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Detection of colorectal dysplasia using fluorescently labelled lectins.

Kuo JC, Ibrahim AE, Dawson S, Parashar D, Howat WJ, Guttula K, Miller R, Fearnhead NS, Winton DJ, Neves AA, Brindle KM - Sci Rep (2016)

Bottom Line: Colorectal cancer screening using conventional colonoscopy lacks molecular information and can miss dysplastic lesions.Wheat germ agglutinin (WGA) showed significantly decreased binding to adenomas in the mouse tissue and in sections of human colon from 47 patients.Helix pomatia agglutinin (HGA) distinguished epithelial regions containing NE from regions containing HP, LGD, HGD or C, with 89% sensitivity, 87% specificity and 97% PPV.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Cambridge, UK.

ABSTRACT
Colorectal cancer screening using conventional colonoscopy lacks molecular information and can miss dysplastic lesions. We tested here the ability of fluorescently labelled lectins to distinguish dysplasia from normal tissue when sprayed on to the luminal surface epithelium of freshly resected colon tissue from the Apc(min) mouse and when applied to fixed human colorectal tissue sections. Wheat germ agglutinin (WGA) showed significantly decreased binding to adenomas in the mouse tissue and in sections of human colon from 47 patients. Changes in WGA binding to the human surface epithelium allowed regions containing normal epithelium (NE) or hyperplastic polyps (HP) to be distinguished from regions containing low-grade dysplasia (LGD), high-grade dysplasia (HGD) or carcinoma (C), with 81% sensitivity, 87% specificity and 93% positive predictive value (PPV). Helix pomatia agglutinin (HGA) distinguished epithelial regions containing NE from regions containing HP, LGD, HGD or C, with 89% sensitivity, 87% specificity and 97% PPV. The decreased binding of WGA and HPA to the luminal surface epithelium in human dysplasia suggests that these lectins may enable more sensitive detection of disease in the clinic using fluorescence colonoscopy.

No MeSH data available.


Related in: MedlinePlus

Binding of fluorescently labelled lectins to colorectal luminal surface epithelium.The figure shows a representative example of the binding of WGA conjugated to AF647 to colorectal tissue sections. (a) Luminal surface epithelium was defined as the sole region of interest (ROI), which would be visible at colonoscopy. (b) ROIs of defined length (ca. 500 μm) and thickness (20 μm) were defined at the luminal surface epithelium, using an automated image analysis system (Ariol™), as illustrated by the white dashed-line box. The insets (b,c) are 3.5 × magnifications of the ROIs indicated by the white arrow and triangle in (a), respectively. Scale bars = 250 μm (a) and 70 μm (b,c) μm.
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f2: Binding of fluorescently labelled lectins to colorectal luminal surface epithelium.The figure shows a representative example of the binding of WGA conjugated to AF647 to colorectal tissue sections. (a) Luminal surface epithelium was defined as the sole region of interest (ROI), which would be visible at colonoscopy. (b) ROIs of defined length (ca. 500 μm) and thickness (20 μm) were defined at the luminal surface epithelium, using an automated image analysis system (Ariol™), as illustrated by the white dashed-line box. The insets (b,c) are 3.5 × magnifications of the ROIs indicated by the white arrow and triangle in (a), respectively. Scale bars = 250 μm (a) and 70 μm (b,c) μm.

Mentions: Next, binding of fluorescently-labelled WGA and other lectins to the luminal surface epithelium of fixed human colon tissue sections was investigated (Fig. 2). Lectin binding was compared with histological assessment of paraffin-embedded colorectal samples derived from adenoma lesions collected from 47 patients. ROIs representative of the pathology classes present (Fig. 3a), were analysed (Fig. 3b(i) and Supplementary Fig. 1). Lectin fluorescence signals were averaged to give a score for each class. WGA and HPA binding showed significant differences across the different pathology classes (P < 0.001). WGA showed highest binding to hyperplasia (Fig. 3b(i)) and decreased binding in the progression from normal epithelium to dysplasia (LGD and HGD) and carcinoma (C). A similar trend was observed for HPA binding (Fig. 3c(i)), although HPA bound minimally to hyperplasia. Both lectins showed variable binding to normal epithelium (Supplementary Fig. 2c), which may reflect partial loss of mucus due to FFPE tissue processing. Normal epithelium, immediately adjacent to HGD or C, showed minimal HPA binding but this increased dramatically with distance (>5 mm) (white arrows, row 4, column 4 in Fig. 3a; Supplementary Fig. 2a). WGA binding showed no such differences (row 4, column 3 in Fig. 3a, Supplementary Fig. 2b). Soybean agglutinin (SBA) binding showed a significant decrease with disease progression (P = 0.05), however binding to all classes was relatively weak (Supplementary Fig. 1). Jackfruit lectin (JFL) and peanut agglutinin (PNA) also showed relatively low binding (Supplementary Fig. 1) and no significant trends were observed (P = 0.064 and P = 0.259 for JFL and PNA, respectively).


