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High levels of TIMP1 are associated with increased extracellular matrix stiffness in isocitrate dehydrogenase 1-wild type gliomas

Abstract

Glioma progression is accompanied with increased tumor tissue stiffness, yet the underlying mechanisms are unclear. Herein, we employed atomic force microscopy analysis to show that tissue stiffness was higher in isocitrate dehydrogenase (IDH)-wild type gliomas than IDH-mutant gliomas. Bioinformatic analyses revealed that tissue inhibitor of metalloproteinase-1 (TIMP1) was one of the preferentially upregulated genes in IDH-wild type gliomas as compared to IDH-mutant gliomas, and its higher expression indicated worse prognosis of glioma patients. TIMP1 intensity determined by immunofluorescence staining on glioma tissues positively correlated with glioma tissue stiffness. Mechanistically, TIMP1 expression was positively correlated with the gene expression of two predominant extracellular matrix components, tenascin C and fibronectin, both of which were also highly expressed in IDH-wild type gliomas. By introducing IDH1-R132H-containing vectors into human IDH1-wild type glioma cells to obtain an IDH1-mutant cell line, we found that IDH1 mutation increased the TIMP1 promoter methylation through methylation-specific PCR. More importantly, IDH1-R132H mutation decreased both the expression of TIMP1, fibronectin, tenascin C, and the tumor tissue stiffness in IDH1-mutant glioma xenografts in contrast to IDH1-wild type counterparts. Moreover, TIMP1 knockdown in IDH-wild type glioma cells inhibited the expression of tenascin C and fibronectin, and decreased tissue stiffness in intracranial glioma xenografts. Conclusively, we revealed an IDH mutation status-mediated mechanism in regulating glioma tissue stiffness through modulating TIMP1 and downstream extracellular matrix components.

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Fig. 1: IDH-WT gliomas showed increased stiffness compared to IDH-Mut gliomas.
Fig. 2: TIMP1 was highly expressed in IDH-WT gliomas and predicted poor prognosis.
Fig. 3: TIMP1 is positively correlated with tissue stiffness and stiffness-related molecules.
Fig. 4: High expression of TIMP1 affects the tissue stiffness in glioma orthotopic xenografts.

Data availability

The authors confirm that the data supporting the findings of this study are available within the article or its supplementary materials.

References

  1. Zhang, L., He, L., Lugano, R., Roodakker, K., Bergqvist, M., Smits, A. et al. IDH mutation status is associated with distinct vascular gene expression signatures in lower-grade gliomas. Neuro Oncol 20, 1505–1516 (2018).

  2. Louis, D. N., Perry, A., Wesseling, P., Brat, D. J., Cree, I. A., Figarella-Branger, D. et al. The 2021 WHO Classification of Tumors of the Central Nervous System: A summary. Neuro Oncol 23, 1231–1251 (2021).

  3. Ng, H., Wong, Q. H., Liu, E. M., Li, K. K. The new WHO molecular criteria for adult glioblastoma – Are we a step too far? Glioma 4, 65–67 (2021).

  4. Yang, R., Zhao, Y., Gu, Y., Yang, Y., Gao, X., Yuan, Y. et al. Isocitrate dehydrogenase 1 mutation enhances 24(S)-hydroxycholesterol production and alters cholesterol homeostasis in glioma. Oncogene 39, 6340–6353 (2020).

  5. Waitkus, M. S., Diplas, B. H., Yan, H. Biological role and therapeutic potential of IDH mutations in cancer. Cancer Cell 34, 186–195 (2018).

  6. Pirozzi, C. J., Yan, H. The implications of IDH mutations for cancer development and therapy. Nat Rev Clin Oncol 18, 645–661 (2021).

  7. Levental, K. R., Yu, H., Kass, L., Lakins, J. N., Egeblad, M., Erler, J. T. et al. Matrix crosslinking forces tumor progression by enhancing integrin signaling. Cell 139, 891–906 (2009).

  8. Bonnans, C., Chou, J., Werb, Z. Remodelling the extracellular matrix in development and disease. Nat Rev Mol Cell Biol 15, 786–801 (2014).

