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Quantifying solid stress and elastic energy from excised or in situ tumors

Abstract

Solid stress, distinct from both tissue stiffness and fluid pressure, is a mechanical stress that is often elevated in both murine and human tumors. The importance of solid stress in tumor biology has been recognized in initial studies: solid stress promotes tumor progression and lowers the efficacy of anticancer therapies by compressing blood vessels and contributing to hypoxia. However, robust, reproducible, and objective methods that go beyond demonstration and bulk measurements have not yet been established. We have developed three new techniques to rigorously measure and map solid stress in both human and murine tumors that are able to account for heterogeneity in the tumor microenvironment. We describe here these methods and their independent advantages: 2D spatial mapping of solid stress (planar-cut method), sensitive estimation of solid stress in small tumors (slicing method), and in situ solid-stress quantification (needle-biopsy method). Furthermore, the preservation of tissue morphology and structure allows for subsequent histological analyses in matched tumor sections, facilitating quantitative correlations between solid stress and markers of interest. The three procedures each require 2 h of experimental time per tumor. The required skill sets include basic experience in tumor resection and/or biopsy (in mice or humans), as well as in intravital imaging (e.g., ultrasonography).

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Figure 1: Overview of the procedure for estimation of solid stress in tumors.
Figure 2: Planar-cut method.
Figure 3: Slicing method.
Figure 4: Needle-biopsy method.
Figure 5: Indentation modulus at the tumor periphery versus the core.
Figure 6: Comparison between planar-cut and needle-biopsy methods.

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Acknowledgements

We thank S. Roberge for technical assistance. We thank N. Bardeesy and T. Irimura for providing AK4.4 and SL4 cells, respectively. This work was supported in part by the National Cancer Institute (P01-CA080124, R35-CA197743, and R01-CA208205 to R.K.J.); a fellowship from the National Cancer Institute (F32-CA216944-01 to H.T.N.); a fellowship from the Susan G. Komen Foundation (PDF14201739 to G.S.); and a fellowship from the National Heart, Lung, and Blood Institute (F31HL126449 to M.D.).

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Contributions

H.T.N., M.D., and R.K.J. designed the study; H.T.N. and M.D. were responsible for acquisition of the data. H.T.N., M.D., G.S., P.H., L.L.M., and R.K.J. contributed to analysis and interpretation of the data. H.T.N., M.D., G.S., P.H., L.L.M., and R.K.J. were involved in drafting of 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. has received consultant fees from Enlight, Merck, Ophthotech, Pfizer, SPARC, and SynDevRx; owns equity in Enlight, Ophthotech, SynDevRx, and XTuit; and serves on the Board of Directors of XTuit and 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. The other authors declare no competing interests.

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Nia, H., Datta, M., Seano, G. et al. Quantifying solid stress and elastic energy from excised or in situ tumors. Nat Protoc 13, 1091–1105 (2018). https://doi.org/10.1038/nprot.2018.020

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