Abstract

The neurovascular unit (NVU) regulates metabolic homeostasis as well as drug pharmacokinetics and pharmacodynamics in the central nervous system. Metabolic fluxes and conversions over the NVU rely on interactions between brain microvascular endothelium, perivascular pericytes, astrocytes and neurons, making it difficult to identify the contributions of each cell type. Here we model the human NVU using microfluidic organ chips, allowing analysis of the roles of individual cell types in NVU functions. Three coupled chips model influx across the blood–brain barrier (BBB), the brain parenchymal compartment and efflux across the BBB. We used this linked system to mimic the effect of intravascular administration of the psychoactive drug methamphetamine and to identify previously unknown metabolic coupling between the BBB and neurons. Thus, the NVU system offers an in vitro approach for probing transport, efficacy, mechanism of action and toxicity of neuroactive drugs.

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Acknowledgements

This research was supported by the Wyss Institute for Biologically Inspired Engineering at Harvard University, Defense Advanced Research Projects Agency (DARPA) under Cooperative Agreement Number W911NF-12-2-0036 (D.E.I. & K.K.P.), and Sverige-Amerika Stiftelsen, Carl Trygger Stiftelse, Erik och Edith Fernstrom's stiftelse (A.H.). The views and conclusions contained in this document are those of the authors and should not be interpreted as representing the official policies, either expressed or implied, of DARPA or the US Government. This work also was made possible by access to the microfabrication facilities of the Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Coordinated Infrastructure Network (NNCI), which is supported by the National Science Foundation under NSF award no. 1541959. CNS is part of Harvard University. We also thank T. Ferrante for technical assistance, M. Rosnach and J.P. Ferrier for artwork and technical illustrations, to J.A. Goss for his help with chips design and fabrication, and the Harvard Medical School Neurobiology Imaging Facility (supported in part by NINDS P30 Core Center grant #NS072030) for consultation and instrument availability.

Author information

Author notes

    • Ben M Maoz
    • , Anna Herland
    •  & Edward A FitzGerald

    These authors contributed equally to this work.

Affiliations

  1. Disease Biophysics Group, Harvard John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA.

    • Ben M Maoz
    • , Thomas Grevesse
    • , Sean P Sheehy
    • , Stephanie Dauth
    • , Nikita Budnik
    • , Kevin Shores
    • , Alexander Cho
    • , Janna C Nawroth
    •  & Kevin Kit Parker
  2. Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts, USA.

    • Ben M Maoz
    • , Anna Herland
    • , Edward A FitzGerald
    • , Thomas Grevesse
    • , Alan R Pacheco
    • , Sean P Sheehy
    • , Tae-Eun Park
    • , Stephanie Dauth
    • , Robert Mannix
    • , Kevin Shores
    • , Alexander Cho
    • , Janna C Nawroth
    • , Donald E Ingber
    •  & Kevin Kit Parker
  3. Department of Biomedical Engineering, Faculty of Engineering, Tel Aviv University, Tel Aviv, Israel.

    • Ben M Maoz
  4. Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel.

    • Ben M Maoz
  5. The Center for Nanoscience and Nanotechnology, Tel Aviv University, Tel Aviv, Israel.

    • Ben M Maoz
  6. Department of Micro and Nanosystems, KTH Royal Institute of Technology, Stockholm, Sweden.

    • Anna Herland
  7. Swedish Medical Nanoscience Center, Department of Neuroscience, Karolinska Institute, Stockholm, Sweden.

    • Anna Herland
  8. Small Molecule Mass Spectrometry Facility, Harvard University, Cambridge, Massachusetts, USA.

    • Charles Vidoudez
  9. Graduate Program in Bioinformatics and Biological Design Center, Boston University, Boston, Massachusetts, USA.

    • Alan R Pacheco
    •  & Daniel Segrè
  10. Vascular Biology Program and Department of Surgery, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts, USA.

    • Robert Mannix
    •  & Donald E Ingber
  11. Department of Biology, Department of Biomedical Engineering, Department of Physics, Boston University, Boston, Massachusetts, USA.

    • Daniel Segrè
  12. Mass Spectrometry and Proteomics Resource Laboratory, Harvard University, Cambridge, Massachusetts, USA.

    • Bogdan Budnik
  13. Harvard John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA.

    • Donald E Ingber

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Contributions

B.M.M., A.H., E.A.F., T.G., D.E.I. and K.K.P. designed the study. C.V. ran and analyzed the MS samples, and A.R.P. and D.S. performed the flux balance analysis modeling. S.P.S. performed bioinformatic analysis for proteomics and MS. A.H., E.A.F. and T.-E.P. conducted the BBB chip culture and B.M.M., T.G. and s.d. performed brain chip culture and imaging. B.M.M., A.H., E.A.F., T.G., S.D. and R.M. performed confocal imaging, N.B. and B.B. conducted proteomic run and analysis. K.S. performed the COMSOL modeling, B.M.M., T.G. and A.C. conducted sample preparation for brain chip. B.M.M., T.G. and J.C.N. contributed to the brain chip design. B.M.M., A.H., E.A.F., T.G., D.E.I. and K.K.P. prepared illustrations and wrote the manuscript.

Competing interests

D.E.I. holds equity in Emulate, Inc., consults for the company, and chairs its scientific advisory board. K.K.P. is a consultant and a member of the Scientific Advisory Board of Emulate, Inc.

Corresponding authors

Correspondence to Donald E Ingber or Kevin Kit Parker.

Integrated supplementary information

Supplementary information

PDF files

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    Supplementary Text and Figures

    Supplementary Figures 1–12

  2. 2.

    Life Sciences Reporting Summary

  3. 3.

    Supplementary Information

    Supplementary Tables 1–11 and Supplementary Scripts 1–5

Videos

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    Video 1

    Hypoxia conditions in the chips

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    Video 2

    Cascade blue diffusion in the system

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    Video 3

    Proteomaps of uncoupled BBB Chip endo

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    Video 4

    Proteomaps of coupled BBB Chip endo

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    Video 5

    Proteomaps of uncoupled BBB Chipastro/peri

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    Video 6

    Proteomaps of coupled BBB Chip astro/peri

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    Video 7

    Proteomaps of uncoupled Brain Chip

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    Video 8

    Proteomaps of coupled Brain Chip

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    Video 9

    Live imaging of the Brain Chip

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DOI

https://doi.org/10.1038/nbt.4226