tumori neuroendocrine

NANETS Consensus Guidelines for the Diagnosisof Neuroendocrine Tumor

Aaron I. Vinik, MD, PhD,* Eugene A. Woltering, MD, Richard R. P. Warner, MD,Martyn Caplin, MD, Thomas M. ODorisio, MD,|| Gregory A. Wiseman, MD,

Domenico Coppola, MD,# and Vay Liang W. Go, MD**

Neuroendocrine tumors (NETs) are rare, slow-growing neo-plasms characterized by their ability to store and secretedifferent peptides and neuroamines.1 Some of these substancescause specic clinical syndromes,2 whereas other may have ele-vated plasma or urine levels that are not associated with specicsyndromes or symptom complexes Unfortunately, there is noBideal neuroendocrine tumor marker,[3 but according to the pre-sentation, the sensitivity and specicity of each marker vary, andit is generally possible to choose those of greatest value for eachclinical syndrome.

The biochemical markers are those hormones or aminessecreted by the neuroendocrine cells from which these tumorsare derived. Some of these are not specic to any tumor, but areproduced and secreted by most NETs, whereas other biochem-ical markers are more specic to the type of tumor and wheretheir quantication can lead to the suspicion or conrmation ofthe presence of such a tumor.

The annual incidence of NETs has risen to 40 to 50 casesper million, perhaps because of better diagnosis and the avail-ability of highly specic and sensitive ways to measure thesetumors products, improved immunohistochemistry, and enhancedtechniques for tumor detection. Thus, the perceived increase inincidence may not be a real change in the incidence of the disease.

There are a number of impediments to the diagnosis ofthese tumors. They are rare, comprising less than 2% of gas-trointestinal (GI) malignancies, and are therefore not high onthe list of causes of specic symptom complexes. Symptomsthemselves are often nonspecic and do not lend themselvesreadily to identifying the specic underlying tumor. In addition,the manifestations are protean and mimic a variety of disorders.Tumors may be found incidentally on laparoscopy for abdom-inal pain or during the surgical removal of an appendix or evenduring a computerized tomographic scan of the abdomen forunexplained symptoms. Lung carcinoids may present with he-moptysis or asthma-like symptoms, and midgut carcinoids maybe confused with irritable bowel syndrome (IBS). The naturalhistory of this disease is invariably attended by a long history ofvague abdominal symptoms, a series of visits to a primary carepractitioner, and referral to a gastroenterologist, often with amisdiagnosis of IBS. These symptoms persist with a median

latency to correct diagnosis of 9.2 years by which time the tu-mor has metastasized, causing symptoms such as ushing anddiarrhea and progressing on its slow but relentless course untilthe patient dies. Clearly, a greater index of suspicion and a car-cinoid tumor prole screen are warranted for all patients pre-senting with Btraditional IBS symptoms.[Midgut carcinoids areassociated with mesenteric brosis, which can compress mes-enteric vessels and cause bowel ischemia and malabsorption,which may be found in the absence of an abdominal mass. Thediagnosis of metastases to the liver is generally more obvious butoften still takes place only after a delay of many years. Eventhen, an incorrect diagnosis is not uncommon. Unless biopsymaterial is examined for the secretory peptides chromogranin,synaptophysin, or neuron-specic enolase (NSE), tumors maybe labeled erroneously as adenocarcinoma, with a negative im-pact on physicians attitudes regarding management and under-estimation of prospects for survival.4 The common symptomaticmanifestations of patients with carcinoid tumors are illustratedin Tables 1 and 2.

FlushingIt is vitally important to obtain a good history of the nature

of the ushing to ascertain whether this can be ascribed to a NET.Is the ushing wet or dry? If dry, it is almost always due to aNET. If it is wet, it may still be due to a NET, but the diaphoresisis due to coexistent anxiety. Intermittent ushing suggests men-opause or a NET, for example, systemic mastocytosis or pheo-chromocytoma. Constant ushing is found with alcoholism,polycythemia, andmitral valve disease, either stenosis or prolapse.If there is a facial rash, it may be rosacea, and if more diffuse,consider mastocytosis with the attendant dermatographia or der-matomyositis. If there are associated symptoms or signs such asdiarrhea, this suggests a NET, either carcinoid or medullary car-cinoma of the thyroid (MCT). Headache occurs with mastocytosisor pheochromocytoma, and syncope suggests autonomic neu-ropathy, pheochromocytoma, or epilepsy. Flushing in NETs isquite different depending on the location of the tumor.

ForegutThe ushing in foregut carcinoid tumors is dry, long lasting,

and intense and has a purplish or violet hue in contrast to thecommon red/pink hue seen in other NE-related ushing. It isrelated to telangiectasia and skin hypertrophy mostly in the faceand upper neck but can also involve the limbs, and it can lead to aleonine appearance after repeated episodes.

MidgutIn midgut tumors, it is faint pink to red and involves the face

and upper trunk as far as the nipple line. The ush is initiallyprovoked by exercise, alcohol, and food-containing tyramines(eg, blue cheese, chocolate, red sausage, and red wine). Withtime, the ush may occur spontaneously and without provoca-tion. It usually is ephemeral, lasting only a few minutes, and may

NANETS GUIDELINES

Pancreas & Volume 39, Number 6, August 2010 www.pancreasjournal.com 713

From the *Eastern Virginia Medical School, Strelitz Diabetes Center forEndocrine and Metabolic Disorders, Norfolk, VA; Louisiana State Univer-sity Health Sciences Center, New Orleans, LA; Mt Sinai School of Medi-cine, New York, NY; Royal Free Hospital London, London, UK; ||HoldenComprehensive Cancer Center University of Iowa, Iowa City, IA; MayoClinic, Rochester, MN; #H. Lee Moffitt Cancer Center University of SouthFlorida, Tampa, FL; and **David Geffen School of Medicine, UCLA LosAngeles, CA.Reprints: Aaron Vinik, MD, PhD, Eastern Virginia Medical School Strelitz

Diabetes Research Center, 855 W Brambleton Ave, Norfolk, VA 3510(e-mail: [email protected]).

