Lgr5 is Widely Expressed in the Human Gut and

MASSACHUSETTS
GENERAL HOSPITAL
PATHOLOGY
Lgr5 is Widely Expressed in the Human Gut and Overexpression is a
HARVARD
Biomarker of Colonic and Barrett's-related Dysplasia
MEDICAL SCHOOL
Jacob R Bledsoe MD, Mohammad Shahid MD, David T. Ting MD, Miguel N. Rivera MD, Ömer H Yilmaz MD, PhD,
Vikram Deshpande MD
Department of Pathology, Massachusetts General Hospital, Boston, MA
Background
• Leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5) is a robust marker of
intestinal stem cells and has been implicated as a cancer stem cell marker.1-2
• Much of the data on the normal distribution of Lgr5 expression in humans is extrapolated
from mouse studies, and few studies have examined Lgr5 expression in human neoplasia.1
•Branch chain nucleic acid in-situ hybridization is a highly sensitive assay which can detect
as few as one to two copies of a target messenger RNA sequence per cell.3
•To better define the distribution of Lgr5 expression in humans, we applied this assay to a
wide range normal and neoplastic human tissues.
Methods
Figure 2 Lgr5 expression at the GEJ
Compared to low level expression in the non-goblet cell columnar-lined esophagus and normal gastric cardiac
mucosa (A; mean 2.2 dots), an increase in Lgr5 expression was seen in BE with intestinal metaplasia (B; mean 89 ;
p<0.001). Lgr5 expression in BE with dysplasia was significantly higher than in BE without dysplasia a (C; mean
375;p=0.003).
• Evaluation of Lgr5 expression was performed on tissue microarrays of 13 non-neoplastic
gastrointestinal tissues from 65 people, as well as 37 cases of Barrett's esophagus (BE) with
and without dysplasia, 12 colonic tubular adenomas, 10 sessile serrated adenomas, and 20
cases of pancreatic neoplasia.
• Lgr5 and GAPDH (house keeping gene) expression was evaluated using a highly sensitive
in situ hybridization (ISH) assay based on branched DNA technology (QuantiGene®
ViewRNA, Affymetrix, Santa Clara, CA).
• Quantitative analysis was performed by counting distinct Lgr5 mRNA foci (red dots) within
the 200x200 uM field of each tissue core with highest expression.
Figure 3 Lgr5 expression in the pancreas
Lgr5 was not expressed in the normal pancreas (A; inset: GAPDH control shows preserved RNA), but is expressed in
pancreatic neoplasia: intraductal papillary mucinous neoplasia (B; inset: high power) and pancreatic ductal
adenocarcinoma (C; inset: H&E).
Lgr5 distribution in normal human
gastrointestinal tissues
Figure 1 Lgr5 expression in the colon
Compared to the corresponding normal colon, where Lgr5 expression in normal colonic
crypts is limited to the base of crypts (A) with a mean 100 red dots per 200x200uM field, an
expanded Lgr5+ cell population and increased Lgr5 expression was seen in colonic tubular
adenomas (B; mean 2317 red dots; p<0.001) and colitis-associated dysplasia, but not
sessile serrated adenomas.
Table 1 Lgr5
Tissue type
Lgr5+ distribution
expression in normal
Esophagus (squamous) Absent
human tissues
Lgr5 expression in nonGastric cardia
Base of gastric pit
neoplastic human
Gastric antrum
Mid gastric pit (+/- base)
gastrointestinal and
Gastric body/fundus
Rare positive cells,
hepatobiliary tissues was
isthmus/neck region
similar to prior murine
Small intestine
Base of crypts
findings, except that no
expression was seen in
Ampulla
Absent
the normal liver or
Colon
Base of crypts
pancreas. Large
Appendix
Base of crypts
extrahepatic bile ducts
include the common bile
Anus
Absent
duct and main pancreatic
Liver
Absent
duct.
Gallbladder
Base of mucosal folds
Large extrahepatic bile
ducts
Peribiliary glands
(deep>superficial)
Pancreas
Absent
Conclusions
• Lgr5+ cells occupy a predictable niche in the human
gastrointestinal tract.
• Neoplastic colonic, esophageal, and pancreatic lesions
consistently overexpressed Lgr5 with a stepwise
increase in expression along the metaplasia-dysplasia
sequence.
• These results have implications for the diagnosis and
surveillance of gastrointestinal neoplasia, Lgr5-targeted
chemoprevention in neoplasia, and for the use of Lgr5+
cells in regenerative medicine.
References:
1. Barker N, Tan S, Clevers H. Lgr proteins in epithelial stem cell biology. Development 2013;140:2484 2494.
2. Barker N, van Es JH, Kuipers J, et al. Identification of stem cells in small intestine and colon by marker gene
Lgr5. Nature 2007;449:1003–1007.
3. Player AN, Shen LP, Kenny D, et al. Single-copy gene detection using branched DNA (bDNA) in situ
hybridization. J Histochem Cytochem 2001;49:603-612.

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