Fig. 6.1.
Radial EUS showing the five circumferential layers of the normal rectal wall. (Courtesy of Drs. Vikesh K. Singh and Vivek Kumbhari, The Johns Hopkins Hospital, Maryland, USA)
ERUS-FNA, developed in the early 1990s, enhances the diagnostic yield of ERUS by providing material for microscopic evaluation. The diagnostic yield of ERUS-FNA has been reported to be 89–96 %, whereas, the success rate for imaging investigation alone without histologic findings was 81.8 %. The overall sensitivity, specificity, and positive and negative predictive values of ERUS-FNA are reported to be 89 % (74–100 %), 79 % (50–100 %), 89 % (74–100 %), and 79 % (51–100 %), respectively. FNA especially improves the accuracy of ERUS in the diagnosis of recurrent rectal cancer.
Rapid on-site evaluation of the aspirated material by a cytopathologist or cytotechnologist for specimen adequacy increases the diagnostic yield. Specimens obtained by ERUS-FNA can easily be processed for cytologic diagnosis and for ancillary tests including, but not limited to, immunostains, cultures, flow cytometry, or molecular studies.
ERUS-FNA Procedure
ERUS- FNA is usually well tolerated, with conscious sedation providing increased comfort. The procedure can be performed by a gastroenterologist or radiologist in either the endoscopy or the ultrasound unit on an outpatient basis. ERUS is preferably performed on a clean, empty rectum. Laxative enemas are usually sufficient; however standard colonoscopy preparation optimizes imaging. A rigid probe or a flexible echoendoscope with a radial transducer is used for ultrasound examination of intraluminal rectal lesions. A linear transducer is routinely used when FNA sampling is needed. The bowel wall is seen under ERUS as five alternating hyper- and hypoechoic layers. Informed consent is obtained after the risks and benefits of the procedure are explained to the patient.
The patient is placed in the left lateral decubitus position. Administered medications, oxygenation, and vital signs are monitored constantly during the procedure. A colonoscopy is performed for the direct visualization of the mucosa. Subsequently, ERUS with the use of a high-frequency radial-imaging echoendoscope is done. Water is used as necessary to provide an adequate acoustic interface. After the lesion is detected, measurements are documented and ERUS-FNA is performed as described for EUS-FNA of the upper digestive tract in Chap. 2. Color Doppler ultrasound is used for identifying and avoiding any vessels.
The Role of the Cytopathologist or Cytotechnologist
A cytopathologist or cytotechnologist evaluates the aspirated material on-site. The number of passes and specimen triage are determined by cytology findings on-site. In most cases of solid masses, three or more passes are performed. Needle rinses and clotted blood are transferred to the transport medium or formalin as needed for making cell blocks and possible further ancillary tests. Flow-cytometry studies are done on aspirated material when the cytology findings are suspicious for lymphoma on immediate interpretation. Material is sent to the microbiology laboratory for cultures when the cytology findings indicate an infectious process. In the case of benign cystic lesions or abscess, on- site evaluation allows the endoscopist to aspirate the fluid and drain the cyst or abscess.
Indications
ERUS-FNA is a safe and efficient method used in clinical practice for evaluation of any suspicious intramural, extramural, and or perirectal mass lesions by providing cytology material for microscopic examination. ERUS-FNA is used for evaluation of (1) the depth of transmural invasion in colorectal cancer; (2) tumor staging of colorectal, anal, and other locoregional cancers; (3) tumor invasion of perirectal fat; (4) nodal status and nodal metastasis; (5) suspected recurrent cancers in and adjacent to surgical anastomoses; (6) tumor recurrence or metastasis; (7) the presence of malignancy before chemotherapy, radiation therapy, and/or aggressive surgery; (8) pelvic masses; (9) intramural and submucosal masses; and (10) perirectal cystic masses.
Contraindications
Contraindications include the inability to clearly visualize a lesion or a tumor mass, or presence of a vessel interposed in the path between the needle and the target.