Copyright * 2010 by Lippincott Williams & Wilkins

Copyright 2010 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.

occur many times per day; however, over many years, patientsmay develop a persistent ush with a purpuric malar and nasalcomplexion.

The differential diagnosis of ushing includes the post-menopausal state, simultaneous ingestion of chlorpropamideand alcohol, panic attacks, MCT, autoimmune epilepsy, auto-nomic neuropathy, and mastocytosis.5 To differentiate all thosecauses from a carcinoid tumor, besides knowing the differencesin the characteristics of the ushing, it is also necessary to knowwhat is producing the ushing (Table 3).

Flushing in carcinoid syndrome has been ascribed toprostaglandins, kinins, and serotonin (5-HT). With the advent ofsophisticated radioimmunoassay methods and region-specicantisera, a number of neurohumors now are thought to be se-creted by carcinoid tumors, including 5-HT, dopamine, hista-mine, and 5-hydroxyindoleacetic acid (5-HIAA), kallikrein,

substance P, neurotensin, motilin, somatostatin, vasoactive in-testinal polypeptide (VIP), prostaglandins, neuropeptide K, andgastrin-releasing peptide.

Several provocative tests have been developed to identifythe cause of ushing in carcinoid syndrome. These tests arebased on the need to distinguish the ushing from that found in ahost of other conditions particularly in panic syndrome in whichthe associate anxiety and phobias usually establish the cause, butfrequently, the physician and patient need reassurance that thereis no underlying malignancy.

Feldman and ODorisio6 have previously reported the inci-dence of elevated levels of plasma neuropeptide concentrationsin patients with ushing. Despite the elevated basal concentrationsof substance P and neurotensin, these authors were able to doc-ument further increases in these neuropeptides during ethanol-induced facial ushing.

TABLE 1. Clinical Presentations, Syndromes, Tumor Types, Sites, and Hormones4

Clinical Presentation Syndrome Tumor Type Sites Hormones

Flushing Carcinoid Carcinoid Midgut/foregut, adrenalmedulla, gastric

Serotonin, CGRP, calcitonin

Medullary carcinomaof thyroid

C cell tumor Metanephrine andnormetanephrine

pheochromocytoma Tumor of chromaffin cells Thyroid C cellsAdrenal and sympatheticnervous system

Diarrhea abdominalpain and dyspepsia

Carcinoid, WDHHAsyndrome, ZES,PP, MCT

Carcinoid, VIPoma,gastrinoma, PPoma,medullary carcinomathyroid, mastocytoma

As above, pancreas,mast cells, thyroid

As above, VIP, gastrin,PP, calcitonin

Diarrhea/steatorrhea Somatostatin,bleeding GI tract

Somatostatinoma,neurofibromatosis

Pancreas, duodenum Somatostatin

Wheezing Carcinoid Carcinoid Gut/pancreas/lung SP, CGRP, 5-HTUlcer/dyspepsia ZES Gastrinoma Pancreas/duodenum GastrinHypoglycemia Whipple triad Insulinoma, sarcoma,

hepatomaPancreas, retroperitonealliver

Insulin, IGF-1, IGF-11

Dermatitis Sweet syndrome,pellagra

Glucagonoma Pancreas Glucagon

Carcinoid Midgut SerotoninDementia Sweet syndrome Glucagonoma Pancreas GlucagonDiabetes Glucagonoma Glucagonoma Pancreas Glucagon

Somatostatin Somatostatinoma Pancreas SomatostatinDeep vein thrombosis,steatorrhea, cholelithiasis

Somatostatin Somatostatinoma Pancreas Somatostatin

neurofibromatosis DuodenumSilent, livermetastasis

Silent PPoma Pancreas PP

This table summarizes the suggested approach to diagnose a NET based on the clinical presentation, the tumor type, their sites of origin, and thepossible means of diagnosis and the biochemical markers that should be measured.

CGRP indicates calcitonin gene-related peptide; WDHHA, watery diarrhea, hypokalemia, hyperchlorhydria, and acidosi.

TABLE 2. Clinical Presentation, Syndrome, Tumor Type of Ectopic Pancreatic Tumors, and the Hormones Produced

Clinical Presentation Syndrome Tumor Type Sites Hormones

Acromegaly Acromegaly, Gigantism NET Pancreas islet GHRHCushing Cushing NET Pancreas islet CRH, ACTHPigmentation Pigmentation NET Pancreas islet Melanocyte-stimulating hormoneAnorexia, nausea, vomiting, abdominal pain Hypercalcemia NET Pancreas islet PTH-rP

Vinik et al Pancreas & Volume 39, Number 6, August 2010

714 www.pancreasjournal.com * 2010 Lippincott Williams & Wilkins


Ahlman and colleagues7 reported the results of pentagastrin(PG) provocation in 16 patients with midgut carcinoid tumorsand hepatic metastases. All patients tested had elevated urinary5-HIAA levels, and 12 had profuse diarrhea requiring medica-tion. Pentagastrin uniformly induced facial ushing and GIsymptoms in patients with liver metastases, but it had no effectin healthy control patients. All patients with PG-induced GIsymptoms demonstrated elevated 5-HT levels in peripheralblood. Administration of a 5-HT-receptor antagonist had no ef-fect on 5-HT release but completely aborted the GI symptoms.The authors emphasized the improved reliability of PG com-pared with calcium infusion, another provocative test popu-larized by Kaplan and colleagues,8 and pointed out that PGprovocation occasionally can be falsely negative in patients withsubclinical disease. Vinik et al9 reported that PG uniformly in-duced ushing in patients with gastric carcinoid tumors that wasassociated with a rise in circulating levels of SP in 80%. Thus,Substance P is a neurohumor that may be involved in the ushingof carcinoid syndrome.

Substance P has also been found in tumor extracts andplasma from patients with carcinoid tumors and, in 1 reportedcase, was useful for tumor localization. Neurokinin A (NKA),its amino-terminally extended form, neuropeptide K, and SPare a group of peptides (ie, tachykinins) with common biologicproperties. Norheim and colleagues measured peptide responsesto PG or ingestion of food or alcohol in 16 patients with meta-static carcinoid tumors and demonstrated 2-fold or greater in-creases in NKA and neuropeptide K in 75% of patients, as wellas variable increases in SP in approximately 20% of patients.

