Top

Journal of Experimental & Clinical Cancer Research

Published in:

Open Access 01-12-2014 | Research

In silico and Ex vivo approaches identify a role for toll-like receptor 4 in colorectal cancer

Authors: Daniel A Sussman, Rebeca Santaolalla, Pablo A Bejarano, Monica T Garcia-Buitrago, Maria T Perez, Maria T Abreu, Jennifer Clarke

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2014

Abstract

Background

Inflammation increases the risk of colorectal cancer (CRC). We and others have described a role for TLR4, the receptor for LPS, in colon cancer. To explore the relationships between TLR4 expression and CRC, we combined the strength of transcriptome array data and immunohistochemical (IHC) staining.

Methods

TLR4 signal intensity was scored in the stromal and epithelial compartments. Detection of differential expression between conditions of interest was performed using linear models, Cox proportional hazards models, and empirical Bayes methods.

Results

A strong association between TLR4 expression and survival was noted, though a dichotomous relationship between survival and specific TLR4 transcripts was observed. Increasing TLR4 expression was seen with advancing tumor stage and was also over-expressed in some adenomas. IHC staining confirmed the positive relationship between TLR4 staining score in the CRC tumor stroma and epithelium with tumor stage, with up to 47% of colon cancer stroma positive for TLR4 staining. Increased TLR4 expression by IHC was also marginally associated with decreased survival. We now also describe that pericryptal myofibroblasts are responsible for a portion of the TLR4 stromal staining.

Conclusions

Increased TLR4 expression occurs early in colonic neoplasia. TLR4 is associated with the important cancer-related outcomes of survival and stage.

Available only for authorised users

Terzic J, Grivennikov S, Karin E, Karin M: Inflammation and colon cancer. Gastroenterology. 2010, 138 (6): 2101-2114. 10.1053/j.gastro.2010.01.058. e2105CrossRefPubMed

Eckburg PB, Bik EM, Bernstein CN, Purdom E, Dethlefsen L, Sargent M, Gill SR, Nelson KE, Relman DA: Diversity of the human intestinal microbial flora. Science. 2005, 308 (5728): 1635-1638. 10.1126/science.1110591.PubMedCentralCrossRefPubMed

Wells JM, Rossi O, Meijerink M, van Baarlen P: Epithelial crosstalk at the microbiota-mucosal interface. Proc Natl Acad Sci U S A. 2011, 108 (Suppl 1): 4607-4614.PubMedCentralCrossRefPubMed

Poxton IR, Brown R, Sawyerr A, Ferguson A: The mucosal anaerobic gram-negative bacteria of the human colon. Clin Infect Dis. 1997, 25 (Suppl 2): S111-S113.CrossRefPubMed

Zheng L, Riehl TE, Stenson WF: Regulation of colonic epithelial repair in mice by Toll-like receptors and hyaluronic acid. Gastroenterology. 2009, 137 (6): 2041-2051. 10.1053/j.gastro.2009.08.055.PubMedCentralCrossRefPubMed

Ehrchen JM, Sunderkotter C, Foell D, Vogl T, Roth J: The endogenous Toll-like receptor 4 agonist S100A8/S100A9 (calprotectin) as innate amplifier of infection, autoimmunity, and cancer. J Leukoc Biol. 2009, 86 (3): 557-566. 10.1189/jlb.1008647.CrossRefPubMed

Fukata M, Chen A, Vamadevan AS, Cohen J, Breglio K, Krishnareddy S, Hsu D, Xu R, Harpaz N, Dannenberg AJ, Subbaramaiah K, Cooper HS, Itzkowitz SH, Abreu MT: Toll-like receptor-4 promotes the development of colitis-associated colorectal tumors. Gastroenterology. 2007, 133 (6): 1869-1881. 10.1053/j.gastro.2007.09.008.PubMedCentralCrossRefPubMed