Complications
The complication rate is 1–2 %, which is similar to that of computed tomography (CT) or percutaneous ultrasound-guided FNA. The major reported complications are infections in cystic lesions, and bleeding. Most reported complications are from EUS-FNA of upper gastrointestinal tract or pancreas. Bacteremia occurs after ERUS-FNA at a rate similar to that of colonoscopy, but it does not warrant the prophylactic administration of antibiotics. Occasionally, prophylactic antibiotics are prescribed, depending upon the patient’s general condition. The risk of tumor seeding along the biopsy tract is very small, with only rare anecdotal cases reported.
The Role of ERUS-FNA in Colorectal Cancer Staging
The prognosis of rectal cancer correlates with pathologic staging at the time of diagnosis. ERUS has become the method of choice for locoregional colorectal cancer staging.
ERUS is the most accurate modality for assessing the local depth of invasion into the rectal-wall layers (T staging in the TNM classification); the prefix ‘‘u’’ is suggested in T staging by use of an ultrasound. ERUS can detect early cancers, residual carcinoma in the rectal wall, and local recurrence at an anastomosis site.
For nodal (N) staging, involvement is usually suspected if a lymph node is round, hypoechoic, and > 5 mm in diameter, features which may differentiate it from a non-neoplastic node. ERUS alone is not as accurate for predicting nodal metastasis as it is for tumor depth. Preoperative ERUS-FNA may enhance the identification of extra-mesenteric lymph node metastasis outside a standard radiation field or mesorectal resection margin, and it may affect the choice of surgical or chemoradiation treatment strategy.
Accurate preoperative staging determines both the type of surgery performed and the decision to use neoadjuvant therapy. Inaccurate staging tends to result from over-staging because of associated (1) peri-tumoral inflammation; (2) difficulty distinguishing malignant from reactive lymphadenopathy; (3) chemoradiation effects such as inflammation, edema, necrosis, or fibrosis; and (4) postradiation changes such as fibrosis that is difficult to distinguish from residual tumor. Preservation of the anal sphincter is the goal in treatment of patients who have low-lying, locally advanced rectal cancer, because it decreases morbidity and improves the quality of life. Preoperative chemoradiation in these patients improves local control and is associated with an increased rate of sphincter preservation. Complete tumor staging is compromised in approximately 14 % of patients because of luminal tumor stenosis.
Comparative studies have demonstrated that the accuracy of ERUS T and N colorectal cancer staging is superior to that of CT and equivalent to that of MRI. Overall, ERUS accuracy in numerous studies ranges from 80 to 95 % for T-staging and from 70 to 75 % for N-staging. These levels are slightly higher than the 75–85 % for T staging and 60–70 % for N-staging for MRI and 65–75 % and 55–65 %, respectively, for helical CT.
A succinct description of the most common intra- and extramural masses of the colorectum evaluated and diagnosed by ERUS and ERUS-FNA are covered in the next sections.
ERUS-FNA of Benign Intramural Colorectal Masses
Leiomyoma
Leiomyoma of the colorectum is a rare benign tumor arising from the muscularis layer of the colorectal wall of adults, predominantly males with a median age of 60 years. It presents as a well-delineated, white, and firm polypoid nodule. A conservative excision is therapeutic.
Histopathology
The tumor is a well-circumscribed submucosal mass consisting of benign appearing smooth-muscle cells. Necrosis and atypical mitoses are not features of leiomyoma, whereas nuclear pleomorphism can be seen.
Immuno-Profile
The tumor cells are immunoreactive for smooth muscle actin and desmin and non-reactive for CD117, S-100 protein, and CD34.
FNA Findings
Aspirated material of ERUS-FNA is usually scant. It consists of benign-appearing, spindled smooth-muscle cells arranged mainly in tissue fragments (Fig. 6.2). Necrosis, atypical mitoses, or nuclear pleomorphism is not seen. Diagnostic immunohistochemistry can be done on the cell blocks.