Conlon and colleagues10 used region-specic antisera to SPand NKA to measure circulating tachykinins during a meal-induced ush in 10 patients with metastatic carcinoid tumors.Five patients had undetectable levels of NKA and SP after stim-ulation, thus suggesting that elevated tachykinin concentrationsare not a constant feature of such patients. The authors alsostudied the effect of a somatostatin-analog administration onmeal-induced tachykinin responses in 3 patients with carcinoidtumors. Flushing was aborted in 2 patients, but tachykinin levelswere only partially suppressed, indicating that these peptides

cannot be solely responsible for the carcinoid ush. When thediagnosis of the underlying cause of ushing has been established,pathogenesis-oriented treatment can be very helpful.5

Janet et al also performed a study, in which they usedpatients with metastasizing ileocecal 5-HTYproducing carcinoidtumors and looked for the relationship of ushing to tachykininproduction. They concluded that metastasizing ileocecal 5-HTYproducing carcinoids producemany biologically active substanceswith partially overlapping biological functions. The biologicalprocesses underlying the specic symptoms of the carcinoidsyndrome are probably multifactorial. They conrmed resultsfrom earlier studies showing that tachykinins and 5-HIAA levelsare elevated in patients with daily episodes of ushing. The hor-mone effects were not mutually independent. It is possible thatthe development of ushing is the result of multihormonal stim-ulation. Other biologically active substances, such as kallikrein,and prostaglandins, may also contribute.11

Diarrhea, Abdominal Pain, DyspepsiaThe key question to ask is whether the diarrhea persists with

fasting. Diarrhea of NETs is always secretory, whereas diarrheafrom other GI causes is usually malabsorptive. Perhaps the mostvexing question is to differentiate the diarrhea and abdominalpain and dyspepsia from IBS. Irritable bowel syndrome affects alarge proportion of the American population (6%Y12%), and inPakistan, Mexico, and Brazil, it may be as high as 40%. Ab-dominal pain and discomfort are the most prominent symptomsprompting consultation with a physician. Irritable bowel syn-drome occurs more frequently in young white females, begin-ning before the age of 35 years in 50% of cases, and thesymptoms typically do not disturb sleep, quite unlike that ofNETs. Bleeding, fever, weight loss, and persistent severe painare not features of IBS. Most sufferers (7.4%) report alternatingconstipation and diarrhea distinguishing them from NETs, andthe diarrhea does not persist with fasting. Secretory diarrhea ischaracteristic of NETs, causing large-volume stools, and persistswith fasting, and there is no osmotic gap between serum andstool. There are several causes of secretory diarrhea that need tobe taken into consideration in the differential diagnosis: waterydiarrhea, hypokalemia, hyperchlorhydria, and acidosis syndrome,the Zollinger-Ellison syndrome (ZES), carcinoid tumors, MCT,secreting villous adenoma of the rectum, surreptitious laxativeabuse, and idiopathic. Diarrhea that disappears with the use of aproton pump inhibitor (PPI) is very suggestive of a gastrinomabecause acid overproduction precipitates bile salts; inactivates li-pase, amylase, and trypsin; and damages the intestinal mucosa.Another confounding issue is the change in the character of thediarrhea from a secretory diarrhea to a malabsorptive one with theuse of somatostatin analogs. The stools become foul smelling;there is an inability to ush the toilet; and the stools oat on thesurface of the water and contain undigested food particles becausesomatostatin analogs cause inhibition of pancreatic enzyme se-cretion and the intestinal absorption of uid and nutrients.

Neuroendocrine tumors can produce diarrhea by differentmechanisms, depending on their secretory products. Gastrin canincrease the acid secretion by the stomach, which in turn inac-tivates lipase, amylase, and trypsin and damages the mucosa ofthe small bowel, leading to decreased absorption and impaireddigestion in the small bowel, exceeding the absorptive capacityof the colon, what gives an increased fecal volume and mal-absorptive syndromes and sometimes steatorrhea. On the otherhand, carcinoid or other NETs can produce other substancessuch as VIP, pancreatic polypeptide (PP), SP, calcitonin gene-related peptide, and/or thyrocalcitonin, all of which will acton the small bowel increasing the secretion of uids and ions,

TABLE 3. Differential Diagnosis of Flushing andRecommended Tests

Clinical Condition Tests

Carcinoid Serotonin, 5-HIAA, NKA, TCP, PP,CGRP, VIP, SP, PGD2, PGE1, PGF2

MCT Calcitonin, Ca2+ infusion, RETprotooncogene

Pheochromocytoma CgA, plasma free metanephrines,urine metanephrines, VMA,epinephrine, norepinephrine,glucagon stimulation, MIBG

Diabetic autonomicneuropathy

HRV, 2hs PP glucose

Menopause FSHEpilepsy EEGPanic attack Pentagastrin, ACTHMastocytosis Plasma histamine, urine tryptaseHypomastia and mitralvalve prolapse

Cardiac echocardiogram

EEG indicates electroencephalography; FSH, follicle stimulatinghormone; HRV, heart rate variability; PGD, prostaglandin D2; PGE,prostaglandin E.

Pancreas & Volume 39, Number 6, August 2010 Neuroendocrine Tumor, Biochemical Test, Diagnosis

* 2010 Lippincott Williams & Wilkins www.pancreasjournal.com 715


which in turn will also exceed the colonic absorptive capacity,producing an increased fecal volume as well as a great lossesof potassium and bicarbonate.

A disturbing cause of diarrhea that may be very difcultto detect is laxative abuse, and, in all circumstances, a KOH stoolpreparation to detect laxatives is mandatory. Measurement of in-testinal secretion by passing a multilumen tube and quantifyingelectrolytes and water transport, in addition to the measurementof stool electrolytes, which should account for the total osmolar-ity, will help to exclude laxative abuse but is rarely performed.

