Nomenclature for Histologic Assessment of Intestinal Tumors in the Rodent

Nomenclature for Histologic Assessment of Intestinal Tumors in the Rodent

Hyperplasia:gross thickening of the mucosa; some growths may be pedunculated; mitosis are always located in the lower two thirds of the mucosa; nuclei lack significant atypia, are basally located, ovoid to round, usually uniformly dark, with occasional visible nucleoli ; crypts take on a star-shaped appearance in tangential sections; herniation (pseudoinvasion) of epithelium through the muscularis mucosae may occur. Hyperplasia can be further categorized as diffuse, focal, multifocal, associated with inflammation (type and severity noted), and also can be graded as to severity
Aberrant crypt foci (ACF):microscopically in whole-mount colonic tissue that usually is stained with methylene blue; one or more crypts larger than most crypts in the field, have a thickened layer of epithelial cells that stain more intensely with methylene blue, often have a slit-shaped luminal opening, have an increased pericryptal space, and are elevated from the focal plane of the microscope (ACF are not a histologic diagnosis).
Gastrointestinal Intraepithelial neoplasia (GIN):histologically apparent areas of dysplasia that are not visible grossly (>0.5-1.0 mm); in human and veterinary pathology, these lesions may be referred to as microadenoma, microcarcinoma, carcinoma in situ, and focal areas of dysplasia; lesions may involve single or multiple glands; is compatible with dysplastic aberrant crypt foci (ACF) that previously have been identified in the same unembedded tissue
Adenoma:benign, circumscribed neoplasm composed of tubular and/or villous structures lined by dysplastic epithelium; herniation (pseudoinvasion) through the muscularis mucosae may occur; categorized by the following criteria:
  1. macroscopic growth pattern:sessile, broad-based or pedunculated (polypoid)-attached by a stalk
  2. histologic type:tubular - at least 75% of adenoma composed of branching tubules in lamina propria (usually pedunculated); villous - at least 75% of adenoma composed of leaf- or finger-like processes of lamina propria covered by epithelium (usually sessile; also known as papillary); tubulovillous - adenoma composed of 25-75% of both tubular and villous structures (usually pedunculated; also known as papillotubular)
  3. grade of dysplasia:based on the most severely dysplastic area of each tumor      low grade: branching or elongation of crypts with some reduction of interglandular stroma; low N/C ratio; cell nuclei elongate, crowded, appear stratified, with regular nuclear membranes, fine chromatin, and inconspicuous nucleoli; nuclei maintain polarity with respect to the basement membrane; mucus secretion usually present
         high grade: exhibits both architectural and cytologic changes; marked reduction of interglandular stroma with complex irregularity of glands with cribriform (sieve-like) structures and back-to-back glands; high N/C ratio; cell nuclei large, ovoid to round, with loss of polarity with respect to the basement membrane; cytologic atypia is more pronounced with marked, irregular nuclear membranes, with aberrant chromatin pattern not basally located (cleared, vesicular, clumped, or densely hyperchromatic chromatin); large, conspicuous nucleoli; mucus secretion usually absent; numerous mitoses with abnormal mitotic figures
Herniation:glands have penetrated through the muscularis mucosae (see Table 3).
Adenocarcinoma:malignant neoplasia of glandular epithelium composed of tubular and/or villous structures penetrating through the muscularis mucosa; categorized by the following criteria:
  1. grade of differentiation:well differentiated, moderately differentiated, or poorly differentiated;
  2. 2. histologic type:tubular/tubulovillous/villous adenocarcinoma; mucinous adenocarcinoma (>50% of tumor composed of extracellular mucin, signet-ring cells can be present); signet-ring cell adenocarcinoma (>50% of tumor composed of signet-ring cells); undifferentiated carcinoma (no glandular structure to differentiate; can be uniform or pleomorphic)


Summary: Different GEM models of intestinal cancer display characteristic disease manifestations. For example, lesions appear primarily in the cecum and colon in some mouse lines and in the small intestine in others. Gross morphology is also influenced by genotype; pedunculated polyps carry Apc mutations (regardless of the initial mutation) while sessile lesions do not. Disease severity also differs among the GEM models. For example, theApcmodels and some of the MMR GEM have a high incidence of adenoma but only rarely develop carcinomas. Others, such as the Smad3 and Tgfβ1xRag2 mutants exhibit adenoma and carcinomas with equal frequency. In these models carcinomas are observed more frequently in the large intestine than in the small intestine.

Mouse models of intestinal cancer exhibit some important differences with respect to human disease. In general, adenomas are the most common lesion in murine intestinal cancer models. However, unlike humans, adenomas occur throughout the intestine in mice, with no predilection for the large bowel. Another important difference is that metastasis is rare in mice. Only one mouse line,Smad3-/-(83), exhibits metastasis of colonic adenocarcinomas to lymph nodes and liver, which is a common occurrence in human colorectal carcinomas. There is a critical need for a consistent model of metastasis.

Gut flora is an important modifier of intestinal tumor development. For example, when derived in a germ-free environment, Il-10-/-, IL-2-/-, and Tgfβ1-/-XRag2-/- mice do not develop colonic lesions (77,91,94). Despite evidence of its importance, the role of the gut flora is often overlooked in studies involving GEM. Understanding the role of bacterial flora in the intestine is necessary to understand disease initiation and progression in murine models of intestinal tumor formation. The precise role of bacteria in the development of lesions is not understood, but is an important issue that should be addressed in each model with an unexpected or invasive phenotype.

An important issue in interpreting the phenotypes of GEM is accurate discrimination between invasive cancer and herniation. In the mouse, the muscularis mucosa is very thin, thus, the mucosa is able to penetrate the muscle easily. Herniation can occur through weak points where blood vessels and lymphatics traverse the muscle. Herniation can also result from pressure exerted by the polyp that can push basal crypt cells through the muscularis mucosa. Both of these mechanisms are enhanced by inflammatory conditions.

The Consensus report on the intestinal pathology of GEM (58) includes a classification system (link to box: Features that distinguish?) to facilitate consistent and accurate distinction between carcinoma and mucosal herniation. At least one of the following features should be present to classify a lesion as a carcinoma: desmoplasia, loss of mucosal lining in invading glands, and the presence of irregular, sharp or angulated glands. Particular caution is necessary when evaluating inflamed areas because of the severe fibrosis and frequent disruption and herniation of glandular tissue that occurs in many chronic inflammatory bowel disease models.

Herniation of the intestinal mucosa of an Msh3-/- mouse. The invading cells include multiple cellular types, and there is no desmoplastic response in the herniated intestinal lesion. There is severe desmoplasia and loss of mucosa in the carcinoma.
Herniation of the intestinal mucosa of an Msh3-/- mouse. The invading cells include multiple cellular types, and there is no desmoplastic response in the herniated intestinal lesion. There is severe desmoplasia and loss of mucosa in the carcinoma.



Features that Distinguish Invasive Carcinoma versus Herniation
  1. (Reprinted with permission from reference 58)


(Reprinted with permission:
Boivin GP, Washington K, Yang K, Ward JM, Pretlow TP, Russell R, Besselsen DG, Godfrey VL, Doetschman T, Dove WF, Pitot HC, Halberg RB, Itzkowitz SH, Groden J, Coffey RJ
Gastrointestinal Pathology of mouse models of intestinal cancer: consensus report and recommendations.
Gastroenterology. 2003 Mar;124(3):762-77.
PMID: 12612914)