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A pH-responsive nanoparticle targets the neurokinin 1 receptor in endosomes to prevent chronic pain

Nature Nanotechnology volume 14, pages 1150–1159 (2019)Cite this article

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Abstract

Nanoparticle-mediated drug delivery is especially useful for targets within endosomes because of the endosomal transport mechanisms of many nanomedicines within cells. Here, we report the design of a pH-responsive, soft polymeric nanoparticle for the targeting of acidified endosomes to precisely inhibit endosomal signalling events leading to chronic pain. In chronic pain, the substance P (SP) neurokinin 1 receptor (NK1R) redistributes from the plasma membrane to acidified endosomes, where it signals to maintain pain. Therefore, the NK1R in endosomes provides an important target for pain relief. The pH-responsive nanoparticles enter cells by clathrin- and dynamin-dependent endocytosis and accumulate in NK1R-containing endosomes. Following intrathecal injection into rodents, the nanoparticles, containing the FDA-approved NK1R antagonist aprepitant, inhibit SP-induced activation of spinal neurons and thus prevent pain transmission. Treatment with the nanoparticles leads to complete and persistent relief from nociceptive, inflammatory and neuropathic nociception and offers a much-needed non-opioid treatment option for chronic pain.

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Fig. 1: Characterization of DIPMA and BMA nanoparticles.
Fig. 2: Uptake and disassembly of DIPMA and BMA nanoparticles in HEK-293 cells.
Fig. 3: Biodistribution and cellular uptake of nanoparticles and aprepitant delivery.
Fig. 4: Effects of nanoparticles on nociceptive, inflammatory and neuropathic nociception.
Fig. 5: Sensitization and activation of nociceptive transmission.
Fig. 6: Antagonism of NK1R signalling in endosomes.

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Data availability

All data generated or analysed during this study are available in this Article and its Supplementary Information or from the corresponding authors upon request.

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Acknowledgements

This work was supported by the National Institutes of Health (NS102722, DE026806, DK118971), the Department of Defense (PR170507), the National Health and Medical Research Council (63303, 1049682, 1031886; N.W.B.), the Australian Research Council Centre of Excellence in Convergent Bio-Nano Science and Technology (N.W.B. and T.P.D.), the Center for the Development of Nanoscience and Nanotechnology (CEDENNA, Fondecyt no. 1181622, L.C.) and Takeda Pharmaceuticals Inc. (N.W.B., N.A.V and D.P.P.). We thank F. Chiu for mass spectrometry analysis of aprepitant loading and P. Zhao for advice about signalling assays.

Author information

Authors and Affiliations

  1. Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia

    Paulina D. Ramírez-García, Jeffri S. Retamal, Priyank Shenoy, Nghia Truong, Cameron J. Nowell, Song Y. Khor, Louis M. Layani, Chris Lumb, Daniel P. Poole, TinaMarie Lieu, Gregory D. Stewart, Quynh N. Mai, John F. Quinn, Michael R. Whittaker, Nicholas A. Veldhuis, Thomas P. Davis & Nigel W. Bunnett

  2. Australian Research Council Centre of Excellence in Convergent Bio-Nano Science and Technology, Monash University, Parkville, Victoria, Australia

    Paulina D. Ramírez-García, Jeffri S. Retamal, Priyank Shenoy, Nghia Truong, Song Y. Khor, Louis M. Layani, Chris Lumb, Daniel P. Poole, TinaMarie Lieu, Quynh N. Mai, John F. Quinn, Michael R. Whittaker, Nicholas A. Veldhuis, Thomas P. Davis & Nigel W. Bunnett

  3. Monash Biomedicine Discovery Institute, Monash University, Melbourne, Victoria, Australia

    Wendy Imlach & Matthew Sykes

  4. Laboratory of Neurobiology, Center for the Development of Nanoscience and Nanotechnology (CEDENNA), University of Santiago de Chile, Santiago, Chile

    Luis Constandil & Teresa Pelissier

  5. Departments of Surgery and Pharmacology, Columbia University Vagelos College of Physicians and Surgeons, Columbia University in the City of New York, New York , NY, USA