It is important to mention that Janet et al found, in their studyof tachykinins and NETs, that there is an association betweenthe elevation of tachykinins and the severity of the diarrhea. Theyconcluded that all biochemical markers concentrations were ele-vated in patients with daily episodes of diarrhea, although theassociation between increased plasma tachykinins and the severityof diarrhea was independent of both chromogranin A (CgA) and5-HIAA concentrations.11

BronchoconstrictionWheezing due to bronchospasm occurs in up to one third of

patients with carcinoid syndrome. Lung function tests show aprolonged forced expiratory volume in the rst second. Differ-ential diagnoses are asthma and chronic obstructive pulmonarydisease. In the carcinoid syndrome, the cause of bronchocon-striction is usually substance P, histamine, or 5-HT that should bemeasured in patients who present with this symptom.4

Dyspepsia or Peptic UlcerThe ZES is characterized by peptic ulcers and diarrhea that

responds to therapy with PPIs, in the setting of hypergastrinemiaand low gastric pH. Gastrinomas are localized 90% of the time inthe Bgastrinoma triangle.[ As discussed in the previous section,the measurements that should be drawn for these tumors arefasting serum gastrin (FSG) and gastric acid output and, to dif-ferentiate gastrinoma from other cause of hypergastrinemia, asecretin- or calcium-stimulated gastrin.

HypoglycemiaThe Whipple triad (symptoms of hypoglycemia, low blood

glucose levels G40 mg/dL, and relief of symptoms with glucose)is the clinical presentation of insulinomas, but other causesshould be ruled out. Patients with noninsulinoma pancreato-genous hypoglycemia syndrome present with postprandial neu-roglycopenia symptoms (within 4 hours of meal ingestion) andhave negative 72 hours fasting test and negative tumor locali-zation studies, and on histological diagnosis, hypertrophy ornesidioblastosis rather than an insulinoma is found.12,13 Otherpossible causes that should be thought of are fasting, autoim-mune (insulin antibodies), counter-regulatory hormone de-ciency, drug-induced and factitious hypoglycemia. To excludeall the other causes, clinical suspicion together with measure-ment of hormones or peptides should be used.

In the case of hypoglycemia, the recommended biochem-ical markers are insulin, insulin-like growth factor 2 (IGF-2),C-peptide, glucagon-like peptide type 1 (GLP-1), glucose depen-dent insulin releasing peptide, sulfonylurea, adrenocorticotropichormone (ACTH), growth hormone (GH), insulin antibodies, andliver enzymes.4

Dumping SyndromeThis manifestation occurs after surgery when the pylorus

has been resected or inactivated. It can be early, when symptomsresemble shock, or late, which presents as hypoglycemia. For the

diagnosis of this syndrome, a provocative test is done, giving thepatient a high-calorie, carbohydrate-rich breakfast with 750 kcal(21 g protein, 30 g fat, and 99 g carbohydrate) that shouldbe ingested in 10 minutes to produce the maximum response.After completion of the meal, blood sample is collected at 10,15, 30, 45, 60, 120, and 180 minutes; to measure glucose, in-sulin, C-peptide, motilin, PP, and GLP-1 levels.4 An exaggeratedinsulin and GLP-1 response to the meal is found in gastric by-pass patients with the syndrome, although the case and rela-tionship between the hormonal overproduction and the clinicalsyndrome remain controversial.

PellagraPellagra is caused by the deciency of niacin due to the

detour of the tryptophan pathway toward the production of in-creased amounts of 5-HT.

DIAGNOSIS OF GASTRIC CARCINOID TUMORSFasting serum gastrin levels are important to differentiate

types 1 and 2 gastric carcinoids from type 3. Gastrin levels areelevated in both types 1 and 2 gastric carcinoids but not in type 3. Itis important to note that patient with type 1 gastric carcinoid arehypochlorhydric or achlorhydric, whereas patients with type 2gastric carcinoids have high acid levels. 5-Hydroxyindoleaceticacid levels are generally not useful in patients with gastric carci-noid tumors, because development of the carcinoid syndrome isuncommon.14 Furthermore, carcinoid syndrome, if it occurs inthese patients, is reported to be characteristically atypical withnormal 5-HT and 5-HIAA levels, although a recent study reportsthat the typical carcinoid syndrome can occur in rare patients withgastric carcinoids.1 Plasma CgA levels are recommended becauseCgA is frequently elevated in both patients with types 1 and 2 andtype 3 gastric carcinoid tumors,16 and changes in CgA levels maybe helpful in follow-up.16 Chromogranin A should be used withcaution as a marker of disease activity in patients treated withsomatostatin analogs, because these agents signicantly reduceplasma CgA levels, a change that may be more reective ofchanges in hormonal synthesis and release from tumor cells thanan actual reduction in tumor mass.16,17 In patients on stable dosesof somatostatin analogs, consistent increases in plasma CgA levelsover time may reect loss of secretory control and/or tumorgrowth. Plasma CgA levels have also been shown to have prog-nostic value in patients with metastatic disease.16,18

PATHOLOGYNeuroendocrine tumors arise throughout the body and

share certain basic characteristics. It is suggested that all tumorsare classied according to site, differentiation (well vs poorlydifferentiated); a marker of cell proliferation, for example, Ki-67;grade and stage; and hormones or amines produced and markersof behavior such as CgA and synaptophysin. Tumor differentia-tion refers to the extent of resemblance to the normal cellularcounterpart or loss of this resemblance. Thus, NETs may be wellor poorly differentiated. Tumor grade refers to the degree of bio-logical aggressiveness; this may be related to differentiation butnot necessarily so. Tumor stage refers to the extent of spread ofthe tumor. The extent of invasion into the organ of origin andinvolvement of nodes or distant sites are critical factors. Thereare a number of different systems to classify, grade, and stageNETs, and although the criteria differ among systems, the un-derlying basic data are similar. A review of nomenclature, grad-ing, and staging system has been summarized in previous articleson the pathological classication of NETs adapted by the devel-opment of the NANETS Consensus Guidelines.




IMMUNOHISTOCHEMISTRYAlthough most agree that a mitotic rate or Ki-67 is neces-

sary in specic cases, whether Ki-67 staining is needed in allcases remains hotly debated. An experienced pathologist fa-miliar with NETs will likely be able to determine the tumorsgrade in most resected specimens, and a Ki-67 can be obtainedas needed in difcult cases. In small biopsy specimens, theremay not be sufcient material to differentiate between grade1 versus 2 neuroendocrine carcinomas with or without Ki-67This marker can be helpful when there is large amount of crushartifact in grading these tumors.

