Abstract
The blood–brain barrier (BBB) is the main obstacle to the effective delivery of therapeutic agents to the brain, compromising treatment efficacy for a variety of neurological disorders. Intra-arterial (IA) injection of hyperosmotic mannitol has been used to permeabilize the BBB and improve parenchymal entry of therapeutic agents following IA delivery in preclinical and clinical studies. However, the reproducibility of IA BBB manipulation is low and therapeutic outcomes are variable. We demonstrated that this variability could be highly reduced or eliminated when the procedure of osmotic BBB opening is performed under the guidance of interventional MRI. Studies have reported the utility and applicability of this technique in several species. Here we describe a protocol to open the BBB by IA injection of hyperosmotic mannitol under the guidance of MRI in mice. The procedures (from preoperative preparation to postoperative care) can be completed within ~1.5 h, and the skill level required is on par with the induction of middle cerebral artery occlusion in small animals. This MRI-guided BBB opening technique in mice can be utilized to study the biology of the BBB and improve the delivery of various therapeutic agents to the brain.
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Data availability
Source data are provided with this paper. The other data that support the findings of this study are available from the corresponding author upon reasonable request.
Code availability
The code used in this study is provided in Supplementary Data 1. We have also deposited the code and a demonstration of image processing at https://github.com/dychuchengyan/ChengyanMRI.
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Acknowledgements
This work was financially supported by 2017-MSCRFF-3942, 2019-MSCRFF-5031, NIH R01NS091110, R01NS102675 and R21NS091599. We thank I.-H. Wu for preparing Fig. 1 and B. Pocta for editorial assistance.
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Contributions
P.W., M.J., M.P., T.M., S.L. and C.C. contributed to conception and design; C.C., A.J., W.J., Y.G. and X.L. conducted the experiments; C.C., Y.G., G.L. and Y.L. analyzed and interpreted the data; C.C. drafted the manuscript, with revision from A.J., Y.G., X.L., Y.L., W.J., G.L., S.L., T.M., M.P., M.J. and P.W.
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M.P., M.J. and P.W. are founders and equity holders in Intra-ART. M.J. and P.W. are founders and equity holders in Ti-Com.
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Peer review information Nature Protocols thanks Mark S. Bolding, Laura M. Vecchio and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
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Key references using this protocol
Chu, C. et al. Front. Neurol. 9, 921 (2018): https://doi.org/10.3389/fneur.2018.00921
Lesniak, W. G. et al. J. Nucl. Med. 60, 617–622 (2019): https://doi.org/10.2967/jnumed.118.218792
Janowski, M. et al. J. Cereb. Blood Flow Metab. 36, 569–575 (2016): https://doi.org/10.1177/0271678X15615875
Supplementary information
Supplementary Information
Supplementary Figs. 1–4 and Supplementary Methods.
Supplementary Data 1
Matlab code and a demonstration of image processing
Supplementary Video 1
Temporary ligation of ECA and OA cauterization
Supplementary Video 2
Temporary ligation of PPA
Supplementary Video 3
Catheter cannulation
Supplementary Video 4
Brain perfusion of a contrast agent under real-time MRI
Supplementary Video 5
Postoperative procedures
Source data
Source Data Fig. 8
Statistical source data.
Source Data Fig. 9
Statistical source data.
Source Data Fig. 10
Statistical source data.
Source Data Fig. 11
Statistical source data.
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Chu, C., Jablonska, A., Gao, Y. et al. Hyperosmolar blood–brain barrier opening using intra-arterial injection of hyperosmotic mannitol in mice under real-time MRI guidance. Nat Protoc 17, 76–94 (2022). https://doi.org/10.1038/s41596-021-00634-x
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DOI: https://doi.org/10.1038/s41596-021-00634-x
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