    Dane D. Jensen, Rocco Latorre & Nigel W. Bunnett

  6. Bluestone Centre for Clinical Research, New York University College of Dentistry, New York, NY, USA

    Nicole N. Scheff & Brian L. Schmidt

  7. Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, Queensland, Australia

    Thomas P. Davis

  8. Department of Pharmacology and Therapeutics, University of Melbourne, Parkville, Victoria, Australia

    Nigel W. Bunnett

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  1. Paulina D. Ramírez-García
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Contributions

P.D.R.-G. prepared and characterized nanoparticles, examined nanoparticle uptake and disassembly, studied SP signalling in model cells and wrote the manuscript. J.S.R. studied the biodistribution and anti-nociceptive and in vivo electrophysiological actions of nanoparticles. P.S. studied the biodistribution and anti-nociceptive actions of nanoparticles. W.I. conceived and designed electrophysiological studies on spinal neurons. M.S. studied the excitation of spinal neurons, and N.T. prepared and characterized nanoparticles. L.C. conceived and designed neuropathic nociception and in vivo electrophysiological studies. T.P. conceived and designed neuropathic nociception. C.J.N. provided expertise in the analysis of confocal images and S.Y.K. obtained transmission electron microscopy images. L.M.L. characterized the critical micellar concentration and pH-disassembly of nanoparticles. C.L. studied SP signalling in model cells and D.P.P. studied nanoparticle uptake. T.M.L. studied anti-nociceptive actions of nanoparticles, G.D.S. prepared striatal neurons, and Q.N.M. prepared and characterized nanoparticles. D.D.J. examined NK1R endocytosis, nanoparticle uptake into spinal neurons, and SP signalling in model cells and striatal neurons. R.L. examined NK1R endocytosis and nanoparticle uptake into spinal neurons. N.S.N. studied NK1R endocytosis in rats. B.L.S. designed experiments to examine NK1R endocytosis in rats. J.F.Q. designed nanoparticles and wrote the manuscript. M.R.W. designed nanoparticles. N.A.V. conceived experiments, studied SP signalling in neurons, interpreted the results and wrote the manuscript. T.P.D. conceived the experiments and designed the nanoparticles. N.W.B. conceived and designed the experiments, interpreted the results and wrote the manuscript.

Corresponding authors

Correspondence to Nicholas A. Veldhuis, Thomas P. Davis or Nigel W. Bunnett.

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Competing interests

Research in N.A.V.'s, D.P.P.’s and N.W.B.’s laboratories is funded, in part, by Takeda Pharmaceuticals. N.W.B. is a founding scientist of Endosome Therapeutics.

Additional information

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

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary information

Supplementary Methods, Supplementary Figs. 1–8, Supplementary Video titles and legends 1–7 and Supplementary refs. 1–4.

Reporting Summary

Supplementary Video 1

Localization of DIPMA-Cy5 nanoparticles and Rab5a-GFP in HEK-293 cells.

Supplementary Video 2

Localization of DIPMA-Cy5 nanoparticles and Rab7a-GFP in HEK-293 cells.

Supplementary Video 3

Localization of DIPMA-Cy5 nanoparticles and NK1R-GFP in HEK-293 cells.

Supplementary Video 4

Localization of DIPMA-Cy5 nanoparticles in the mouse dorsal horn.

Supplementary Video 5

Localization of BMA-Cy5 nanoparticles in the mouse dorsal horn.

Supplementary Video 6

Localization of NK1R-IR in the rat dorsal horn after sham surgery.

Supplementary Video 7

Localization of NK1R-IR in the rat dorsal horn after SNS surgery.

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Ramírez-García, P.D., Retamal, J.S., Shenoy, P. et al. A pH-responsive nanoparticle targets the neurokinin 1 receptor in endosomes to prevent chronic pain. Nat. Nanotechnol. 14, 1150–1159 (2019). https://doi.org/10.1038/s41565-019-0568-x

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