  9. Miroshnikova, Y. A., Mouw, J. K., Barnes, J. M., Pickup, M. W., Lakins, J. N., Kim, Y. et al. Tissue mechanics promote IDH1-dependent HIF1alpha-tenascin C feedback to regulate glioblastoma aggression. Nat Cell Biol 18, 1336–1345 (2016).

  10. Cox, T. R. The matrix in cancer. Nat Rev Cancer 21, 217–238 (2021).

  11. Unnikandam, V. S., Hwang, D., Correia, J., Bartlett, M. D., Schneider, I. C. Cancer cell migration in collagen-hyaluronan composite extracellular matrices. Acta Biomater 130, 183–198 (2021).

  12. Dou, J., Mao, S., Li, H., Lin, J. M. Combination stiffness gradient with chemical stimulation directs glioma cell migration on a microfluidic chip. Anal Chem 92, 892–898 (2020).

  13. Grunwald, B., Schoeps, B., Kruger, A. Recognizing the molecular multifunctionality and interactome of TIMP-1. Trends Cell Biol 29, 6–19 (2019).

  14. Zhao, D., Li, Q., Liu, M., Ma, W., Zhou, T., Xue, C. et al. Substrate stiffness regulated migration and invasion ability of adenoid cystic carcinoma cells via RhoA/ROCK pathway. Cell Prolif 51, e12442 (2018).

  15. Guo, Y., Wang, X., Ning, W., Zhang, H., Yu, C. Identification of two core genes in glioblastomas with different isocitrate dehydrogenase mutation status. Mol Biol Rep 47, 7477–7488 (2020).

  16. Zhang, X. N., Yang, K. D., Chen, C., He, Z. C., Wang, Q. H., Feng, H. et al. Pericytes augment glioblastoma cell resistance to temozolomide through CCL5-CCR5 paracrine signaling. Cell Res 31, 1072–1087 (2021).

  17. Frantz, C., Stewart, K. M., Weaver, V. M. The extracellular matrix at a glance. J Cell Sci 123, 4195–4200 (2010).

  18. Dragos, A., Kovacs, A. T. The peculiar functions of the bacterial extracellular matrix. Trends Microbiol 25, 257–266 (2017).

  19. Bangasser, B. L., Shamsan, G. A., Chan, C. E., Opoku, K. N., Tuzel, E., Schlichtmann, B. W. et al. Shifting the optimal stiffness for cell migration. Nat Commun 8, 15313 (2017).

  20. Osmulski, P., Mahalingam, D., Gaczynska, M. E., Liu, J., Huang, S., Horning, A. M. et al. Nanomechanical biomarkers of single circulating tumor cells for detection of castration resistant prostate cancer. Prostate 74, 1297–1307 (2014).

  21. Bunse, L., Pusch, S., Bunse, T., Sahm, F., Sanghvi, K., Friedrich, M. et al. Suppression of antitumor T cell immunity by the oncometabolite (R)-2-hydroxyglutarate. Nat Med 24, 1192–1203 (2018).

  22. Chen, X., Wanggou, S., Bodalia, A., Zhu, M., Dong, W., Fan, J. J. et al. A feedforward mechanism mediated by mechanosensitive ion channel PIEZO1 and tissue mechanics promotes glioma aggression. Neuron 100, 799–815 (2018).

  23. Marhuenda, E., Fabre, C., Zhang, C., Martin-Fernandez, M., Iskratsch, T., Saleh, A. et al. Glioma stem cells invasive phenotype at optimal stiffness is driven by MGAT5 dependent mechanosensing. J Exp Clin Cancer Res 40, 139 (2021).

  24. Fan, Y., Sun, Q., Li, X., Feng, J., Ao, Z., Li, X. et al. Substrate stiffness modulates the growth, phenotype, and chemoresistance of ovarian cancer cells. Front Cell Dev Biol 9, 718834 (2021).

