer Research Center, Seattle, 98109, Washington, USA

    Diana Reiman, Emmy Bonagofski & Matthias T. Stephan

  3. Comparative Pathology, Fred Hutchinson Cancer Research Center, Seattle, 98109, Washington, USA

    Smitha P. S. Pillai

  4. Department of Bioengineering and Molecular Engineering & Sciences Institute, University of Washington, Seattle, 98105, Washington, USA

    Matthias T. Stephan

  5. Department of Medicine, Division of Medical Oncology, University of Washington, Seattle, 98109, Washington, USA

    Matthias T. Stephan

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Contributions

T.T.S., S.B.S. and H.F.M. designed and performed experiments and analysed and interpreted data. L.E.M. helped clone plasmid vectors, W.J. synthesized the PBAE polymer, and D.R. and E.B. helped with the large-scale purification of CAR-encoding plasmid DNA. M.E.W. performed the Southern blot analysis. S.P.S.P. performed and analysed in vivo safety/toxicity studies, and M.T.S. designed the study, performed experiments, analysed and interpreted data, and wrote the manuscript.

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Correspondence to Matthias T. Stephan.

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The authors declare no competing financial interests.

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Smith, T., Stephan, S., Moffett, H. et al. In situ programming of leukaemia-specific T cells using synthetic DNA nanocarriers. Nature Nanotech 12, 813–820 (2017). https://doi.org/10.1038/nnano.2017.57

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