A stromal cell population in the large intestine identified by tissue factor expression that is lost during colorectal cancer progression

Summary

Colorectal cancer (CRC) is a major cause of morbidity and mortality, and the composition of the tumour stroma is a strong predictor of survival in this cancer type. Tissue factor (TF) functions as the trigger of haemostasis together with its ligand coagulation factor VII/VIIa, and TF expression has been found in tumour cells of colorectal tumours. However, TF expression in the CRC tumour stroma or its relationship to patient outcome has not yet been studied. To address this question we developed and validated a specific anti-TF antibody using standardised methods within the Human Protein Atlas project. We used this antibody to investigate TF expression in normal colorectal tissue and CRC using immunofluorescence and immunohistochemistry in two patient cohorts. TF was strongly expressed in a cell population immediately adjacent to the colorectal epithelium. These TF-positive cells were ACTA2-negative but weakly vimentin-positive, defining a specific population of pericryptal sheath cells. In colorectal tumours, TF-positive sheath cells were progressively lost after the adenoma-to-carcinoma transition, demonstrating downregulation of this source of TF in CRC. Furthermore, loss of sheath cell TF was significantly associated with poor overall and disease-specific survival in rectal but not colon cancers. In conclusion, we demonstrate that TF is a marker of a specific cell population in the large intestine, which is lost during CRC progression. Our results highlight the role of the tumour stroma in this cancer type and suggest TF to be a potential prognostic biomarker in rectal cancers through the identification of pericryptal sheath cells.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Tissue microarray - Tissue Factor/factor VII - immunohistochemistry - biomarker - colorectal cancer