  25. Shen, Y., Wang, X., Lu, J., Salfenmoser, M., Wirsik, N. M., Schleussner, N. et al. Reduction of liver metastasis stiffness improves response to bevacizumab in metastatic colorectal cancer. Cancer Cell 37, 800–817 (2020).

  26. Medina, S. H., Bush, B., Cam, M., Sevcik, E., DelRio, F. W., Nandy, K. et al. Identification of a mechanogenetic link between substrate stiffness and chemotherapeutic response in breast cancer. Biomaterials 202, 1–11 (2019).

  27. Mohammadi, H. Sahai, E. Mechanisms and impact of altered tumour mechanics. Nat Cell Biol 20, 766–774 (2018).

  28. Dong, Y., Zheng, Q., Wang, Z., Lin, X., You, Y., Wu, S. et al. Higher matrix stiffness as an independent initiator triggers epithelial-mesenchymal transition and facilitates HCC metastasis. J Hematol Oncol 12, 112 (2019).

  29. Rouviere, O., Melodelima, C., Hoang, D. A., Bratan, F., Pagnoux, G., Sanzalone, T. et al. Stiffness of benign and malignant prostate tissue measured by shear-wave elastography: A preliminary study. Eur Radiol 27, 1858–1866 (2017).

  30. Amos, S. E., Choi, Y. S. The cancer microenvironment: Mechanical challenges of the metastatic cascade. Front Bioeng Biotechnol 9, 625859 (2021).

  31. Karamanos, N. K., Piperigkou, Z., Passi, A., Gotte, M., Rousselle, P., Vlodavsky, I. Extracellular matrix-based cancer targeting. Trends Mol Med 27, 1000–1013 (2021).

  32. Jackson, H. W., Defamie, V., Waterhouse, P., Khokha, R. TIMPs: Versatile extracellular regulators in cancer. Nat Rev Cancer 17, 38–53 (2017).

  33. Handsley, M. M., Edwards, D. R. Metalloproteinases and their inhibitors in tumor angiogenesis. Int J Cancer 115, 849–860 (2005).

  34. Fingleton, B. Matrix metalloproteinases: Roles in cancer and metastasis. Front Biosci 11, 479–491 (2006).

  35. Shi, F., Sottile, J. MT1-MMP regulates the turnover and endocytosis of extracellular matrix fibronectin. J Cell Sci 124, 4039–4050 (2011).

  36. Brosicke, N. & Faissner, A. Role of tenascins in the ECM of gliomas. Cell Adh Migr 9, 131–140 (2015).

  37. Saito, Y., Imazeki, H., Miura, S., Yoshimura, T., Okutsu, H., Harada, Y., et al. A peptide derived from tenascin-C induces beta1 integrin activation through syndecan-4. J Biol Chem 282, 34929–34937 (2007).

  38. Echtermeyer, F., Streit, M., Wilcox-Adelman, S., Saoncella, S., Denhez, F., Detmar, M. et al. Delayed wound repair and impaired angiogenesis in mice lacking syndecan-4. J Clin Invest 107, R9–R14 (2001).

  39. Dang, L., Su, S. M. Isocitrate dehydrogenase mutation and (R)-2-Hydroxyglutarate: From basic discovery to therapeutics development. Annu Rev Biochem 86, 305–331 (2017).

  40. Ricca, T. I., Liang, G., Suenaga, A. P., Han, S. W., Jones, P. A., Jasiulionis, M. G. Tissue inhibitor of metalloproteinase 1 expression associated with gene demethylation confers anoikis resistance in early phases of melanocyte malignant transformation. Transl Oncol 2, 329–340 (2009).

  41. Zhang, W., Sun, W., Qin, Y., Wu, C., He, L., Zhang, T. et al. Knockdown of KDM1A suppresses tumour migration and invasion by epigenetically regulating the TIMP1/MMP9 pathway in papillary thyroid cancer. J Cell Mol Med 23, 4933–4944 (2019).

  42. Wang, H., Liu, Z., Zhang, G. FBN1 promotes DLBCL cell migration by activating the Wnt/beta-catenin signaling pathway and regulating TIMP1. Am J Transl Res 12, 7340–7353 (2020).

