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Photothermal nanofibres enable safe engineering of therapeutic cells

Nature Nanotechnology volume 16, pages 1281–1291 (2021)Cite this article

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Abstract

Nanoparticle-sensitized photoporation is an upcoming approach for the intracellular delivery of biologics, combining high efficiency and throughput with excellent cell viability. However, as it relies on close contact between nanoparticles and cells, its translation towards clinical applications is hampered by safety and regulatory concerns. Here we show that light-sensitive iron oxide nanoparticles embedded in biocompatible electrospun nanofibres induce membrane permeabilization by photothermal effects without direct cellular contact with the nanoparticles. The photothermal nanofibres have been successfully used to deliver effector molecules, including CRISPR–Cas9 ribonucleoprotein complexes and short interfering RNA, to adherent and suspension cells, including embryonic stem cells and hard-to-transfect T cells, without affecting cell proliferation or phenotype. In vivo experiments furthermore demonstrated successful tumour regression in mice treated with chimeric antibody receptor T cells in which the expression of programmed cell death protein 1 (PD1) is downregulated after nanofibre photoporation with short interfering RNA to PD1. In conclusion, cell membrane permeabilization with photothermal nanofibres is a promising concept towards the safe and more efficient production of engineered cells for therapeutic applications, including stem cell or adoptive T cell therapy.

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Fig. 1: Concept of intracellular delivery by photothermal nanofibres and characterization of photothermal electrospun nanofibres.
Fig. 2: PEN photoporation enables safe and efficient delivery of macromolecules to cells.
Fig. 3: PEN photoporation for siRNA gene silencing or CRISPR–Cas9-mediated gene knockout in H1299.
Fig. 4: PEN photoporation enables efficient intracellular delivery of macromolecules into hESCs.
Fig. 5: PEN photoporation enables efficient intracellular delivery of siRNA into T cells.
Fig. 6: PEN photoporation also retains T cell effector functions in vivo.

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Data availability

All data supporting the findings of this study are available within the paper and its Supplementary Information. Source data are provided with this paper. Any further related information can be provided by the corresponding author upon reasonable request.

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (grant nos 21774060 and 21644004), the European Research Council (ERC Consolidator Grant 648124), the Research Foundation Flanders (FWO, 1500418N, 12Q8718N and 1S62517N), the National Key R&D Program of China (2017YFF0207804), and the Youth Innovation Promotion Association CAS (grant no. 2018491). We acknowledge funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement number 810685 (DelNam project).

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Authors and Affiliations

  1. Joint Laboratory of Advanced Biomedical Materials (Nanjing Forestry University–Ghent University), International Innovation for Center for Forest Chemicals and Materials, College of Chemical Engineering, Nanjing Forestry University, Nanjing, People’s Republic of China

    Ranhua Xiong, Dawei Hua, Chaobo Huang & Stefaan C. De Smedt

  2. Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium

    Ranhua Xiong, Dawei Hua, Jelter Van Hoeck, Dominika Berdecka, Juan C. Fraire, Laurens Raes, Aranit Harizaj, Félix Sauvage, Koen Raemdonck, Stefaan C. De Smedt & Kevin Braeckmans

  3. Laboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp, Antwerp, Belgium

    Dominika Berdecka & Winnok H. De Vos

  4. Department of Human Structure and Repair, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium

    Laurens Léger, Jeffrey Aalders & Jolanda van Hengel

  5. Department of Diagnostic Sciences and Cancer Research Institute Ghent, Ghent University, Ghent, Belgium

    Stijn De Munter

  6. Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium

    Glenn Goetgeluk, Melissa Pille & Bart Vandekerckhove

  7. Department of Chemistry, Atomic and Mass Spectrometry Research Group, Ghent University, Ghent, Belgium

    Joke Belza, Thibaut Van Acker, Eduardo Bolea-Fernandez & Frank Vanhaecke

  8. Department of Modern Mechanics, University of Science and Technology of China, Hefei, People’s Republic of China

    Ting Si

  9. Center for Advanced Light Microscopy, Ghent University, Ghent, Belgium

    Stefaan C. De Smedt & Kevin Braeckmans

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  1. Ranhua Xiong
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Contributions

R.X., K.B. and S.C.D.S. conceived the concept of PEN photoporation for contact-free photoporation and designed the experiments. R.X., C.H. and D.H. fabricated and characterized the PEN substrates. R.X. performed the PEN photoporation experiments and analysed the data, and also performed the CFD simulations together with J.C.F. J.B., E.B.-F. and T.V.A. performed and analysed the ICP-MS/MS measurements. L.R. and M.P. prepared the RNPs. L.L. and J.A. were involved in the preparation and PEN photoporation of hESCs. J.V.H., D.B., S.D.M., K.R., G.G., A.H., F.S. and B.V. were involved in the preparation and PEN photoporation of human T cells. K.B., S.C.D.S., C.H., K.R., T.S., F.V., J.V.H., W.H.D.V. and B.V. advised on experiments and data analysis. All authors discussed the experimental results and jointly wrote the manuscript.

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Correspondence to Ranhua Xiong, Chaobo Huang, Stefaan C. De Smedt or Kevin Braeckmans.

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The authors declare no competing interests.

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Peer review information Nature Nanotechnology thanks Massimiliano Caiazzo and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Tables 1–3, Notes 1–3 and Figs. 1–19.

Reporting Summary

Supplementary Video 1

A 3D volume rendering of photothermal electrospun nanofibres.

Supplementary Video 2

Numerical simulation of the heat transfer from a single IONP to the surrounding medium via the surrounding nanofibre. Black solid circles in the top right and bottom right indicate the profile of nanoparticle. The time unit in the bottom left is second. The orange dashed lines indicate the boundary of the nanofibre. The simulation conditions were h = 40 nm, N = 1 and I = 0.08 J cm–2.

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Xiong, R., Hua, D., Van Hoeck, J. et al. Photothermal nanofibres enable safe engineering of therapeutic cells. Nat. Nanotechnol. 16, 1281–1291 (2021). https://doi.org/10.1038/s41565-021-00976-3

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