Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2017 Jan 24;8(4):7025-7038.
doi: 10.18632/oncotarget.12347.

Mucins and associated O-glycans based immunoprofile for stratification of colorectal polyps: clinical implication for improved colon surveillance

Shiv Ram Krishn  1 , Sukhwinder Kaur  1 , Yuri M Sheinin  2 , Lynette M Smith  3 , Shailendra K Gautam  1 , Asish Patel  4 , Maneesh Jain  1 , Vasthala Juvvigunta  5 , Priya Pai  1 , Audrey J Lazenby  2 , Hemant K Roy  6 , Surinder K Batra  1   2   7
Affiliations

Mucins and associated O-glycans based immunoprofile for stratification of colorectal polyps: clinical implication for improved colon surveillance

Shiv Ram Krishn et al. Oncotarget. .

Abstract

Sessile serrated adenoma/polyps (SSA/P) are premalignant lesions of colorectal cancer that are difficult to distinguish histologically from hyperplastic polyps (HP) of minimal to no malignant potential. Specific markers for differentiating SSA/P from HP can aid clinicians for optimizing colon surveillance intervals. The present study investigates the potential of mucins and associated O-glycans to distinguish SSA/P from HP. Expression of colonic mucins (MUC1, MUC4, MUC17, MUC2, and MUC5AC) and O-glycans [Sialyl LewisA (CA19-9) and Tn/Sialyl-Tn on MUC1] were analyzed in HP (n=33), SSA/P (n=39), and tubular adenoma (TA) (n=36) samples by immunohistochemistry. A significantly reduced expression of MUC4 (p=0.0066), elevated expression of MUC17 (p=0.0002), and MUC5AC (p<0.0001) was observed in SSA/P cases in comparison to HP cases. Interestingly, significantly higher number of SSA/P cases (p<0.0001) exhibited MUC5AC expression in the goblet cells as well as filled the crypt lumen compared to only goblet cells in majority of the HP cases. Improved diagnostic potential was revealed by multivariate logistic regression analysis where combinatorial panel of MUC5AC/MUC17 discriminated SSA/P from HP (SN/SP=85/82%). Finally, the decision tree model based marker panel (CA19-9/MUC17/MUC5AC) predicted HP, SSA/P and TA with SN/SP of 58%/95%, 79%/90% and 97%/83%, respectively. Overall, the mucin and associated O-glycan based panel defined in the present study could aid in discriminating SSA/P from HP to devise better colon surveillance strategies.

Keywords: O-glycans; hyperplastic polyps; mucins; sessile serrated adenoma/polyps; tubular adenomas.

PubMed Disclaimer

Conflict of interest statement

CONFLICTS OF INTEREST

Authors declare no conflict of interest.