  43. Huang, S. M., Wu, C. S., Chiu, M. H., Wu, C. H., Chang, Y. T., Chen, G. S. et al. High glucose environment induces M1 macrophage polarization that impairs keratinocyte migration via TNF-alpha: An important mechanism to delay the diabetic wound healing. J Dermatol Sci 96, 159–167 (2019).

  44. Hu, J., Shen, T., Xie, J., Wang, S., He, Y., Zhu, F. Curcumin modulates covalent histone modification and TIMP1 gene activation to protect against vascular injury in a hypertension rat model. Exp Ther Med 14, 5896–5902 (2017).

  45. Azevedo, M. J., Rabelo-Santos, S. H., Do, A. W. M., Zeferino, L. C. Tumoral and stromal expression of MMP-2, MMP-9, MMP-14, TIMP-1, TIMP-2, and VEGF-A in cervical cancer patient survival: A competing risk analysis. BMC Cancer 20, 660 (2020).

  46. Abreu, M., Cabezas-Sainz, P., Alonso-Alconada, L., Ferreiros, A., Mondelo-Macia, P., Lago-Leston, R. M. et al. Circulating tumor cells characterization revealed TIMP1 as a potential therapeutic target in ovarian cancer. Cells 9, 1218(2020).

  47. Barabas, L., Hritz, I., Istvan, G., Tulassay, Z., Herszenyi, L. The behavior of MMP-2, MMP-7, MMP-9, and their inhibitors TIMP-1 and TIMP-2 in Adenoma-Colorectal cancer sequence. Dig Dis 39, 217–224 (2021).

  48. Schoeps, B., Eckfeld, C., Prokopchuk, O., Bottcher, J., Haussler, D., Steiger, K. et al. TIMP1 triggers neutrophil extracellular trap formation in pancreatic cancer. Cancer Res 81, 3568–3579 (2021).

  49. Zhang, Z., Lv, J., Lei, X., Li, S., Zhang, Y., Meng, L. et al. Baicalein reduces the invasion of glioma cells via reducing the activity of p38 signaling pathway. PLoS One 9, e90318 (2014).

  50. Ando, T., Charindra, D., Shrestha, M., Umehara, H., Ogawa, I., Miyauchi, M. et al. Tissue inhibitor of metalloproteinase-1 promotes cell proliferation through YAP/TAZ activation in cancer. Oncogene 37, 263–270 (2018).

  51. Meng, Z., Qiu, Y., Lin, K. C., Kumar, A., Placone, J. K., Fang, C. et al. RAP2 mediates mechanoresponses of the Hippo pathway. Nature 560, 655–660 (2018).

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Funding

This research was supported by grants from the National Natural Science Foundation of China (81703012, 81902548), and Science and Technology Innovation Project of Chongqing Science and Technology Commission (cstc2017jcyj-yszxX0012).

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Conception and design: X.-W.B., Y.-F.P., and Y.S. Cell culture: C.-H.L. Staining: C.-H.L., Y.-Q.L., M.L., and J.-Y.M. Immune blotting: C.-H.L., W.-Y.W., H.Z., L.L. Animal experiments: C.-H.L. and M.M. MS-PCR: Q.L. Data analysis: C.C., Q.N., Z.-X.Y., K.-D.Y., and X.-N.Z. Writing of the manuscript: C.-H.L., Y.-F.P., and Y.S.

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Correspondence to Xiu-Wu Bian or Yi-Fang Ping.

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The stiffness of human glioma section measuring and human glioma section staining was approved by the Ethics Committees of Southwest Hospital (KY2021041). The animal use was approved by the Ethics Committees of Southwest Hospital (AMUWEC20210594).

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Luo, CH., Shi, Y., Liu, YQ. et al. High levels of TIMP1 are associated with increased extracellular matrix stiffness in isocitrate dehydrogenase 1-wild type gliomas. Lab Invest (2022). https://doi.org/10.1038/s41374-022-00825-4

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  • DOI: https://doi.org/10.1038/s41374-022-00825-4

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