SPECIAL PATHOLOGICAL CONSIDERATIONAMONG NETS OF THE MIDGUT

In addition to classic NETs, mixed histology tumors havingneuroendocrine as well as glandular features, such as goblet cellcarcinoids and adenocarcinoids, can be observed. Among mid-gut tumors, these are more frequent in the appendix and cecum.For example, mixed histology tumors account for 1% of jejunal/ileal NETs and 7.3% of cecal NETs (P G 0.001).19 Although thenumber of fraction of appendiceal NETs having mixed histologytumors in the Surveillance, Epidemiology, and End Resultsregistry is even higher, the exact percentage is difcult to as-certain. This is due to the fact that small appendiceal carcinoidsare often considered benign and not reported to the registries,and mixed histology tumors such as goblet cell carcinoids aregenerally considered malignant and more likely to be reported.

Similarly, the distribution of poorly differentiated NETsamong midgut tumor is different. Poorly differentiated NETsaccount for only 0.9% and 1.1% of appendiceal and jejunal/ilealNETs, respectively. They, however, account for 14.2% of NETsarising from the cecum.19

These pathological descriptors provide important informa-tion for patient management as mixed histology tumors such asgoblet cell can be more aggressive and prone to peritoneal dis-semination. Poorly differentiated NETs are often rapidly pro-gressive and require cytotoxic chemotherapy (see separate article).

MOLECULAR GENETICSThe genetics of neuroendocrine tumorigenesis have yet to

be elucidated. Although small familial clusters of midgut car-cinoids have been described, there are no known genetic cancersyndromes associated with the development of midgut NETs.Tumors have clustered in several small families without multipleendocrine neoplasia type I (MEN I), and multiplicity of tumors isa feature on a quarter of isolated cases. Among sporadic midgutcarcinoids, several studies examining the genetics of ileal car-cinoid tumors using comparative genomic hybridization or mi-crosatellite markers have shown frequent allelic deletion ofchromosome 18.20,21 On an epigenetic level, midgut NETs havebeen found to have global hypomethylation.22 There are few dataabout genetic aspects in NETs of the appendix or cecum. Tumormultiplicity is much less frequent in the appendix and cecumthan the ileum.

BLOOD AND URINE BIOMARKERSIN MIDGUT NETS

Several circulating tumor markers have been evaluated forthe follow-up management of NETs. Although these can be veryuseful for follow-up, isolated elevation of marker levels is gen-erally not sufcient for diagnosis without tissue conrmation.The most important of these markers, CgA, is a 49-kd acidic

polypeptide that is widely present in the secretory granules ofneuroendocrine cells. Plasma CgA is elevated in 60% to 100% ofpatients with either functioning or nonfunctioning NETs. Thesensitivities and specicities of CgA for the detection of NETsrange between 70% and 100%.23Y26 Chromogranin A level maycorrelate with tumor volume, but care should be taken in mea-suring CgA and interpreting the results. For example, becausesomatostatin analogs are known to affect blood levels of CgA,serial CgA levels should be measured at approximately the sameinterval from injection in patients receiving long-acting somato-statin analogs. Spuriously elevated levels of CgA have also beenreported in patients using PPIs, in patients with renal or liverfailure, and in those patients with chronic gastritis.

Urinary 5-HIAA (24-hour collection) is a useful laboratorymarker for carcinoid tumors. It is a surrogate measure of 5-HTmetabolism that tightly linked to the presence of carcinoidsyndrome. It is also perhaps more useful than the direct mea-surement of 5-HT as serum 5-HT varies considerably during theday according to activity and stress level. The specicity of thistest has been reported to be 88%.27 However, certain foods andmedications can increase urinary 5-HIAA levels and should beavoided during specimen collection.28 High 5-HT concentra-tions occur with the ingestion of bananas, kiwis, pineapple,plantains, plums, and tomatoes. Moderate elevations are foundwith avocado, black olives, spinach, broccoli, cauliower, egg-plant, cantaloupe, dates, gs, grapefruit, and honeydew melon.Drugs that can increase 5-HIAA are acetanilide, phenacetin,reserpine, glyceryl guaiacolate (found in many cough syrups),and methocarbamol. Drugs that can decrease 5-HIAA levelsinclude chlorpromazine, heparin, imipramine, isoniazid, levo-dopa, monoamine oxidase inhibitors, methenamine, methyldo-pa, phenothiazines, promethazine, and tricyclic antidepressants.Thus, there are a variety of confounding factors in the mea-surement of 5-HT and its metabolite, 5-HIAA, in addition to thefact that foregut carcinoids do not produce 5-HIAA but ratheronly 5HTP, limiting the usefulness of these measures as a di-agnostic or screening tool.

Another useful blood marker, NSE, is a dimer of the gly-colytic enzyme enolase. Neuron-specic enolase is present in thecytoplasmic compartment of the cell, and its serum level isthought to be unrelated to the secretory activity of the tumor.26

Although it is less specic than CgA, it may be a useful markerfor follow-up of patients with a known diagnosis of NETs.

A variety of other secreted molecules can be measuredamong patients with midgut NETs. These include other chro-mogranins such as chromogranins B and C, pancreastatin, andsubstance B. The general principle of biomarker measurement isto identify a few biomarkers that are elevated in the particularpatient in question and follow these over time. It is generally notnecessary to check every biomarker at every visit.

