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Efficacy and safety assessment of a TRAF6-targeted nanoimmunotherapy in atherosclerotic mice and non-human primates

Nature Biomedical Engineering volume 2, pages 279–292 (2018)Cite this article

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Abstract

Macrophage accumulation in atherosclerosis is directly linked to the destabilization and rupture of plaque, causing acute atherothrombotic events. Circulating monocytes enter the plaque and differentiate into macrophages, where they are activated by CD4+ T lymphocytes through CD40–CD40 ligand signalling. Here, we report the development and multiparametric evaluation of a nanoimmunotherapy that moderates CD40–CD40 ligand signalling in monocytes and macrophages by blocking the interaction between CD40 and tumour necrosis factor receptor-associated factor 6 (TRAF6). We evaluated the biodistribution characteristics of the nanoimmunotherapy in apolipoprotein E-deficient (Apoe–/–) mice and in non-human primates by in vivo positron-emission tomography imaging. In Apoe–/– mice, a 1-week nanoimmunotherapy treatment regimen achieved significant anti-inflammatory effects, which was due to the impaired migration capacity of monocytes, as established by a transcriptome analysis. The rapid reduction of plaque inflammation by the TRAF6-targeted nanoimmunotherapy and its favourable toxicity profiles in both mice and non-human primates highlights the translational potential of this strategy for the treatment of atherosclerosis.

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Fig. 1: TRAF6i–HDL biodistribution and uptake.
Fig. 2: TRAF6i–HDL biodistribution in non-human primates.
Fig. 3: TRAF6i–HDL therapy decreases plaque macrophage content as assessed by histology.
Fig. 4: TRAF6i–HDL decreases plaque inflammation due to impaired Ly6Chi monocyte recruitment.
Fig. 5: TRAF6i–HDL shows effects on monocyte migration, among other affected processes.
Fig. 6: TRAF6i–HDL therapy does not elicit adverse immune or toxic effects in mice and non-human primates.

Change history

  • 26 July 2018

    In the version of this Article originally published, the surname of the author Edward A. Fisher was spelt incorrectly as ‘Fischer’. This has now been corrected.

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Acknowledgements

The authors thank the Icahn School of Medicine and the following Mount Sinai’s core facilities: flow cytometry core, quantitative PCR core and TMII’s preclinical imaging core. This study was funded by National Institutes of Health grants R01 HL118440, R01 HL125703 and P01 HL131478 (all to W.J.M.M.), R01 EB009638 (to Z.A.F.) and R01 HL144072 (to W.J.M.M. and Z.A.F.), as well as by NWO grants ZonMW Veni 016156059 (to R.D.) and ZonMW Vidi 91713324 (to W.J.M.M.), and by the European Research Council (ERC Con to E.L.) and by the DFG (SFB 1123-A5 to E.L.).

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Authors and Affiliations

  1. Translational and Molecular Imaging Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    Marnix Lameijer, Mandy M. T. van Leent, Max L. Senders, Francois Fay, Joost Malkus, Brenda L. Sanchez-Gaytan, Abraham J. P. Teunissen, Nicolas Karakatsanis, Philip Robson, Carlos Pérez-Medina, Claudia Calcagno, Zahi A. Fayad, Willem J. M. Mulder & Raphaël Duivenvoorden

  2. Department of Medical Biochemistry, Academic Medical Center, Amsterdam, The Netherlands

    Marnix Lameijer, Mandy M. T. van Leent, Max L. Senders, Tom T. P. Seijkens, Esther Lutgens & Willem J. M. Mulder

  3. Cluster for Molecular Imaging and Department of Clinical Physiology, Nuclear Medicine and PET, Rigshospitalet and University of Copenhagen, Copenhagen, Denmark

    Tina Binderup & Andreas Kjaer

  4. Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    Xianxiao Zhou & Bin Zhang

  5. Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Yuxiang Ye, Gregory Wojtkiewicz, Filip K. Swirski & Matthias Nahrendorf

  6. Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, NY, USA

    Jun Tang & Thomas Reiner

  7. Department of Vascular Medicine, Amsterdam Cardiovascular Sciences, Academic Medical Center, Amsterdam, The Netherlands

    Jeffrey Kroon, Erik S. G. Stroes & Raphaël Duivenvoorden

  8. Immunology Institute, Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    Jordi Ochando

  9. Department of Medicine (Cardiology) and Cell Biology, Marc and Ruti Bell Program in Vascular Biology, NYU School of Medicine, New York, NY, USA

    Edward A. Fisher

  10. Saha Cardiovascular Research Center and Department of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, USA

    Ryan E. Temel

  11. Institute for Cardiovascular Prevention, Ludwig-Maximilians University, Munich, Germany

    Esther Lutgens

  12. Department of Nephrology, Academic Medical Center, Amsterdam, The Netherlands

    Raphaël Duivenvoorden

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  1. Marnix Lameijer
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Contributions

R.D. and W.J.M.M. designed the study. R.D., M.L., T.B., M.M.T.v.L., M.L.S., J.T., T.T.P.S., J.K., E.S.G.S., J.O., E.A.F., R.E.T., N.K., P.R., A.K., F.K.S., M.N., Z.A.F., E.L. and W.J.M.M. designed, performed and oversaw the in vivo and ex vivo experiments. F.F., B.L.S.-G. and M.L. developed and produced TRAF6i–HDL. Flow cytometry, histology and immunostaining, laser capture microdissection, and blood chemistry experiments were performed and analysed by R.D., M.L., M.M.T.v.L. and J.M. FMT/CT was performed and analysed by R.D., Y.Y., G.W. and M.N. The RNA sequencing was performed and analysed by X.Z., B.Z., R.D. and M.L. Monocyte migration assays were performed by J.K. PET/CT and pharmacokinetic studies in mice were performed by C.P.-M., J.T. and T.R. 89Zr-PET/MRI in non-human primates was performed and analysed by T.B., M.L.S., C.P.-M. and C.C. The manuscript was written by R.D., M.L. and W.J.M.M. All authors contributed to the writing of the manuscript and approved the final draft. R.D., Z.A.F. and W.J.M.M. provided funding.

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Correspondence to Willem J. M. Mulder or Raphaël Duivenvoorden.

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Supplementary information

Supplementary Information

Supplementary figures, tables and video captions.

Reporting Summary

Supplementary Video 1

Three-dimensional MRI of a non-human primate.

Supplementary Video 2

Three-dimensional PET distribution of 89Zr-labelled TRAF6i–HDL at 60 minutes in a non-human primate.

Supplementary Video 3

In vitro transendothelial migration of monocytes pre-treated with TRAF6i–HD.

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Lameijer, M., Binderup, T., van Leent, M.M.T. et al. Efficacy and safety assessment of a TRAF6-targeted nanoimmunotherapy in atherosclerotic mice and non-human primates. Nat Biomed Eng 2, 279–292 (2018). https://doi.org/10.1038/s41551-018-0221-2

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