Fukata M, Shang L, Santaolalla R, Sotolongo J, Pastorini C, España C, Ungaro R, Harpaz N, Cooper HS, Elson G, Kosco-Vilbois M, Zaias J, Perez MT, Mayer L, Vamadevan AS, Lira SA, Abreu MT: Constitutive activation of epithelial TLR4 augments inflammatory responses to mucosal injury and drives colitis-associated tumorigenesis. Inflamm Bowel Dis. 2011, 17 (7): 1464-1473. 10.1002/ibd.21527.PubMedCentralCrossRefPubMed

Santaolalla R, Sussman DA, Ruiz JR, Davies JM, Pastorini C, España CL, Sotolongo J, Burlingame O, Bejarano PA, Philip S, Ahmed MM, Ko J, Dirisina R, Barrett TA, Shang L, Lira SA, Fukata M, Abreu MT: TLR4 activates the beta-catenin pathway to cause intestinal neoplasia. PLoS ONE. 2013, 8 (5): e63298-10.1371/journal.pone.0063298.PubMedCentralCrossRefPubMed

10.

Gene expression omnibus.http://www.ncbi.nlm.nih.gov/geo/,

11.

Colon cancer tissue microarray case sets.http://dctd.cancer.gov/ProgramPages/cdp/tools_disease-specific.htm,

12.

Wang EL, Qian ZR, Nakasono M, Tanahashi T, Yoshimoto K, Bando Y, Kudo E, Shimada M, Sano T: High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer. Br J Cancer. 2010, 102 (5): 908-915. 10.1038/sj.bjc.6605558.PubMedCentralCrossRefPubMed

13.

Cammarota R, Bertolini V, Pennesi G, Bucci EO, Gottardi O, Garlanda C, Laghi L, Barberis MC, Sessa F, Noonan DM, Albini A: The tumor microenvironment of colorectal cancer: stromal TLR-4 expression as a potential prognostic marker. J Transl Med. 2010, 8: 112-10.1186/1479-5876-8-112.PubMedCentralCrossRefPubMed

14.

RDCTeam: R: A Language and Environment for Statistical Computing. 2010, Vienna, Austria: R Foundation for Statistical Computing

15.

Benjamini Y, Hochberg Y: Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Roy Stat Soc B Met. 1995, 57 (1): 289-300.

16.

Troyanskaya OG, Garber ME, Brown PO, Botstein D, Altman RB: Nonparametric methods for identifying differentially expressed genes in microarray data. Bioinformatics (Oxford, England). 2002, 18 (11): 1454-1461. 10.1093/bioinformatics/18.11.1454.CrossRef

17.

Jorissen RN, Gibbs P, Christie M, Prakash S, Lipton L, Desai J, Kerr D, Aaltonen LA, Arango D, Kruhøffer M, Orntoft TF, Andersen CL, Gruidl M, Kamath VP, Eschrich S, Yeatman TJ, Sieber OM: Metastasis-associated gene expression changes predict poor outcomes in patients with Dukes stage B and C colorectal cancer. Clin Cancer Res. 2009, 15 (24): 7642-7651. 10.1158/1078-0432.CCR-09-1431.PubMedCentralCrossRefPubMed

18.

Sabates-Bellver J, Van der Flier LG, de Palo M, Cattaneo E, Maake C, Rehrauer H, Laczko E, Kurowski MA, Bujnicki JM, Menigatti M, Luz J, Ranalli TV, Gomes V, Pastorelli A, Faggiani R, Anti M, Jiricny J, Clevers H, Marra G: Transcriptome profile of human colorectal adenomas. Mol Cancer Res. 2007, 5 (12): 1263-1275. 10.1158/1541-7786.MCR-07-0267.CrossRefPubMed

19.

Skrzypczak M, Goryca K, Rubel T, Paziewska A, Mikula M, Jarosz D, Pachlewski J, Oledzki J, Ostrowski J: Modeling oncogenic signaling in colon tumors by multidirectional analyses of microarray data directed for maximization of analytical reliability. PLoS ONE. 2010, 5 (10):

20.