Detection of colorectal dysplasia using fluorescently labelled lectins.

Kuo JC, Ibrahim AE, Dawson S, Parashar D, Howat WJ, Guttula K, Miller R, Fearnhead NS, Winton DJ, Neves AA, Brindle KM - Sci Rep (2016)

Binding of fluorescently labelled lectins to colorectal luminal surface epithelium.The figure shows a representative example of the binding of WGA conjugated to AF647 to colorectal tissue sections. (a) Luminal surface epithelium was defined as the sole region of interest (ROI), which would be visible at colonoscopy. (b) ROIs of defined length (ca. 500 μm) and thickness (20 μm) were defined at the luminal surface epithelium, using an automated image analysis system (Ariol™), as illustrated by the white dashed-line box. The insets (b,c) are 3.5 × magnifications of the ROIs indicated by the white arrow and triangle in (a), respectively. Scale bars = 250 μm (a) and 70 μm (b,c) μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4829854&req=5

f2: Binding of fluorescently labelled lectins to colorectal luminal surface epithelium.The figure shows a representative example of the binding of WGA conjugated to AF647 to colorectal tissue sections. (a) Luminal surface epithelium was defined as the sole region of interest (ROI), which would be visible at colonoscopy. (b) ROIs of defined length (ca. 500 μm) and thickness (20 μm) were defined at the luminal surface epithelium, using an automated image analysis system (Ariol™), as illustrated by the white dashed-line box. The insets (b,c) are 3.5 × magnifications of the ROIs indicated by the white arrow and triangle in (a), respectively. Scale bars = 250 μm (a) and 70 μm (b,c) μm.
Mentions: Next, binding of fluorescently-labelled WGA and other lectins to the luminal surface epithelium of fixed human colon tissue sections was investigated (Fig. 2). Lectin binding was compared with histological assessment of paraffin-embedded colorectal samples derived from adenoma lesions collected from 47 patients. ROIs representative of the pathology classes present (Fig. 3a), were analysed (Fig. 3b(i) and Supplementary Fig. 1). Lectin fluorescence signals were averaged to give a score for each class. WGA and HPA binding showed significant differences across the different pathology classes (P < 0.001). WGA showed highest binding to hyperplasia (Fig. 3b(i)) and decreased binding in the progression from normal epithelium to dysplasia (LGD and HGD) and carcinoma (C). A similar trend was observed for HPA binding (Fig. 3c(i)), although HPA bound minimally to hyperplasia. Both lectins showed variable binding to normal epithelium (Supplementary Fig. 2c), which may reflect partial loss of mucus due to FFPE tissue processing. Normal epithelium, immediately adjacent to HGD or C, showed minimal HPA binding but this increased dramatically with distance (>5 mm) (white arrows, row 4, column 4 in Fig. 3a; Supplementary Fig. 2a). WGA binding showed no such differences (row 4, column 3 in Fig. 3a, Supplementary Fig. 2b). Soybean agglutinin (SBA) binding showed a significant decrease with disease progression (P = 0.05), however binding to all classes was relatively weak (Supplementary Fig. 1). Jackfruit lectin (JFL) and peanut agglutinin (PNA) also showed relatively low binding (Supplementary Fig. 1) and no significant trends were observed (P = 0.064 and P = 0.259 for JFL and PNA, respectively).

Bottom Line: Colorectal cancer screening using conventional colonoscopy lacks molecular information and can miss dysplastic lesions.Wheat germ agglutinin (WGA) showed significantly decreased binding to adenomas in the mouse tissue and in sections of human colon from 47 patients.Helix pomatia agglutinin (HGA) distinguished epithelial regions containing NE from regions containing HP, LGD, HGD or C, with 89% sensitivity, 87% specificity and 97% PPV.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Cambridge, UK.

ABSTRACT
Colorectal cancer screening using conventional colonoscopy lacks molecular information and can miss dysplastic lesions. We tested here the ability of fluorescently labelled lectins to distinguish dysplasia from normal tissue when sprayed on to the luminal surface epithelium of freshly resected colon tissue from the Apc(min) mouse and when applied to fixed human colorectal tissue sections. Wheat germ agglutinin (WGA) showed significantly decreased binding to adenomas in the mouse tissue and in sections of human colon from 47 patients. Changes in WGA binding to the human surface epithelium allowed regions containing normal epithelium (NE) or hyperplastic polyps (HP) to be distinguished from regions containing low-grade dysplasia (LGD), high-grade dysplasia (HGD) or carcinoma (C), with 81% sensitivity, 87% specificity and 93% positive predictive value (PPV). Helix pomatia agglutinin (HGA) distinguished epithelial regions containing NE from regions containing HP, LGD, HGD or C, with 89% sensitivity, 87% specificity and 97% PPV. The decreased binding of WGA and HPA to the luminal surface epithelium in human dysplasia suggests that these lectins may enable more sensitive detection of disease in the clinic using fluorescence colonoscopy.

No MeSH data available.


Related in: MedlinePlus