Quantification of tumour budding, lymphatic vessel density and invasion through image analysis in colorectal cancer

Journal of Translational Medicine

David J Harrison¹  Anca Oniscu¹  Susan M Farrington²  Arran K Turnbull²  Peter D Caie² 

[1] Systems Pathology, School of Medicine, University of St Andrews, St Andrews KY16 9TF, UK;Institute of Genetics and Molecular Medicine, University of Edinburgh, Western General Hospital, Edinburgh EH4 2XU, UK

关键词: Digital pathology;    Prognosis;    Colorectal;    Tumour budding;    Lymphatic vessel invasion;    Lymphatic vessel density;    Image analysis;   

Others  :  1148917
DOI  :  10.1186/1479-5876-12-156

 received in 2014-02-13, accepted in 2014-05-26,  发布年份 2014

【 摘 要 】

Background

Tumour budding (TB), lymphatic vessel density (LVD) and lymphatic vessel invasion (LVI) have shown promise as prognostic factors in colorectal cancer (CRC) but reproducibility using conventional histopathology is challenging. We demonstrate image analysis methodology to quantify the histopathological features which could permit standardisation across institutes and aid risk stratification of Dukes B patients.

Methods

Multiplexed immunofluorescence of pan-cytokeratin, D2-40 and DAPI identified epithelium, lymphatic vessels and all nuclei respectively in tissue sections from 50 patients diagnosed with Dukes A (n = 13), Dukes B (n = 29) and Dukes C (n = 8) CRC. An image analysis algorithm was developed and performed, on digitised images of the CRC tissue sections, to quantify TB, LVD, and LVI at the invasive front.

Results

TB (HR =5.7; 95% CI, 2.38-13.8), LVD (HR =5.1; 95% CI, 2.04-12.99) and LVI (HR =9.9; 95% CI, 3.57-27.98) were successfully quantified through image analysis and all were shown to be significantly associated with poor survival, in univariate analyses. LVI (HR =6.08; 95% CI, 1.17-31.41) is an independent prognostic factor within the study and was correlated to both TB (Pearson r =0.71, p <0.0003) and LVD (Pearson r =0.69, p <0.0003).

Conclusion

We demonstrate methodology through image analysis which can standardise the quantification of TB, LVD and LVI from a single tissue section while decreasing observer variability. We suggest this technology is capable of stratifying a high risk Dukes B CRC subpopulation and we show the three histopathological features to be of prognostic significance.