Fig. 6.2.
Leiomyoma. Tissue fragment showing cohesive spindle cells with ill-defined cytoplasmic borders, elongated nuclei with blunt ends, and a dense, glassy stroma (DiffQuik stain, X400)
ERUS Features
A well-defined, homogeneous hypoechoic intramural mass, often originating from the muscularis propria, is identified.
Lipoma
Lipoma of the large bowel is a rare, benign, often single, and submucosal adipose-tissue tumor. Occasionally, lipomas may present as multiple yellowish polyps. Lipoma is seen in adults and affects both genders. Large lipomas can cause constipation, abdominal pain, rectal bleeding, and intussusception.
Histopathology
Lipoma is an encapsulated mass composed of mature adipose tissue exhibiting small, uniform nuclei. Focal fat necrosis may be seen.
Immuno-Profile
Lipomas are immunoreactive for vimentin and S-100 protein.
Molecular Profile
One half of lipomas show aberrations in the chromosome 12q13–15 segment. The target gene for rearrangement is HMGA2 (HMGA2/LLP and HMGA2/RDC1).
FNA Findings
Aspirated smears show small fragments of mature adipocytes and grossly visible fat vacuoles (Fig. 6.3). The adipocytes contain a large, single lipid vacuole and a small nucleus located at the periphery.
Fig. 6.3.
Lipoma. ERUS-FNA shows a fragment of mature adipose tissue with large single vacuoles pushing the small nuclei to the periphery (DiffQuik stain, X200)
ERUS Features
Lipoma is a well-defined, often hyperechogenic submucosal mass (Figs. 6.4 and 6.5).
Fig. 6.4.
Lipoma. Linear EUS showing a large iso- and hyperechoic pedunculated polypoid mass that probably originated from the submucosal layer (calipers). The muscularis propria is intact (hypoechoic band inferior to the lower caliper). Retrosigmoid resection revealed a 3.5 cm submucosal lipoma. (Courtesy of Drs. Vikesh K. Singh and Joanna Law, The Johns Hopkins Hospital, Maryland, USA)
Fig. 6.5.
ERUS-FNA of a submucosal mass, with the sampling needle inside the mass. (Courtesy of Drs. Vikesh K. Singh and Joanna Law, The Johns Hopkins Hospital, Maryland, USA)
Schwannoma
Schwannoma is a rare, benign colorectal tumor of adults and affects both genders equally. The tumor is usually a well-circumscribed submucosal mass.
Histopathology
Schwannoma is unencapsulated and is surrounded by lymphocytes. Intralesional lymphocytes can also be seen. The tumor pattern is spindle- or plexiform. The spindle cells are arranged in interlacing bundles; however, in contrast with peripheral Schwannomas or GISTs, a distinct palisading is not identified. The plexiform pattern shows nodules of spindle cells. Focal nuclear atypia or rare mitotic figures are occasionally seen.
Immuno-Profile
The cells are immunoreactive for S-100 protein and GFAP and nonreactive for c-Kit, smooth-muscle actin, and desmin; CD34 immunoreactivity may be focal and weak.
Molecular Profile
Gastrointestinal Schwannomas are not associated with c-Kit or with neurofibromatosis 2 (NF2) gene mutations.
FNA Findings
On cytology, the smear shows spindle cells arranged singly or in tissue fragments. The nuclei are oval or wavy. Scattered small lymphocytes, an important clue for the diagnosis, are always present and must be thoroughly searched (Fig. 6.6).
Fig. 6.6.
Schwannoma. Loosely cohesive tissue fragment with irregular borders. Round to oval to spindle-shaped nuclei with tapered or pointed ends embedded in a fibrillary stroma. Notice the sprinkled small lymphocytes (Papanicolaou stain, X400)
ERUS Features
The mass is relatively well-defined, hypoechogenic, and submucosal. Features are indistinguishable from those of GIST.