Lips EH, van Eijk R, de Graaf EJ, Oosting J, de Miranda NF, Karsten T, van de Velde CJ, Eilers PH, Tollenaar RA, van Wezel T, Morreau H: Integrating chromosomal aberrations and gene expression profiles to dissect rectal tumorigenesis. BMC Cancer. 2008, 8: 314-10.1186/1471-2407-8-314.PubMedCentralCrossRefPubMed

21.

Nishida N, Nagahara M, Sato T, Mimori K, Sudo T, Tanaka F, Shibata K, Ishii H, Sugihara K, Doki Y, Mori M: Microarray analysis of colorectal cancer stromal tissue reveals upregulation of two oncogenic miRNA clusters. Clin Cancer Res. 2012, 18 (11): 3054-3070. 10.1158/1078-0432.CCR-11-1078.CrossRefPubMed

22.

Gray R, Barnwell J, McConkey C, Hills RK, Williams NS, Kerr DJ: Adjuvant chemotherapy versus observation in patients with colorectal cancer: a randomised study. Lancet. 2007, 370 (9604): 2020-2029. 10.1016/S0140-6736(07)61866-2.CrossRefPubMed

23.

André T, Boni C, Mounedji-Boudiaf L, Navarro M, Tabernero J, Hickish T, Topham C, Zaninelli M, Clingan P, Bridgewater J, Tabah-Fisch I, de Gramont A: Oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment for colon cancer. N Engl J Med. 2004, 350 (23): 2343-2351. 10.1056/NEJMoa032709.CrossRefPubMed

24.

Thorsteinsson MKL, Lund LR, Sørensen LT, Gerds TA, Jess P, Olsen J: Gene expression profiles in stage II and III colon cancer. Application of a 128-gene signature. Int J Colorectal Dis. 2012, 27 (12): 1579-1586. 10.1007/s00384-012-1517-4.CrossRefPubMed

25.

Smith JJ, Deane NG, Wu F, Merchant NB, Zhang B, Jiang A, Lu P, Johnson JC, Schmidt C, Bailey CE, Eschrich S, Kis C, Levy S, Washington MK, Heslin MJ, Coffey RJ, Yeatman TJ, Shyr Y, Beauchamp RD: Experimentally derived metastasis gene expression profile predicts recurrence and death in patients with colon cancer. Gastroenterology. 2010, 138 (3): 958-968. 10.1053/j.gastro.2009.11.005.PubMedCentralCrossRefPubMed

26.

Staub E, Groene J, Heinze M, Mennerich D, Roepcke S, Klaman I, Hinzmann B, Castanos-Velez E, Pilarsky C, Mann B, Brümmendorf T, Weber B, Buhr HJ, Rosenthal A: An expression module of WIPF1-coexpressed genes identifies patients with favorable prognosis in three tumor types. J Mol Med (Berl). 2009, 87 (6): 633-644. 10.1007/s00109-009-0467-y.CrossRef

27.

de Sousa E Melo F, Colak S, Buikhuisen J, Koster J, Cameron K, de Jong JH, Tuynman JB, Prasetyanti PR, Fessler E, van den Bergh SP, Rodermond H, Dekker E, van der Loos CM, Pals ST, van de Vijver MJ, Versteeg R, Richel DJ, Vermeulen L, Medema JP: Methylation of cancer-stem-cell-associated Wnt target genes predicts poor prognosis in colorectal cancer patients. Cell Stem Cell. 2011, 9 (5): 476-485. 10.1016/j.stem.2011.10.008.CrossRefPubMed

28.

Reid JF, Gariboldi M, Sokolova V, Capobianco P, Lampis A, Perrone F, Signoroni S, Costa A, Leo E, Pilotti S, Pierotti MA: Integrative approach for prioritizing cancer genes in sporadic colon cancer. Genes Chromosomes Cancer. 2009, 48 (11): 953-962. 10.1002/gcc.20697.CrossRefPubMed

29.

