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In vivo compression and imaging in mouse brain to measure the effects of solid stress

Nature Protocols volume 15, pages 2321–2340 (2020)Cite this article

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Abstract

We recently developed an in vivo compression device that simulates the solid mechanical forces exerted by a growing tumor on the surrounding brain tissue and delineates the physical versus biological effects of a tumor. This device, to our knowledge the first of its kind, can recapitulate the compressive forces on the cerebellar cortex from primary (e.g., glioblastoma) and metastatic (e.g., breast cancer) tumors, as well as on the cerebellum from tumors such as medulloblastoma and ependymoma. We adapted standard transparent cranial windows normally used for intravital imaging studies in mice to include a turnable screw for controlled compression (acute or chronic) and decompression of the cerebral cortex. The device enables longitudinal imaging of the compressed brain tissue over several weeks or months as the screw is progressively extended against the brain tissue to recapitulate tumor growth–induced solid stress. The cranial window can be simply installed on the mouse skull according to previously established methods, and the screw mechanism can be readily manufactured in-house. The total time for construction and implantation of the in vivo compressive cranial window is <1 h (per mouse). This technique can also be used to study a variety of other diseases or disorders that present with abnormal solid masses in the brain, including cysts and benign growths.

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Fig. 1: Tumor progression and brain loss are consequences of both physical and biological interactions at the tumor–brain interface.
Fig. 2: The in vivo compressive cranial window is capable of decoupling mechanical stresses from biological stresses at the tumor–brain interface.
Fig. 3: Compressive cranial window recapitulates the compressed brain tissue caused by brain tumors.
Fig. 4: Surgical procedure for implanting a cCW.
Fig. 5: Timing and experimental design of cCW.
Fig. 6: The cCW allows for intravital imaging.
Fig. 7: Procedure for fabrication of a cCW.
Fig. 8: Chronic compression results in cellular biological responses that can be quantified via histological and molecular techniques.
Fig. 9: Chronic compression results in behavioral responses that can be quantified via different functional tests.

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Data availability

The datasets generated and/or analyzed during the current study are available from the corresponding author upon request.

Code availability

The code associated with the datasets generated and/or analyzed during the current study is available from the corresponding author upon request.

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Acknowledgements

This work was supported in part by the National Cancer Institute (P01-CA080124, R35-CA197743, U01-CA224173, and R01-CA208205 to R.K.J.; F32-CA216944 to H.T.N), the American Association of Cancer Research (19-40-50-DATT to M.D.), the Susan G. Komen Foundation (PDF14201739 to G.S.), and the European Research Council (ERC; 805225 to G.S.). R.K.J.’s research is also supported by grants from the National Foundation for Cancer Research, Harvard Ludwig Center, Jane’s Trust Foundation, and the Bill and Melinda Gates Foundation.

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Author notes

  1. These authors contributed equally: Hadi T. Nia, Meenal Datta, Giorgio Seano.

Authors and Affiliations

  1. Edwin L. Steele Laboratories, Department of Radiation Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Hadi T. Nia, Meenal Datta, Giorgio Seano, William W. Ho, Sylvie Roberge, Peigen Huang, Lance L. Munn & Rakesh K. Jain

  2. Department of Biomedical Engineering, Boston University, Boston, MA, USA

    Hadi T. Nia & Sue Zhang

  3. Tumor Microenvironment Laboratory, Institut Curie Research Center, Paris-Saclay University, PSL Research University, Inserm U1021, CNRS UMR3347, Orsay, France

    Giorgio Seano

  4. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

    William W. Ho

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Contributions

H.T.N., M.D., G.S., W.W.H., S.R., P.H., L.L.M., and R.K.J. designed the study; H.T.N., M.D., G.S., S.R., and P.H. acquired the data. H.T.N., M.D., G.S., and R.K.J. contributed to analysis and interpretation of the data. H.T.N., M.D., G.S., S.Z., W.W.H., S.R., P.H., L.L.M., and R.K.J. were involved in drafting the article and revising it for important intellectual content.

Corresponding author

Correspondence to Rakesh K. Jain.

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Competing interests

R.K.J. received an honorarium from Amgen; consultant fees from Chugai, Enlight, Merck, Ophthotech, Pfizer, SPARC, SynDevRx, and XTuit; owns equity in Enlight, Ophthotech, and SynDevRx; and serves on the boards of trustees of Tekla Healthcare Investors, Tekla Life Sciences Investors, Tekla Healthcare Opportunities Fund, and Tekla World Healthcare Fund. No funding or reagents from these companies were used in this study.

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Seano, G. et al. Nat. Biomed. Eng. 3, 230–245 (2019): https://doi.org/10.1038/s41551-018-0334-7

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Nia, H.T., Datta, M., Seano, G. et al. In vivo compression and imaging in mouse brain to measure the effects of solid stress. Nat Protoc 15, 2321–2340 (2020). https://doi.org/10.1038/s41596-020-0328-2

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