Article | Published:

Molecular Diagnostics

Low tumour PPM1H indicates poor prognosis in colorectal cancer via activation of cancer-associated fibroblasts

Abstract

Background

Vimentin (VIM) is considered a prognostic marker in colorectal cancer (CRC). Our aim is to identify genes that fulfil a “X-low implies VIM-high” Boolean relationship and to evaluate their prognostic value and potential mechanism.

Methods

Potential biomarkers related to VIM expression were searched using a bioinformatics approach across gene-expression arrays. Based on subgroup analysis of 2 CRC cohorts, the selected gene was tested for its association with patient’s survival outcomes. The regulatory link between the selected gene and VIM was further examined with in vitro models.

Results

PPM1H was identified as the top candidate in our search. Patients with PPM1H-low tumours have a lower 5-year disease-free survival rate than patients with PPM1H-high tumours in 2 independent cohorts. In multivariate Cox analysis, patients with PPM1H-low tumours were independently associated with relapse in both the discovery cohort (hazard ratio [HR], 1.362; 95% confidence interval [CI], 1.015–1.826; P = 0.039) and the validation cohort (HR for DFS, 4.052; 95% CI, 2.634–6.234; P < 0.001). PPM1H knockdown in CRC cells and growth in the corresponding conditional medium increased VIM expression and colon fibroblast proliferation, indicating a transformation of cancer-association fibroblasts (CAFs). Conversely, educated CAFs also facilitated the growth of CRC cells with low PPM1H expression.

Conclusions

Lack of tumour PPM1H expression identifies a patient subgroup with a high relapse risk, and CRC cells with low expression of PPM1H activate CAFs and inversely get promoted by CAFs.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Bray F., Ferlay J., Soerjomataram I., Siegel R. L., Torre L. A., Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2018; e-pub ahead of print 12 Sep 2018; https://doi.org/10.3322/caac.21492.

  2. 2.

    Schmoll, H. J., V an Cutsem, E., Stein, A., Valentini, V., Glimelius, B. & Hausternams, K. et al. ESMO Consensus Guidelines for management of patients with colon and rectal cancer. a personalized approach to clinical decision making. Ann Oncol 23, 2479–2516 (2012).

  3. 3.

    Chang, W., Gao, X., Han, Y., Du, Y., Liu, Q. & Wang, L. et al. Gene expression profiling-derived immunohistochemistry signature with high prognostic value in colorectal carcinoma. Gut 63, 1457–1467 (2014).

  4. 4.

    Van Schaeybroeck, S., Allen, W. L., Turkington, R. C. & Johnston, P. G. Implementing prognostic and predictive biomarkers in CRC clinical trials. Nat Rev Clin Oncol 8, 222–232 (2011).

  5. 5.

    De Sousa, E., Melo, F., Wang, X., Jansen, M., Fessler, E., Trinh, A. & de Rooij, L. P. et al. Poor-prognosis colon cancer is defined by a molecularly distinct subtype and develops from serrated precursor lesions. Nat Med 19, 614–618 (2013).

  6. 6.

    Sadanandam, A., Lyssiotis, C. A., Homicsko, K., Collisson, E. A., Gibb, W. J. & Wullschleger, S. et al. A colorectal cancer classification system that associates cellular phenotype and responses to therapy. Nat Med 19, 619–625 (2013).

  7. 7.

    Calon, A., Lonardo, E., Berenguer-Llergo, A., Espinet, E., Hernando-Momblona, X. & Iglesias, M. et al. Stromal gene expression defines poor-prognosis subtypes in colorectal cancer. Nat Genet 47, 320–329 (2015).

  8. 8.

    Guinney, J., Dienstmann, R., Wang, X., de Reyniès, A., Schlicker, A. & Soneson, C. et al. The consensus molecular subtypes of colorectal cancer. Nat Med 21, 1350–1356 (2015).

  9. 9.

    Isella, C., Terrasi, A., Bellomo, S. E., Petti, C., Galatola, G. & Muratore, A. et al. Stromal contribution to the colorectal cancer transcriptome. Nat Genet 47, 312–319 (2015).

  10. 10.

    Li, H., Courtois, E. T., Sengupta, D., Tan, Y., Chen, K. H. & Goh, J. J. L. et al. Reference component analysis of single-cell transcriptomes elucidates cellular heterogeneity in human colorectal tumors. Nat Genet 49, 708–718 (2017).

  11. 11.

    Dienstmann, R., Vermeulen, L., Guinney, J., Kopetz, S., Tejpar, S. & Tabernero, J. Consensus molecular subtypes and the evolution of precision medicine in colorectal cancer. Nat Rev Cancer 17, 268 (2017).

  12. 12.

    Koliaraki, V., Pallangyo, C. K., Greten, F. R. & Kolias, G. Mesenchymal Cells in Colon Cancer. Gastroenterology 152, 964–979 (2017).

  13. 13.

    Kalluri, R. The biology and function of fibroblasts in cancer. Nat Rev Cancer 16, 582–598 (2016).

  14. 14.