Kaiser S, Park YK, Franklin JL, Halberg RB, Yu M, Jessen WJ, Freudenberg J, Chen X, Haigis K, Jegga AG, Kong S, Sakthivel B, Xu H, Reichling T, Azhar M, Boivin GP, Roberts RB, Bissahoyo AC, Gonzales F, Bloom GC, Eschrich S, Carter SL, Aronow JE, Kleimeyer J, Kleimeyer M, Ramaswamy V, Settle SH, Boone B, Levy S, Graff JM, et al: Transcriptional recapitulation and subversion of embryonic colon development by mouse colon tumor models and human colon cancer. Genome Biol. 2007, 8 (7): R131-10.1186/gb-2007-8-7-r131.PubMedCentralCrossRefPubMed

30.

Gryfe R, Kim H, Hsieh ET, Aronson MD, Holowaty EJ, Bull SB, Redston M, Gallinger S: Tumor microsatellite instability and clinical outcome in young patients with colorectal cancer. N Engl J Med. 2000, 342 (2): 69-77. 10.1056/NEJM200001133420201.CrossRefPubMed

31.

Jorissen RN, Lipton L, Gibbs P, Chapman M, Desai J, Jones IT, Yeatman TJ, East P, Tomlinson IP, Verspaget HW, Aaltonen LA, Kruhøffer M, Orntoft TF, Andersen CL, Sieber OM: DNA copy-number alterations underlie gene expression differences between microsatellite stable and unstable colorectal cancers. Clin Cancer Res. 2008, 14 (24): 8061-8069. 10.1158/1078-0432.CCR-08-1431.PubMedCentralCrossRefPubMed

32.

Earl TM, Nicoud IB, Pierce JM, Wright JP, Majoras NE, Rubin JE, Pierre KP, Gorden DL, Chari RS: Silencing of TLR4 decreases liver tumor burden in a murine model of colorectal metastasis and hepatic steatosis. Ann Surg Oncol. 2009, 16 (4): 1043-1050. 10.1245/s10434-009-0325-8.CrossRefPubMed

33.

Yang H, Zhou H, Feng P, Zhou X, Wen H, Xie X, Shen H, Zhu X: Reduced expression of Toll-like receptor 4 inhibits human breast cancer cells proliferation and inflammatory cytokines secretion. J Exp Clin Cancer Res. 2010, 29: 92-10.1186/1756-9966-29-92.PubMedCentralCrossRefPubMed

34.

Simiantonaki N, Kurzik-Dumke U, Karyofylli G, Jayasinghe C, Michel-Schmidt R, Kirkpatrick CJ: Reduced expression of TLR4 is associated with the metastatic status of human colorectal cancer. Int J Mol Med. 2007, 20 (1): 21-29.PubMed

35.

Adegboyega PA, Mifflin RC, DiMari JF, Saada JI, Powell DW: Immunohistochemical study of myofibroblasts in normal colonic mucosa, hyperplastic polyps, and adenomatous colorectal polyps. Arch Pathol Lab Med. 2002, 126 (7): 829-836.PubMed

36.

Adegboyega PA, Ololade O, Saada J, Mifflin R, di Mari JF, Powell DW: Subepithelial myofibroblasts express cyclooxygenase-2 in colorectal tubular adenomas. Clin Cancer Res. 2004, 10 (17): 5870-5879. 10.1158/1078-0432.CCR-0431-03.CrossRefPubMed

37.

Kalluri R, Zeisberg M: Fibroblasts in cancer. Nat Rev Cancer. 2006, 6 (5): 392-401. 10.1038/nrc1877.CrossRefPubMed

38.

Ban S, Mitsuhashi T, Shimizu M: Immunohistochemical study of myofibroblasts in colorectal epithelial lesions. Arch Pathol Lab Med. 2003, 127 (12): 1551-1553. author reply 1551–1552PubMed

39.

