Abstract
Hyaluronan is a major component of the extracellular matrix in both normal and tumor tissue. Many solid cancers, including bladder cancer, are characterized by deregulated hyaluronan metabolism. It is postulated that the deregulated metabolism in cancer tissue is characterized by elevated hyaluronan synthesis and degradation. This results in the accumulation of small hyaluronan fragments in the tumor microenvironment which promotes cancer-related inflammation, stimulates tumor cell proliferation and angiogenesis, and contributes to immune-associated immune suppression. For a better understanding of the complex mechanisms of hyaluronan metabolism in cancer, it has been proposed to use precision-cut tissue slice cultures prepared using freshly excised cancer tissue. Here we describe the protocol for establishing tissue slice cultures and analysis of tumor-associated hyaluronan in human urothelial carcinoma.
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References
Girish KS, Kemparaju K (2007) The magic glue hyaluronan and its eraser hyaluronidase: a biological overview. Life Sci 80:1921–1943. https://doi.org/10.1016/j.lfs.2007.02.037
Toole BP (2004) Hyaluronan: from extracellular glue to pericellular cue. Nat Rev Cancer 4:528–539. https://doi.org/10.1038/nrc1391
Sironen RK, Tammi M, Tammi R et al (2011) Hyaluronan in human malignancies. Exp Cell Res 317:383–391. https://doi.org/10.1016/j.yexcr.2010.11.017
Itano N, Atsumi F, Sawai T et al (2002) Abnormal accumulation of hyaluronan matrix diminishes contact inhibition of cell growth and promotes cell migration. Proc Natl Acad Sci U S A 99:3609–3614. https://doi.org/10.1073/pnas.052026799
Voelcker V, Gebhardt C, Averbeck M et al (2008) Hyaluronan fragments induce cytokine and metalloprotease upregulation in human melanoma cells in part by signalling via TLR4. Exp Dermatol 17:100–107. https://doi.org/10.1111/j.1600-0625.2007.00638.x
Sokolowska M, Chen L-Y, Eberlein M et al (2014) Low molecular weight hyaluronan activates cytosolic phospholipase A2α and eicosanoid production in monocytes and macrophages. J Biol Chem 289:4470–4488. https://doi.org/10.1074/jbc.M113.515106
Donelan W, Dominguez-Gutierrez PR, Kusmartsev S (2022) Deregulated hyaluronan metabolism in the tumor microenvironment drives cancer inflammation and tumor-associated immune suppression. Front Immunol 13:971278. https://doi.org/10.3389/fimmu.2022.971278
Liu N, Gao F, Han Z et al (2001) Hyaluronan synthase 3 overexpression promotes the growth of TSU prostate cancer cells. Cancer Res 61:5207–5214
Schmaus A, Bauer J, Sleeman JP (2014) Sugars in the microenvironment: the sticky problem of HA turnover in tumors. Cancer Metastasis Rev 33:1059–1079. https://doi.org/10.1007/s10555-014-9532-2
Turley EA, Wood DK, McCarthy JB (2016) Carcinoma cell hyaluronan as a “portable” cancerized prometastatic microenvironment. Cancer Res 76:2507–2512. https://doi.org/10.1158/0008-5472.CAN-15-3114
Dominguez-Gutierrez PR, Kwenda EP, Donelan W et al (2022) Detection of PD-L1-expressing myeloid cell clusters in the hyaluronan-enriched stroma in tumor tissue and tumor-draining lymph nodes. J Immunol 208:2829–2836. https://doi.org/10.4049/jimmunol.2100026
Stern R (2008) Hyaluronidases in cancer biology. Semin Cancer Biol 18:275–280. https://doi.org/10.1016/j.semcancer.2008.03.017
Kramer MW, Golshani R, Merseburger AS et al (2010) HYAL-1 hyaluronidase: a potential prognostic indicator for progression to muscle invasion and recurrence in bladder cancer. Eur Urol 57:86–93. https://doi.org/10.1016/j.eururo.2009.03.057
Harada H, Takahashi M (2007) CD44-dependent intracellular and extracellular catabolism of hyaluronic acid by hyaluronidase-1 and -2. J Biol Chem 282:5597–5607. https://doi.org/10.1074/jbc.M608358200
Dominguez-Gutierrez PR, Kwenda EP, Donelan W et al (2021) Hyal2 expression in tumor-associated myeloid cells mediates cancer-related inflammation in bladder cancer. Cancer Res 81:648–657. https://doi.org/10.1158/0008-5472.CAN-20-1144
Acknowledgments
We would like to thank Paul R. Dominguez-Gutierrez (currently at Resilience, Inc.) and Elizabeth Kwenda (University of Florida) for assisting to establish the tissue slice protocol in the lab. We also would like to acknowledge the role of Padraic O’Malley (University of Florida) in providing tissue samples from cancer patients.
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Donelan, W., Crispen, P.L., Kusmartsev, S. (2023). Tissue Slice Culture and Analysis of Tumor-Associated Hyaluronan in Urothelial Carcinoma. In: Hoffmann, M.J., Gaisa, N.T., Nawroth, R., Ecke, T.H. (eds) Urothelial Carcinoma. Methods in Molecular Biology, vol 2684. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3291-8_10
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DOI: https://doi.org/10.1007/978-1-0716-3291-8_10
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