• References

• 1 Ferlay J, Soerjomataram I, Dikshit R. et al. Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 2015; 136: E359-386.
• 2 Roth AD, Delorenzi M, Tejpar S. et al. Integrated Analysis of Molecular and Clinical Prognostic Factors in Stage II/III Colon Cancer. J Natl Cancer Inst 2012; 104: 1635-1646.
• 3 Sorbye H, Dragomir A, Sundström M. et al. High BRAF Mutation Frequency and Marked Survival Differences in Subgroups According to KRAS/BRAF Mutation Status and Tumour Tissue Availability in a Prospective Population-Based Metastatic Colorectal Cancer Cohort. PLoS ONE 2015; 10: e0131046.
• 4 Åberg M, Eriksson O, Siegbahn A. Tissue Factor Noncoagulant Signalling: Mechanisms and Implications for Cell Migration and Apoptosis. Semin Thromb Haemost 2015; 41: 691-699.
• 5 Eriksson O, Ramström M, Hörnaeus K. et al. The Eph Tyrosine Kinase Receptors EphB2 and EphA2 Are Novel Proteolytic Substrates of Tissue Factor/Coagulation Factor VIIa. J Biol Chem 2014; 289: 32379-32391.
• 6 Drake TA, Morrissey JH, Edgington TS. Selective cellular expression of tissue factor in human tissues. Implications for disorders of haemostasis and thrombosis. Am J Pathol 1989; 134: 1087-1097.
• 7 Fleck RA, Rao LVM, Rapaport SI. et al. Localisation of human tissue factor antigen by immunostaining with monospecific, polyclonal anti-human tissue factor antibody. Thromb Res 1990; 59: 421-437.
• 8 Rao LVM, Kothari H, Pendurthi UR. Tissue factor: mechanisms of decryption. Front Biosci Elite Ed 2012; 04: 1513-1527.
• 9 Calon A, Lonardo E, Berenguer-Llergo A. et al. Stromal gene expression defines poor-prognosis subtypes in colorectal cancer. Nat Genet 2015; 47: 320-329.
• 10 Kocatürk B, Berg den YWV, Tieken C. et al. Alternatively spliced tissue factor promotes breast cancer growth in a β1 integrin-dependent manner. Proc Natl Acad Sci USA 2013; 110: 11517-11522.
• 11 Hamada K, Kuratsu J, Saitoh Y. et al. Expression of tissue factor correlates with grade of malignancy in human glioma. Cancer 1996; 77: 1877-1883.
• 12 Rao B, Gao Y, Huang J. et al. Mutations of p53 and K-ras correlate TF expression in human colorectal carcinomas: TF downregulation as a marker of poor prognosis. Int J Colorectal Dis 2011; 26: 593-601.
• 13 Uhlén M, Fagerberg L, Hallström BM. et al. Tissue-based map of the human proteome. Science 2015; 347: 1260419.
• 14 Nilsson P, Paavilainen L, Larsson K. et al. Towards a human proteome atlas: High-throughput generation of mono-specific antibodies for tissue profiling. Proteomics 2005; 05: 4327-4337.
• 15 O’Hurley G, Sjöstedt E, Rahman A. et al. Garbage in, garbage out: A critical evaluation of strategies used for validation of immunohistochemical biomarkers. Mol Oncol 2014; 08: 783-798.
• 16 Kampf C, Olsson I, Ryberg U. et al. Production of Tissue Microarrays, Immu-nohistochemistry Staining and Digitalisation Within the Human Protein Atlas. J Vis Exp [Internet] 2012 [cited 2016 Jan 25]. Available from http://www.jove.com/video/3620/
• 17 Hansen S, Sørensen FB, Vach W. et al. Microvessel density compared with the Chalkley count in a prognostic study of angiogenesis in breast cancer patients. Histopathology 2004; 44: 428-436.
• 18 Fagerberg L, Strömberg S, El-Obeid A. et al. Large-Scale Protein Profiling in Human Cell Lines Using Antibody-Based Proteomics. J Proteome Res 2011; 10: 4066-4075.
• 19 Mifflin RC, Pinchuk IV, Saada JI. et al. Intestinal myofibroblasts: targets for stem cell therapy. Am J Physiol - Gastrointest Liver Physiol 2011; 300: G684-696.
• 20 Eriksson O, Thulin Å, Asplund A. et al. Cross-talk between the Tissue Factor/ coagulation factor VIIa complex and the tyrosine kinase receptor EphA2 in cancer. BMC Cancer 2016; 16: 341.
• 21 Nakasaki T, Wada H, Shigemori C. et al. Expression of tissue factor and vascular endothelial growth factor is associated with angiogenesis in colorectal cancer. Am J Hematol 2002; 69: 247-254.
• 22 Kurahashi M, Nakano Y, Peri LE. et al. A novel population of subepithelial platelet-derived growth factor receptor α-positive cells in the mouse and human colon. Am J Physiol Gastrointest Liver Physiol 2013; 304: G823-834.
• 23 Powell DW, Adegboyega PA, Di Mari JF. et al. Epithelial Cells and Their Neighbors I. Role of intestinal myofibroblasts in development, repair, and cancer. Am J Physiol - Gastrointest Liver Physiol 2005; 289: G2-7.
• 24 Tsujino T, Seshimo I, Yamamoto H. et al. Stromal Myofibroblasts Predict Disease Recurrence for Colorectal Cancer. Clin Cancer Res 2007; 13: 2082-2090.
• 25 Berdiel-Acer M, Sanz-Pamplona R, Calon A. et al. Differences between CAFs and their paired NCF from adjacent colonic mucosa reveal functional heterogeneity of CAFs, providing prognostic information. Mol Oncol 2014; 08: 1290-1305.
• 26 Sidney LE, Branch MJ, Dunphy SE. et al. Concise Review: Evidence for CD34 as a Common Marker for Diverse Progenitors. Stem Cells Dayt Ohio 2014; 32: 1380-1389.
• 27 Carmeliet P, Mackman N, Moons L. et al. Role of tissue factor in embryonic blood vessel development. Nature 1996; 383: 73-75.
• 28 Arderiu G, Peña E, Aledo R. et al. Tissue Factor Regulates Microvessel Formation and Stabilisation by Induction of Chemokine (C-C Motif) Ligand 2 Expression. Arterioscler Thromb Vasc Biol 2011; 31: 2607-2615.
• 29 Yu J, May L, Milsom C. et al. Contribution of Host-Derived Tissue Factor to Tumour Neovascularisation. Arterioscler Thromb Vasc Biol 2008; 28: 1975-1981.
• 30 Ruf W, Disse J, Carneiro-Lobo TC. et al. Tissue factor and cell signalling in cancer progression and thrombosis. J Thromb Haemost JTH 2011; 09 (Suppl. 01) 306-315.
• 31 Tan Jr KK, de GLL, Sim R. How Uncommon are Isolated Lung Metastases in Colorectal Cancer? A Review from Database of 754 Patients Over 4 Years. J Gastrointest Surg 2009; 13: 642-648.
• 32 Kalady MF, Sanchez JA, Manilich E. et al. Divergent Oncogenic Changes Influence Survival Differences between Colon and Rectal Adenocarcinomas: Dis Colon Rectum. 2009; 52: 1039-1045.
• 33 Kapiteijn E, Liefers GJ, Los LC. et al. Mechanisms of oncogenesis in colon versus rectal cancer. J Pathol 2001; 195: 171-178.
• 34 Sanz-Pamplona R, Cordero D, Berenguer A. et al. Gene Expression Differences between Colon and Rectum Tumours. Clin Cancer Res 2011; 17: 7303-7312.
• 35 Berg VDWY, Osanto S. et al. The Relationship Between Tissue Factor and Cancer Progression: Insights from Bench and Bedside. Blood 2012; 119: 924-932.
• 36 Yu JL, May L, Lhotak V. et al. Oncogenic Events Regulate Tissue Factor Expression in Colorectal Cancer Cells: Implications for Tumour Progression and Angiogenesis. Blood 2005; 105: 1734-1741.
• 37 Prandoni P, Falanga A, Piccioli A. Cancer and venous thromboembolism. Lancet Oncol 2005; 06: 401-410.
• 38 Furie B, Furie BC. Cancer-associated thrombosis. Blood Cells Mol Dis 2006; 36: 177-181.
• 39 Cunningham MS, Preston RJS, O’Donnell JS. Does antithrombotic therapy improve survival in cancer patients?. Blood Rev 2009; 23: 129-135.
• 40 Tagalakis V, Blostein M, Robinson-Cohen C. et al. The effect of anticoagulants on cancer risk and survival: Systematic review. Cancer Treat Rev 2007; 33: 358-368.
• 41 Pengo V, Noventa F, Denas G. et al. Long-term use of vitamin K antagonists and incidence of cancer: a population-based study. Blood 2011; 117: 1707-1709.
• 42 Daly L. The first international urokinase/warfarin trial in colorectal cancer. Clin Exp Metastasis 1991; 09: 03-11.
• 43 Zacharski LR, Henderson WG, Rickles FR. et al. Effect of warfarin anticoagulation on survival in carcinoma of the lung, colon, head and neck, and prostate: Final Report of VA cooperative study # 75. Cancer 1984; 53: 2046-2052.