Worthley DL, Giraud AS, Wang TC: Stromal fibroblasts in digestive cancer. Cancer Microenviron. 2010, 3 (1): 117-125. 10.1007/s12307-009-0033-8.PubMedCentralCrossRefPubMed

40.

Otte JM, Rosenberg IM, Podolsky DK: Intestinal myofibroblasts in innate immune responses of the intestine. Gastroenterology. 2003, 124 (7): 1866-1878. 10.1016/S0016-5085(03)00403-7.CrossRefPubMed

41.

University of California Santa Clara genome browser.http://genome.ucsc.edu/cgi-bin/hgTracks?hgHubConnect.destUrl=..%2Fcgi-bin%2FhgTracks&clade=mammal&org=Human&db=hg19&position=chr9%3A120466460-120479766&hgt.suggest=tlr4&hgt.suggestTrack=knownGene&Submit=submit&hgsid=279196009&knownGene=pack

42.

Lynch KW: Consequences of regulated pre-mRNA splicing in the immune system. Nat Rev Immunol. 2004, 4 (12): 931-940. 10.1038/nri1497.CrossRefPubMed

43.

Wells CA, Chalk AM, Forrest A, Taylor D, Waddell N, Schroder K, Himes SR, Faulkner G, Lo S, Kasukawa T, Kawaji H, Kai C, Kawai J, Katayama S, Carninci P, Hayashizaki Y, Hume DA, Grimmond SM: Alternate transcription of the Toll-like receptor signaling cascade. Genome Biol. 2006, 7 (2): R10-10.1186/gb-2006-7-2-r10.PubMedCentralCrossRefPubMed

44.

Grigoryev YA, Kurian SM, Nakorchevskiy AA, Burke JP, Campbell D, Head SR, Deng J, Kantor AB, Yates JR, Salomon DR: Genome-wide analysis of immune activation in human T and B cells reveals distinct classes of alternatively spliced genes. PLoS ONE. 2009, 4 (11): e7906-10.1371/journal.pone.0007906.PubMedCentralCrossRefPubMed

45.

Moon JW, Lee SK, Lee JO, Kim N, Lee YW, Kim SJ, Kang HJ, Kim J, Kim HS, Park SH: Identification of novel hypermethylated genes and demethylating effect of vincristine in colorectal cancer. J Exp Clin Cancer Res. 2014, 33: 4-10.1186/1756-9966-33-4.PubMedCentralCrossRefPubMed

46.

Sheedy FJ, Palsson-McDermott E, Hennessy EJ, Martin C, O’Leary JJ, Ruan Q, Johnson DS, Chen Y, O’Neill LA: Negative regulation of TLR4 via targeting of the proinflammatory tumor suppressor PDCD4 by the microRNA miR-21. Nat Immunol. 2010, 11 (2): 141-147. 10.1038/ni.1828.CrossRefPubMed

Title: In silico and Ex vivo approaches identify a role for toll-like receptor 4 in colorectal cancer
Authors: Daniel A Sussman
Rebeca Santaolalla
Pablo A Bejarano
Monica T Garcia-Buitrago
Maria T Perez
Maria T Abreu
Jennifer Clarke
Publication date: 01-12-2014
Publisher: BioMed Central
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2014
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/1756-9966-33-45

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2014

Blockade of CXCL12/CXCR4 signaling inhibits intrahepatic cholangiocarcinoma progression and metastasis via inactivation of canonical Wnt pathway

KRAS mutations in tumor tissue and plasma by different assays predict survival of patients with metastatic colorectal cancer

Suppression of NAD(P)H-quinone oxidoreductase 1 enhanced the susceptibility of cholangiocarcinoma cells to chemotherapeutic agents

P68 RNA helicase as a molecular target for cancer therapy

The multiple facets of drug resistance: one history, different approaches

Poly (ADP-ribose) polymerase 3 (PARP3), a potential repressor of telomerase activity

Keynote webinar | Spotlight on antibody–drug conjugates in cancer