Figures

Figure 1
Figure 1. Immunohistochemical expression of mucins and associated O-glycans in colorectal polyps
A. Box plots representing the H-score for mucins (MUC1, MUC4, MUC17, MUC2, and MUC5AC) and O-glycans (CA19-9, Tn/STn-MUC1) expression in HP, SSA/P and TA. A significant downregulation of MUC4 (p = 0.0066), significant upregulation of MUC17 (p = 0.0002) and MUC5AC (p < 0.0001) was observed in SSA/P cases in comparison to HP cases. The box plots further show that compared to SSA/P in TA cases, MUC1 expression was significantly increased (p = 0.0005) while there was a significant decrease in expression of MUC17 (p = 0.0054), MUC2 (p = 0.02), MUC5AC (p < 0.0001) and CA19-9 (p < 0.0001). B. The HP, SSA/P, and TA tissue specimens were probed with MUC1 (mouse monoclonal-HMFG2), MUC4 (mouse monoclonal-8G7), MUC17 (rabbit polyclonal-SN-1139-2), MUC2 (rabbit monoclonal-EPR6145), MUC5AC (mouse monoclonal-45M1), CA19-9 (mouse monoclonal-NS 19.9), and Tn/STn-MUC1 (mouse monoclonal-5E5) antibodies after non-specific blocking with horse serum. All the tissue sections were examined under a microscope, and the expression was evaluated on basis of reddish brown staining by a pathologist. Representative photomicrographs (20X) of Hematoxylin & Eosin staining, mucins (MUC1, MUC4, MUC17, MUC2, and MUC5AC), and O-glycans (Tn/STn-MUC1 and CA19-9) stained tissues of HP, SSA/Ps, and TA were taken. For uniformity, images for all the mucins and O-glycans staining were taken from the same area in a sample of HP, SSA/P and TA respectively. Magnified images in inset shows expression of respective mucins in different cellular compartments. MUC1 expression was significantly high (p = 0.0005) in TA in comparison to HP and SSA/P. Strong expression of MUC4 was observed in adjacent normal colon. Expression of MUC4 was significantly reduced in SSA/P (p = 0.0066) compared with HP. Significantly high immunoreactivity of MUC17 was observed in SSA/P lesions compared to HP and TA. Compared to HP and SSA/P, MUC2 expression was significantly reduced in TA (p = 0.0018, and 0.020 respectively). MUC5AC expression was significantly high in HP and SSA/P compared to TA (p < 0.0001). HP, SSA/P, and TA expressed Tn/STn-MUC1 at similar levels. CA19-9 expression was significantly high in HP (p < 0.0001) and SSA/P (p < 0.0001) compared to TA. NC: adjacent normal colon; HP: hyperplastic polyps; SSA/P: sessile serrated adenoma/polyps; TA: tubular adenoma.
Figure 2
Figure 2. Differential localization of mucins in colorectal polyp subtypes
Differential localization pattern for MUC5AC and MUC4 were observed for polyp subtypes. MUC5AC expression was absent in normal colon, restricted to the goblet cells (GC) in HP, and present in the goblet cells and lumen of colon crypts in SSA/P. In case of TA, the MUC5AC expression was observed in goblet cells but at lower intensity. In normal colon, MUC4 expression was observed throughout the colon crypts (CC) while it was absent on luminal surface epithelium (LE). In the HP and TA, MUC4 expression was observed throughout CC including luminal epithelium. In contrast to this, in 30% SSA/P cases, MUC4 expression was limited to lower one-third of colon crypts. The differential localization pattern of MUC4 and MUC5AC could provide useful molecular adjunct to histological classification of colorectal polyps. CC: colon crypts; GC: goblet cells; LE: luminal surface epithelium; HP: hyperplastic polyps; SSA/P: sessile serrated adenoma/polyps; TA: tubular adenoma.
Figure 3
Figure 3. Combined performance of mucins and associated O-glycans for discriminating colorectal polyp subtypes
The combined utility of significantly altered mucins and O-glycans as markers for differential identification of polyp subtypes was evaluated utilizing multivariate regression models in conjunction with ROC curve based analysis. A. ROC curves representing diagnostic potential of combinatorial panels of mucins and associated O-glycans for differentiating SSA/P from HP. MUC17+MUC5AC combination differentiated SSA/P from HP with the AUC of 0.87. The model has 83% accuracy (60/72) in predicting SSA/P vs. HP. B. MUC1 and MUC17+CA19-9 combination differentiated SSA/P from TA with the AUC of 0.99. the model has 95% accuracy (71/75) in predicting SSA/P vs. TA. C. Decision tree based model to accurately define the combined diagnostic efficacy of mucins and associated O-glycans for polyp stratification. The panel of three markers CA19-9/MUC17/MUC5AC differentiated precancerous polyps (SSA/P and TA) from benign polyps (HP). Polyp sections having H-Score < 0.525 for CA19-9 predicts the presence of TA. For tissues sections having H-Score ≥ 0.525 for CA19-9, and H-score ≥ 0.75 for transmembrane mucin MUC17 the lesion is likely to be SSA/P. In cases where MUC17 H-score is < 0.75, and MUC5AC H-score is ≥ 1.05, then SSA/P is predicted and if MUC5AC H-score < 1.05, then HP is predicted. The tree based model has 79% accuracy (85/108) in predicting all 3 groups simultaneously.HP: hyperplastic polyps; SSA/P: sessile serrated adenoma/polyps; TA: tubular adenoma.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Jain P, Mondal SK, Sinha SK, Mukhopadhyay M, Chakraborty I. Diagnostic and prognostic significance of different mucin expression, preoperative CEA, and CA-125 in colorectal carcinoma: A clinicopathological study. J Nat Sci Biol Med. 2014;5:404–408. - PMC - PubMed
    1. Carethers JM, Jung BH. Genetics and Genetic Biomarkers in Sporadic Colorectal Cancer. Gastroenterology. 2015;149:1177–1190. - PMC - PubMed
    1. Okugawa Y, Grady WM, Goel A. Epigenetic Alterations in Colorectal Cancer: Emerging Biomarkers. Gastroenterology. 2015;149:1204–1225. - PMC - PubMed
    1. Pancione M, Remo A, Colantuoni V. Genetic and epigenetic events generate multiple pathways in colorectal cancer progression. Patholog Res Int. 2012;2012:509348. - PMC - PubMed
    1. Grady WM, Carethers JM. Genomic and epigenetic instability in colorectal cancer pathogenesis. Gastroenterology. 2008;135:1079–1099. - PMC - PubMed

Substances