PANCREATIC NETSPancreatic NETs (PNETs) have an estimated incidence of

less than 1 per 100,000 individuals.29Y31 Pancreatic NETs aredivided into 2 groups: those associated with a functional syn-drome due to ectopic secretion of a biologically active substanceand those that are not associated with a functional syndrome(generally called nonfunctional PNETs [NF-PNETs]).29Y32 Thisdistinction is important for clinical presentation, diagnosis, andtreatment of these tumors. Functional PNETs include insulino-mas, gastrinomas, VIPomas, somatostatinomas, glucagonomas,GH-releasing factorYsecreting (GRFomas), and a group of lesscommon PNETs including PNETs secreting ACTH (ACTHomas)and causing Cushing syndrome, PNETs causing the carcinoid




syndrome, PNETs causing hypercalcemia, and very rarely PNETsectopically secreting luteinizing hormone, rennin, or erythropoi-etin.29 Functional PNETs and NF-PNETs also frequently secretea number of other substances (chromogranins, NSE, subunits ofhuman chorionic gonadotropin, neurotensin, ghrelin), but theydo not cause a specic hormonal syndrome.29Y32 In terms ofrelative frequency, NF-PNETs are at present the most frequentlyfound PNET, occurring approximately twice as frequently asinsulinomas, which are generally more frequent than gastrinomas,following by glucagonomas 9 VIPomas 9 somatostatinomas 9others.29,30,32,33

Pancreatic NETs can occur both sporadically and in patientswith various inherited disorder.25,30 Pancreatic NETs occur in80% to 100% of patients with MEN I; in 10% to 17% of patientswith von Hippel-Lindau syndrome (VHL); up to 10% of patientswith von Recklinghausen disease (neurobromatosis type 1[NF-1]), and occasionally in patients with tuberous sclerosis.34

Each of these is an autosomal dominant disorder.34 Of these dis-orders, MEN I is the most frequent, in patients with PNETs.34,35

Multiple endocrine neoplasia type I is caused by mutations inchromosome 11q13 region resulting in alterations in the MEN Igene, which encodes for a 610Yamino acid nuclear protein,menin, which has important effects on transcriptional regulation,genomic stability, cell division, and cell cycle control.34 Patientswith MEN I develop hyperplasia or tumors of multiple endo-crine and nonendocrine tissues including parathyroid adenomas(95%Y100%) resulting in hyperparathyroidism, pituitary adeno-mas in 54% to 65%, adrenal adenomas (27%Y36%), variouscarcinoid tumors (gastric, lung, thymic) (0%Y10%), thyroid ade-nomas (up to 10%), various skin tumors (80%Y95%), centralnervous system tumors (up to 8%), and smooth muscle tumors(up to 10%).34 In MEN I patients, 80% to 100% develop pan-creatic NF-PNETs, but in most patients, they are small and mi-croscopic, causing symptoms in only 0% to 13%.34 Gastrinomas(980% duodenal) develop in 54% of MEN I patients; insulin-omas develop in 18%, and glucagonomas, VIPomas, GRFomas,somatostatinomas in less than 5%.34 In VHL, 98% of all thePNETs that develop in 10% to 17% of the patients are NF-PNETs, in 0% to 10% of NF-1 patients developing a PNET, theyare characteristically duodenal somatostatinomas, which do notcause the somatostatinoma syndrome, and in tuberous sclerosis,rare functional and NF-PNETs are reported.34

INSULINOMAInsulinoma patients characteristically present with symp-

toms of hypoglycemia, especially neuroglycopenic symp-toms (confusion, altered consciousness) and symptoms due tosympathetic overdrive (weakness, sweating), which are usuallymade worse by fasting.29,36,37 The diagnosis of insulinomacan be established by determining plasma proinsulin, insulin, C-peptide, and glucose levels, which are usually performed duringa 72-hour fast.25,32,33 It is important to realize that insulin levelsare increasingly being determined by immunochemiluminescentassays or specic immunoradiometric assays that do not cross-react with proinsulin and give lower values than that obtainedwith most insulin radioimmunoassays, which can effect theproposed criteria listed in many reviews for diagnosis, whichwere based on radioimmunoassay results.29

There are 6 criteria for the diagnosis of insulinomas: docu-mented blood glucose levels 2.2 mmol/L or less (e40 mg/dL),concomitant insulin levels 6 KU/mL or greater (Q36 pmol/L;Q3 KU/mL), C-peptide levels 200 pmol/L or greater, proinsulinlevels 5 pmol/L or greater, A-hydroxybutyrate levels 2.7 mmol/Lor less, and absence of sulfonylurea (metabolites) in the plasma

and/or urine. Further controlled testing under supervision includesthe 72-hour fast, which is the criterion standard for establishingthe diagnosis of insulinoma.37 Actually, 98% of patients withinsulinomas will develop symptomatic hypoglycemia within 72hours.1 When the patient develops symptoms and the blood glu-cose levels are 2.2 mmol/L or less (e40 mg/dL), blood should alsobe drawn for C-peptide, proinsulin, and insulin and the fast shouldbe stopped. Failure in appropriate insulin suppression in thepresence of hypoglycemia substantiates an autonomously secret-ing insulinoma.37 It has been proposed that the sensitivity of the48-hour fasting test is between 94.5% and 95.7% and should beenough for the diagnosis of insulinoma instead of the 72-hour-fast test.38,39 In the case of suspected insulinoma, it is important tokeep in mind the possible differential diagnosis: nesidioblastosis,noninsulinoma pancreatogenous hypoglycemia syndrome (seediscussion later), and multiple adenomas.

Insulinomas have an estimated annual incidence of 1 to4 per million persons. Approximately 4% to 5% of patients withinsulinoma haveMEN I.34 Insulinomas are usually single tumors(except in patients with MEN I), generally small (ie, G1 cm), andalmost always (999%) intrapancreatic in location and, in contrastto all other PNETs, are benign in more than 85% to 90% ofpatients.34,36,37,40