【 授权许可】

   
2014 Caie et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Standards and Datasets for Reporting Cancers Dataset for colorectal cancer. 2nd edition. 2007. [http://www.rcpath.org/Resources/RCPath/Migrated%20Resources/Documents/G/G049-ColorectalDataset-Sep07.pdf webcite]
• [2]Lan YT, Yang SH, Chang SC, Liang WY, Li AF, Wang HS, Jiang JK, Chen WS, Lin TC, Lin JK: Analysis of the seventh edition of American Joint Committee on colon cancer staging. Int J Colorectal Dis 2012, 27:657-663.
• [3]Poston GJ, Tait D, O'Connell S, Bennett A, Berendse S: Diagnosis and management of colorectal cancer: summary of NICE guidance. BMJ 2011, 343:d6751.
• [4]O'Connell JB, Maggard MA, Ko CY: Colon cancer survival rates with the new American Joint Committee on Cancer sixth edition staging. J Natl Cancer Inst 2004, 96:1420-1425.
• [5]Huh JW, Lee JH, Kim HR, Kim YJ: Prognostic significance of lymphovascular or perineural invasion in patients with locally advanced colorectal cancer. Am J Surg 2013, 206:758-763.
• [6]Chang S-C, Lin C-C, Wang H-S, Yang S-H, Jiang J-K, Lan Y-T, Lin T-C, Li AF-Y, Chen W-S, Lin J-K: Lymphovascular invasion determines the outcome of stage I colorectal cancer patients. Formosan J Surgery 2012, 45:141-145.
• [7]Betge J, Pollheimer MJ, Lindtner RA, Kornprat P, Schlemmer A, Rehak P, Vieth M, Hoefler G, Langner C: Intramural and extramural vascular invasion in colorectal cancer: prognostic significance and quality of pathology reporting. Cancer 2012, 118:628-638.
• [8]Barresi V, Reggiani-Bonetti L, Di Gregorio C, De Leon MP, Barresi G: Lymphatic vessel density and its prognostic value in stage I colorectal carcinoma. J Clin Pathol 2011, 64:6-12.
• [9]Cacchi C, Arnholdt HM, Jahnig H, Anthuber M, Probst A, Oruzio DV, Markl B: Clinical significance of lymph vessel density in T3 colorectal carcinoma. Int J Colorectal Dis 2012, 27:721-726.
• [10]Royston D, Jackson DG: Mechanisms of lymphatic metastasis in human colorectal adenocarcinoma. J Pathol 2009, 217:608-619.
• [11]Betge J, Kornprat P, Pollheimer MJ, Lindtner RA, Schlemmer A, Rehak P, Vieth M, Langner C: Tumour budding is an independent predictor of outcome in AJCC/UICC stage II colorectal cancer. Ann Surg Oncol 2012, 19:3706-3712.
• [12]Karamitopoulou E, Zlobec I, Kolzer V, Kondi-Pafiti A, Patsouris ES, Gennatas K, Lugli A: Proposal for a 10-high-power-fields scoring method for the assessment of tumour budding in colorectal cancer. Mod Pathol 2013, 26:295-301.
• [13]Ueno H, Murphy J, Jass JR, Mochizuki H, Talbot IC: Tumour 'budding' as an index to estimate the potential of aggressiveness in rectal cancer. Histopathology 2002, 40:127-132.
• [14]Mitrovic B, Schaeffer DF, Riddell RH, Kirsch R: Tumour budding in colorectal carcinoma: time to take notice. Mod Pathol 2012, 25:1315-1325.
• [15]Lugli A, Karamitopoulou E, Zlobec I: Tumour budding: a promising parameter in colorectal cancer. Br J Cancer 2012, 106:1713-1717.
• [16]Mou S, Soetikno R, Shimoda T, Rouse R, Kaltenbach T: Pathologic predictive factors for lymph node metastasis in submucosal invasive (T1) colorectal cancer: a systematic review and meta-analysis. Surg Endosc 2013, 27:2692-2703.
• [17]Glasgow SC, Bleier JI, Burgart LJ, Finne CO, Lowry AC: Meta-analysis of histopathological features of primary colorectal cancers that predict lymph node metastases. J Gastrointest Surg 2012, 16:1019-1028.
• [18]Uribarrena AR, Ortego J, Fuentes J, Raventos N, Parra P, Uribarrena ER: Prognostic value of microvascular density in dukes a and B (t1-t4, n0, m0) colorectal carcinomas. Gastroenterol Res Pract 2009, 2009:679830.
• [19]Suzuki A, Togashi K, Nokubi M, Koinuma K, Miyakura Y, Horie H, Lefor AT, Yasuda Y: Evaluation of venous invasion by Elastica van Gieson stain and tumour budding predicts local and distant metastases in patients with T1 stage colorectal cancer. Am J Surg Pathol 2009, 33:1601-1607.
• [20]Doyle B, Hagan S, Al-Mulla F, Scott L, Harden S, Paul J, Mulcahy H, Murray GI, Sheahan K, O'Sullivan J, Kolch W: Raf kinase inhibitor protein expression combined with peritoneal involvement and lymphovascular invasion predicts prognosis in Dukes' B colorectal cancer patients. Histopathology 2013, 62:505-510.
• [21]Betge J, Kornprat P, Pollheimer MJ, Lindtner RA, Schlemmer A, Rehak P, Vieth M, Langner C: Tumour Budding is an Independent Predictor of Outcome in AJCC/UICC Stage II Colorectal Cancer. Ann Surg Oncol 2012, 19:3706-3712.