    Fischer, K. R., Durrans, A., Lee, S., Sheng, J., Li, F. & Wong, S. T. et al. Epithelial-to-mesenchymal transition is not required for lung metastasis but contributes to chemoresistance. Nature 527, 472–476 (2015).

  15. 15.

    De Craene, B. & Berx, G. Regulatory networks defining EMT during cancer initiation and Progression. Nat Rev Cancer 13, 97–110 (2013).

  16. 16.

    Shi, Y., Du, L., Lin, L. & Wang, Y. Tumour-associated mesenchymal stem/stromal cells: emerging therapeutic targets. Nat Rev Drug 16, 35–52 (2017).

  17. 17.

    Toiyama, Y., Yasuda, H., Saigusa, S., Tanaka, K., Inoue, Y. & Goel, A. et al. Increased expression of Slug and Vimentin as novel predictive biomarkers for lymph node metastasis and poor prognosis in colorectal cancer. Carcinogenesis 34, 2548–2557 (2013).

  18. 18.

    Ngan, C. Y., Yamamoto, H., Seshimo, I., Tsujino, T., Man-i, M. & Ikeda, J. I. et al. Quantitative evaluation of vimentin expression in tumour stroma of colorectal cancer. Br J Cancer 96, 986–992 (2007).

  19. 19.

    Sahoo, D., Dill, D. L., Gentles, A. J., Tibshirani, R. & Plevritis, S. K. Boolean implication networks derived from large scale, whole genome microarray datasets. Genome Biol 9, R157 (2008).

  20. 20.

    Dalerba, P., Kalisky, T., Sahoo, D., Rajendran, P. S., Rothenberg, M. E. & Leyrat, A. A. et al. Single-cell dissection of transcriptional heterogeneity in human colon tumors. Nat Biotechnol 29, 1120–1127 (2011).

  21. 21.

    Dalerba, P., Sahoo, D., Paik, S., Guo, X., Yothers, G. & Song, N. et al. CDX2 as a Prognostic Biomarker in Stage II and Stage III Colon Cancer. N Engl J Med 374, 211–222 (2016).

  22. 22.

    Huo, Y., Su, T., Cai, Q. & Macara, I. G. An In Vivo Gain-of-Function Screen Identifies the Williams-Beuren Syndrome Gene GTF2IRD1 as a Mammary Tumor Promoter. Cell Rep 15, 2089–2096 (2016).

  23. 23.

    Chevillard, G., Paquet, M. & Blank, V. Nfe2l3 (Nrf3) deficiency predisposes mice to T-cell lymphoblastic lymphoma. Blood 117, 2005–2008 (2011).

  24. 24.

    Zhu, H., Qin, H., Li, D. M., Liu, J. & Zhao, Q. Effect of PPM1H on malignant phenotype of human pancreatic cancer cells. Oncol Rep 36, 2926–2934 (2016).

  25. 25.

    Shen, T., Sun, C., Zhang, Z., Xu, N., Duan, X. & Feng, X. H. et al. Specific control of BMP signaling and mesenchymal differentiation by cytoplasmic phosphatase PPM1H. Cell Res 24, 727–741 (2014).

  26. 26.

    Lee-Hoeflich, S. T., Pham, T. Q., Dowbenko, D., Munroe, X., Lee, J. & Li, L. et al. PPM1H is a p27 phosphatase implicated in trastuzumab resistance. Cancer Discov 1, 326–337 (2011).

  27. 27.

    Zhao, Y., Li, Q., Wu, X. & Chen, P. Upregulation of p27Kip1 by demethylation sensitizes cisplatin-resistant human ovarian cancer SKOV3 cells. Mol Med Rep 14, 1659–1666 (2016).

  28. 28.

    McCall, M. N., Bolstad, B. M. & Irizarry, R. A. Frozen robust multiarray analysis (fRMA). Biostatistics 11, 242–253 (2010).

Download references

Acknowledgements

We would like to acknowledge Association Dr Hanlin Tang, Department of Microbiology, Second Military Medical University, Shanghai for providing bioinformatics assistance. We thank Prof Dr Xiangyang Xue from the Department of Microbiology and Immunology, Wenzhou Medical University, Zhejiang for their technical support. This work was supported by grants from the National Natural Science Foundation of China (81572451 to WC; 81402005 to FC; 81502416 to YY).

Author contributions

X.X, L.Z., and Y.Y. analysed the whole data independently and presented the same results. F.C. and J.S. were responsible for the follow-up of CRC patients; W.C. and Y.P. were responsible for pathological analysis. Z.F., Y.L., and J.S. involved in the pathological diagnosis and recruitment of the patients in the hospital. X.X., L.Z. and W.C. were responsible for statistical analysis. X.X., Y.Y. and F.C. designed and organised the study and wrote the manuscript.

Author information

Correspondence to Wenjun Chang or Jinke Sui or Fuao Cao.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent

The ethics committee of Changhai Hospital ruled that no formal ethics approval was required in this particular case. The study was performed in accordance with the Declaration of Helsinki.

Consent for publication

All authors agree to publish the paper in its present form.

Data availability

All the data analysed or generated in this study are included in this article and its supplementary information file.

Note

This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Data Supplement

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark
Fig. 1
Fig. 2
Fig. 3
Fig. 4