GASTRINOMAPatients typically present with abdominal pain due to peptic

ulcer disease (PUD), diarrhea, and reux esophagitis.41 Zollinger-Ellison syndrome should be suspected in patients with PUDwith diarrhea, with ulcers in unusual locations, with severe PUDor with complications of PUD, with PUD without Helicobacterpylori present, with PUD with a family history of PUD or endo-crinopathies, or with PUD with prominent gastric folds, pres-ence of an endocrinopathy or with hypergastrinemia.29,41Y44 Thediagnosis of ZES requires the demonstration of inappropriatehypergastrinemia (ie, hypergastrinemia present at time of acidhypersecretion).29,41Y44 When ZES is suspected, the initial deter-mination in most centers is a fasting gastrin level, because it willbe elevated in 99% to 100% of ZES patients.29,43,44 The diagnosiscan be complicated by the fact that other conditions, some ofwhich are much more common than ZES, can cause hypergas-trinemia in a range seen with ZES patients.29,41Y46 These otherconditions either can result in hypergastrinemia because of theircausing hypochlorhydria/achlorhydria (atrophic gastritis, PPItreatment) or can be associated with increased acid secretionsimilar to ZES.29,41,43,44,47 Recent studies47,48 show the wide-spread use of PPIs can particularly complicate and delay the di-agnosis of ZES. This occurs because PPIs, in contrast to histamineH2-receptor antagonists, control the symptoms in most ZESpatients with conventional doses used to treat idiopathic PUD/esophagitis and thus mask the diagnosis.29,44,47 Furthermore, PPIscause hypergastrinemia in patients without ZES, commonly in therange that is seen with the majority of ZES patients.29,47Y49 Forthese measures, a washout period from PPIs treatment of 1 weekis recommended.42 If the FSG is 1000 ng/L or greater (pg/mL)and the gastric pH is less than 2.5, the diagnosis is established,1 ifthe patient is normocalcemic and free of pyloric obstruction andhas normal renal function.13 The 2006 European NeuroendocrineTumor Society (ENETS) guidelines had cutoff values of greaterthan 10-fold elevation for FSG and gastric pH 2 or less.42 Toexclude physiological hypergastrinemia (associated with hypo-chlorhydria/achlorhydria), it is essential to measure gastric pHwhen hypergastrinemia is detected because it is 2 or less in allpatients with ZES not taking antisecretory drugs.50 Because ofthe presence of other conditions that can also cause hypergastrin-emia and an acidic pH, the diagnosis of ZES may require the




assessment of basal acid output as well as a secretin provocativegastrin test.29,42,44,45 Gastrinomas ectopically express secretinreceptors, and intravenous administration of secretin character-istically causes an exaggerated release of secretin by the gas-trinoma (Q120-pg/mL increase postsecretin is positive togastrinoma,45 if the patients is not taking PPIs).45,51 In the casethat the FSG values are not high enough to make a denitivediagnosis, then a provocative test should be done. Administrationof secretin after an overnight fast is performed, serum for esti-mation of gastrin levels is collected fasting and 2, 5, 10, 15, and30 minutes after the secretin bolus. In healthy people, the increasein gastrin is not higher than 50% over the baseline level; in thepresence of a gastrinoma, the increase is greater than 100 ng/Labove the baseline levels, which will also distinguish patientswith hypergastrinemia from those with achlorhydric states (ie,type 1 gastric carcinoids, use of PPIs, pernicious anemia, atrophicgastritis), who do not respond to the administration of secretin,unlike patients with a gastrinoma.4

With some patients suspected of having ZES, there can berisk to stopping PPIs to assess fasting gastrin levels and performacid secretory studies or secretin provocative testing; thus, it isbest that these patients be referred to a clinical unit experiencedin making the diagnosis of ZES.

Currently gastrinomas, in contrast to older studies, arefound in the duodenum in more than 60% of patients withsporadic ZES (985% with MEN I/ZES), are usually single insporadic ZES and invariably multiple in MEN I/ZES, are usuallysmall in size in the duodenum (G1 cm), and are malignant in60% to 90% of cases.34,52Y56 Recent studies show pancreatictumors are more aggressive than duodenal tumors, are muchmore likely to metastasize to liver and/or bone, and are morelikely to be present in the 25% of ZES patients with aggressivegastrinomas.34,52Y56

The diagnosis of ZES in patients with MEN I can becomplicated by the fact that the presence or absence of hyper-parathyroidism with the resultant hypercalcemia can have amarked effect of fasting gastrin levels, basal acid output, and thesecretin test results.34,35,60Y62 Each of these parameters canmarkedly decrease after correction of the hyperparathyroidism,by an effective parathyroidectomy (Q3.5 glands removed), andthus can mask the diagnosis of ZES in a MEN I patient.30,31,56Y58

GLUCAGONOMA

Glucagonoma or the Sweet SyndromeDiabetes accompanied by the 4D syndrome (dermatosis:

necrolytic migratory erythema, depression, deep venous throm-bosis, and diarrhea) is the clinical presentation of glucagonomas.Glucagonomas cause glucose intolerance (40%Y90%),weight loss(80%), and a pathognomonic rash characterized by a migratorynecrolytic erythema (70%Y90%).29,33,63Y65

Glucose intolerance in the glucagonoma syndrome may re-late to tumor size. Fasting plasma glucagon levels tend to be higherin patients with large hepatic metastases than in those withouthepaticmetastases,66 and all patients with large hepatic metastaseshave glucose intolerance. Massive hepatic metastases may de-crease the ability of the liver to metabolize splanchnic glucagon,thus increasing peripheral plasma glucagon levels. Glucagon maynot directly induce hyperglycemia, however, unless metabolism ofglucose by the liver is directly compromised. Another factor maybe variation in the molecular species of glucagon that is present ineach case and its biologic potency.10

Glucagonomas are generally single and large tumors (mean,6 cm), associated with liver metastases in more than 60% ofcases at diagnosis, and are almost entirely intrapancreatic in

location.29,33,63Y65 Although glucagonomas may be associatedwith glucose intolerance, clinically signicant hyperglycemiaoccurs in only half of such patients. Patients with glucagon-omas are frequently initially diagnosed by a dermatologist, afterpresenting with necrolytic migratory erythema. This rash,characterized by raised erythematous patches beginning in theperineum and subsequently involving the trunk and extremities,is found in more than two thirds of all patients.29,33,63Y65 How-ever, necrolytic migratory erythema is not specic for gluca-gonomas because it can also occur in cirrhosis, pancreatitis, andceliac disease.29 The diagnosis of a glucagonoma requires thedemonstration of increased plasma glucagon levels (usually500Y1000 pg/mL, normal G50) in the presence of the appropriatesymptoms.29,33,63Y65 In previously reported cases of glucago-noma in which plasma glucagon concentrations were measuredby radioimmunoassay, fasting plasma glucagon concentrationswere 2100 T 334 pg/mL. These levels are markedly higher thanthose reported in normal, fasting subjects (ie, 150 pg/mL) or inthose with other disorders causing hyperglucagonemia, includ-ing diabetes mellitus, burn injury, acute trauma, bacteremia,cirrhosis, renal failure, or Cushing syndrome, where fastingplasma glucagon concentrations often are elevated but less than500 pg/mL.