• [22]Barresi V, Reggiani Bonetti L, Vitarelli E, Di Gregorio C, Ponz De Leon M, Barresi G: Immunohistochemical assessment of lymphovascular invasion in stage I colorectal carcinoma: prognostic relevance and correlation with nodal micrometastases. Am J Surg Pathol 2012, 36:66-72.
• [23]Puppa G, Senore C, Sheahan K, Vieth M, Lugli A, Zlobec I, Pecori S, Wang LM, Langner C, Mitomi H, Nakamura T, Watanabe M, Ueno H, Chasle J, Conley SA, Herlin P, Lauwers GY, Risio M: Diagnostic reproducibility of tumour budding in colorectal cancer: a multicentre, multinational study using virtual microscopy. Histopathology 2012, 61:562-575.
• [24]Horcic M, Koelzer VH, Karamitopoulou E, Terracciano L, Puppa G, Zlobec I, Lugli A: Tumour budding score based on 10 high-power fields is a promising basis for a standardised prognostic scoring system in stage II colorectal cancer. Hum Pathol 2013, 44:697-705.
• [25]Hasan J, Byers R, Jayson GC: Intra-tumoural microvessel density in human solid tumours. Br J Cancer 2002, 86:1566-1577.
• [26]Roxburgh CS, McMillan DC, Anderson JH, McKee RF, Horgan PG, Foulis AK: Elastica staining for venous invasion results in superior prediction of cancer-specific survival in colorectal cancer. Ann Surg 2010, 252:989-997.
• [27]Kingston EF, Goulding H, Bateman AC: Vascular invasion is underrecognized in colorectal cancer using conventional hematoxylin and eosin staining. Dis Colon Rectum 2007, 50:1867-1872.
• [28]Ohtsuki K, Koyama F, Tamura T, Enomoto Y, Fujii H, Mukogawa T, Nakagawa T, Uchimoto K, Nakamura S, Nonomura A, Nakajima Y: Prognostic value of immunohistochemical analysis of tumour budding in colorectal carcinoma. Anticancer Res 2008, 28:1831-1836.
• [29]Lin M, Ma SP, Lin HZ, Ji P, Xie D, Yu JX: Intratumoural as well as peritumoural lymphatic vessel invasion correlates with lymph node metastasis and unfavourable outcome in colorectal cancer. Clin Exp Metastasis 2010, 27:123-132.
• [30]Kazama S, Watanabe T, Ajioka Y, Kanazawa T, Nagawa H: Tumour budding at the deepest invasive margin correlates with lymph node metastasis in submucosal colorectal cancer detected by anticytokeratin antibody CAM5.2. Br J Cancer 2006, 94:293-298.
• [31]Tetu B, Evans A: Canadian Licensure for the Use of Digital Pathology for Routine Diagnoses. Arch Pathol Lab Med 2013, 138:302-304.
• [32]Pages F, Berger A, Camus M, Sanchez-Cabo F, Costes A, Molidor R, Mlecnik B, Kirilovsky A, Nilsson M, Damotte D, Meatchi T, Bruneval P, Cugnenc PH, Trajanoski Z, Fridman WH, Galon J: Effector memory T cells, early metastasis, and survival in colorectal cancer. N Engl J Med 2005, 353:2654-2666.
• [33]Angell HK, Gray N, Womack C, Pritchard DI, Wilkinson RW, Cumberbatch M: Digital pattern recognition-based image analysis quantifies immune infiltrates in distinct tissue regions of colorectal cancer and identifies a metastatic phenotype. Br J Cancer 2013, 109:1618-1624.
• [34]Hase K, Shatney C, Johnson D, Trollope M, Vierra M: Prognostic value of tumour "budding" in patients with colorectal cancer. Dis Colon Rectum 1993, 36:627-635.
• [35]Camp RL, Dolled-Filhart M, Rimm DL: X-tile: a new bio-informatics tool for biomarker assessment and outcome-based cut-point optimization. Clin Cancer Res 2004, 10:7252-7259.
• [36]Lubbock AL, Katz E, Harrison DJ, Overton IM: TMA Navigator: Network inference, patient stratification and survival analysis with tissue microarray data. Nucleic Acids Res 2013, 41:W562-W568.
• [37]Midgley R, Kerr DJ: Adjuvant chemotherapy for stage II colorectal cancer: the time is right! Nat Clin Pract Oncol 2005, 2:364-369.
• [38]Wang TB, Chen ZG, Wei XQ, Wei B, Dong WG: Serum vascular endothelial growth factor-C and lymphoangiogenesis are associated with the lymph node metastasis and prognosis of patients with colorectal cancer. ANZ J Surg 2011, 81:694-699.
• [39]Gao J, Knutsen A, Arbman G, Carstensen J, Franlund B, Sun XF: Clinical and biological significance of angiogenesis and lymphangiogenesis in colorectal cancer. Dig Liver Dis 2009, 41:116-122.
• [40]Longatto-Filho A, Pinheiro C, Ferreira L, Scapulatempo C, Alves VA, Baltazar F, Schmitt F: Peritumoural, but not intratumoural, lymphatic vessel density and invasion correlate with colorectal carcinoma poor-outcome markers. Virchows Arch 2008, 452:133-138.
• [41]Matsumoto K, Nakayama Y, Inoue Y, Minagawa N, Katsuki T, Shibao K, Tsurudome Y, Hirata K, Nagata N, Itoh H: Lymphatic microvessel density is an independent prognostic factor in colorectal cancer. Dis Colon Rectum 2007, 50:308-314.