Heterotopic Pancreas
Ectopic pancreatic tissue is a relatively frequent congenital anomaly. It is most common in the upper gastrointestinal tract, including the stomach, duodenum, and jejunum, and less common in the lower GI tract, particularly the colon.
Gross examination reveals a well-circumscribed, lobulated, yellow tan, firm nodule. Central umbilication is seen in cases located in the submucosa. Histologic examination shows normal pancreatic acini and ducts. Islet cells are seen in less than half of the cases. Ectopic pancreas is immunoreactive for amylase, lipase, and trypsinogen. No genetic abnormality is detected. Cytology smears show clusters of benign pancreatic acini and ductal epithelium (Fig. 6.7).
Fig. 6.7.
Benign acinar cells with uniform round, eccentrically placed nuclei and granular cytoplasm in an ectopic pancreas (DiffQuik stain, X400. Courtesy of Dr. Yener Erozan, The Johns Hopkins Hospital, Maryland, USA)
Gastric Heterotopia
Gastric heterotopia is a rare condition that may occur in the anus and rectum. It is slightly more common in males than in females, with a mean age of 21 years. Patients complain of rectal pain, bleeding, or irritable bowel syndrome. In asymptomatic patients gastric heterotopia can be identified incidentally on cancer screening. The lesion may appear as a polyp, diverticulum, or ulcer on colonoscopy. A fundic type gastric mucosa is the most common histologic finding. Cytology smears show benign columnar, parietal, and oxyntic cells.
ERUS-FNA of Malignant Intramural Colorectal Masses
Colorectal Adenocarcinoma
Colorectal carcinoma is the most common carcinoma of the gastrointestinal tract. It is the third most common cancer in the United States, affecting males and females equally at a mean age of 60 years. It is the second leading cancer-related cause of death. The pathogenesis of colorectal carcinoma is related to personal, familial, and environmental risk factors. Individuals with a history of inflammatory bowel disease (ulcerative colitis, Crohn’s disease), adenomas (tubular, villous, or tubulovillous), sessile serrated polyps, prior colorectal cancer, and/or those who have a first-degree relative with invasive colorectal cancer are at high risk of developing colorectal cancer. Inherited syndromes, such as Lynch syndrome (also known as hereditary nonpolyposis colorectal cancer), polyposis syndromes including classical familial adenomatous polyposis, attenuated familial adenomatous polyposis, MUTYH-associated polyposis (MAP), juvenile polyposis syndrome, Peutz-Jeghers syndrome, serrated polyposis syndrome, Cowden syndrome, and Li-Fraumeni syndrome also are associated with a high risk of colorectal cancer. Diets high in red meat or processed meat and low in fiber, as well as smoking, alcohol consumption, a high body mass index, and a low level of physical activity are considered personal lifestyle risk factors. Colorectal cancer screening, recommended at age of 50 for men and women with average risk, has decreased the mortality rate due to colorectal cancer because of early detection of precancerous lesions. Colorectal cancer most commonly occurs in the cecum, ascending colon, sigmoid colon, and rarely in the anal canal (where it must be distinguished from extension from a rectal primary). The tumor is either exophytic or infiltrating.
Histopathology
The well- to moderately-differentiated colorectal carcinoma consists of elongated, pseudostratified columnar cells forming glands of various sizes. The cells exhibit a high nuclear-to-cytoplasmic ratio, hyperchromatic nuclei with prominent nucleoli, and moderate amounts of finely vacuolated cytoplasm. Variable mitotic activity, foci of necrosis, acute and chronic inflammatory response, and pools of excess mucin are common findings. Tumor invasion promotes a desmoplastic stromal reaction Poorly differentiated carcinoma grows in solid sheets with no distinct glandular formation. Focal squamous differentiation or neuroendocrine differentiation can be seen. Other histologic variants of colorectal carcinoma are mucinous, signet-ring-cell, and, less commonly, clear cell, anaplastic, hepatoid, basaloid (cloacogenic), and medullary.
Immuno-Profile
Colorectal adenocarcinoma is immunoreactive for CK20, CDX2, CEA, MUC1, and MUC3. CK7 is negative; however, it can be expressed in rectal adenocarcinoma. The tumor cells are nonimmunoreactive with DPC4 (SMAD4), HepPar1, and MUC2.
Molecular Profile
RAS mutations including both KRAS and NRAS should be investigated in all patients who have metastatic colorectal adenocarcinoma; at the very least, exon 2 KRAS mutation status should be determined. KRAS and NRAS mutations are similar in both primary and/or metastatic colorectal adenocarcinoma. BRAF V600E mutation is associated with a poor prognosis. Testing for NRAS, KRAS, and BRAF V600E mutations is performed on formalin-fixed, paraffin-embedded tissue; PCR-based amplification and DNA sequencing are the recommended methods. Microsatellite instability is associated mainly with mucinous adenocarcinoma or poorly differentiated carcinoma.
FNA Findings
The cytomorphologic findings are similar to the histologic findings. The malignant cells are arranged as single or in clusters or as tissue fragments. Glandular or acinar formation is seen in well- or moderately-differentiated adenocarcinoma. The cells are tall and columnar, with a high nuclear-to-cytoplasmic ratio. The nuclei are elongated and hyperchromatic, with prominent nucleoli and atypical mitotic figures. The cells contain moderate and finely vacuolar cytoplasm. Necrosis and inflammatory cells are commonly prominent in the background (Fig. 6.8).
Fig. 6.8.
Rectal adenocarcinoma. The smear shows a complex flat aggregate of malignant cells with large nuclei, irregular nuclear membranes, and prominent nucleoli. Pseudo-glandular lumens are present (Papanicolaou stain, X400)
ERUS Features
The mass is heterogeneous and hypoechoic, originates in the mucosa, and shows variable invasion into the wall, with irregular infiltrating margins. ERUS accurately shows the depth of tumor invasion (Fig. 6.9) and evaluates regional lymph nodes, if they are larger than 2–3 mm. It is difficult to evaluate the primary site of the tumor in the mucosal layer when the tumor is ulcerated.
Fig. 6.9.
Rectal adenocarcinoma. Transrectal radial EUS shows tumor invasion through the submucosa (arrow) into deep layers. (Courtesy of Drs. Vikesh K. Singh and Joanna Law, The Johns Hopkins Hospital, Maryland, USA).
Squamous Cell Carcinoma
Primary colonic squamous carcinoma is exceedingly rare. Risk factors include mechanical trauma, local radiotherapy, and chronic schistosomiasis. In the absence of risk factors or a history of a distant squamous primary, colonic squamous carcinoma is a diagnosis of exclusion. Squamous cell carcinoma of the anus (15 % of anal cancers) and the anal canal (80 % of anal cancers) is seen in adults and mainly affects females in the sixth and seventh decades of life. In the last 20 years, there has been a shift in the age of diagnosis, with a higher incidence in younger individuals, more commonly in those with immunosuppression, particularly HIV. Rectal bleeding, a nodular or ulcerated mass, and pain are common clinical findings. The tumor can invade deeply in perirectal soft tissue, both proximally and distally. The treatment of choice includes combined neoadjuvant chemoradiation therapy, which achieves a remarkably good response, followed by radical surgery (abdominoperineal resection) only for persistent or locally recurrent tumors.
Histopathology
The tumors vary in their differentiation, keratinization, and subtypes. Keratinized squamous cell carcinoma is characterized by intercellular bridges and keratin pearls. Large-cell, non-keratinizing squamous cell carcinoma lacks keratin pearls and may present diagnostic challenges. Subtypes of squamous cell carcinoma include verrucous carcinoma, carcinoma with mucinous microcysts (mucoepidermoid), and small-cell (anaplastic) carcinoma, the latter two with a poor prognosis. Immunostains differentiate small-cell anaplastic carcinoma from small-cell undifferentiated neuroendocrine carcinoma.