As with other islet cell neoplasms, glucagonomas mayoverproduce multiple hormones such as insulin, ACTH, PP,parathyroid hormone (PTH) or substances with PTH-like ac-tivity, gastrin, 5-HT, VIP, and melanocyte-stimulating hormone,in that order of frequency.67

VIPOMASVIPomas are also called Verner-Morrison syndrome, pan-

creatic cholera, WDHA syndrome (for watery diarrhea, hypo-kalemia, and achlorhydria). VIPomas are PNETs ectopicallysecreting vasoactive intestinal polypeptide (VIP), which leads toprofound, large-volume diarrhea (100%9700 mL/day), hypo-kalemia, and achlorhydria.29,31,33,63,68 VIPomas are usuallysingle tumors, metastatic at presentation in 70% to 80% of casesand in adults are intrapancreatic in location in more than 95% ofcases, whereas in children they often are ganglioneuromas/ganglioblastomas.29,31,33,63,68 The diagnosis requires the dem-onstration of an elevated plasma VIP level in a patient with large-volume secretory diarrhea.29,31,33,63,68

SOMATOSTATINOMASomatostatin (SRIF) is a tetradecapeptide that inhibits nu-

merous endocrine and exocrine secretory functions. Almost allgut hormones that have been studied are inhibited by SRIF, in-cluding insulin, PP, glucagon, gastrin, secretin, GIP, and moti-lin.69 In addition to inhibition of the endocrine secretions, SRIFhas direct effects on a number of target organs.70 For example, itis a potent inhibitor of basal and PG-stimulated gastric acidsecretion. It also has marked effects on GI transit time, intestinalmotility, and absorption of nutrients from the small intestine. Themajor effect in the small intestine seems to be a delay in theabsorption of fat and reduced absorption of calcium.

The salient features of the somatostatinoma syndrome arediabetes, diarrhea or steatorrhea, gallbladder disease, hypo-chlorhydria, and weight loss.71Y73 The rst cases of the soma-tostatinoma syndrome were reported in 1977 by Ganda andcolleagues.71 We have examined the cases reported since 1977and describe here the features now recognized to be a part ofthe syndrome. For convenience, we have divided the casesinto those arising from the pancreas, the intestine, and extra-pancreatic tumors. It seems that the syndrome differs among




tumors arising from the pancreas and the intestine or extra-pancreatic sites. Therefore, these will be considered separately.

Most patients were between 40 and 60 years of age. There isa 2:1 ratio of female to male patients, which contrasts with theequal-sex incidence for other islet cell tumors.74

PLASMA SOMATOSTATIN-LIKEIMMUNOREACTIVITY

The mean somatostatin-like immunoreactivity (SLI) con-centration in patients with pancreatic somatostatinoma was50 times higher than normal (range, 1Y250 times). Intestinalsomatostatinomas, however, had only slightly elevated or normalSLI concentrations.

DIABETES MELLITUS AND HYPOGLYCEMIASeventy-ve percent of patients with pancreatic tumors had

diabetes mellitus. In contrast, diabetes occurred only in 11% ofpatients with intestinal tumors. In all instances, the diabetes wasrelatively mild and could be controlled with diet and/or oralhypoglycemic agents or with small doses of insulin. It is notclear, however, whether the differential inhibition of insulin anddiabetogenic hormones can explain the usually mild degree ofdiabetes and the rarity of ketoacidosis in patients with soma-tostatinoma. Replacement of functional islet cell tissue by pan-creatic tumor may be another reason for the development ofdiabetes in most patients with pancreatic somatostatinoma,contrasting with the low incidence in patients with intestinaltumors. These tumors usually are large and therefore destroysubstantial portions of the pancreas.

GALLBLADDER DISEASEFifty-nine percent of patients with pancreatic tumors and

27%of patientswith intestinal tumors had gallbladder disease. Thehigh incidence of gallbladder disease in patients with somatosta-tinoma and the absence of such an association in any other islet celltumor suggest a causal relationship between gallbladder diseaseand somatostatinoma. Infusion of somatostatin into normal humansubjects has been shown to inhibit gallbladder emptying,70,75

suggesting that somatostatin-mediated inhibition of gallbladderemptying may cause the observed high rate of gallbladder diseasein patients with somatostatinoma. This thesis is supported bythe observation of massively dilated gallbladders without stonesor other pathology76,77 in patients with somatostatin-secretingtumors.

DIARRHEA AND STEATORRHEADiarrhea consisting of 3 to 10 frequently foul-smelling stools

per day and/or steatorrhea from 20 to 76 g of fat per 24 hours iscommon in patients with pancreatic somatostatinoma. This couldresult from the effects of high levels of somatostatin within thepancreas, serving as a paracrine mediator to inhibit exocrine se-cretion or, alternatively, from the somatostatinomas causingduct obstruction. In some cases, the severity of diarrhea and ste-atorrhea parallels the course of the disease, worsening as the tumoradvances and metastatic disease spreads and improving aftertumor resection. Somatostatin has been shown to inhibit thepancreatic secretion of proteolytic enzymes, water, bicarbonate,78

and gallbladder motility.79 In addition, it inhibits the absorption oflipids.80 All but 1 patient with diarrhea and steatorrhea had highplasma somatostatin concentrations. The rarity of diarrhea and/orsteatorrhea in patients with intestinal somatostatinomasmay resultfrom the lower SLI levels.

HYPOCHLORHYDRIAInfusion of somatostatin has been shown to inhibit gastric

acid secretion in human subjects.81 Thus, hypochlorhydria inpatients with somatostatinoma in the absence of gastric mucosalabnormalities likely results from elevated somatostatin con-centrations. Basal and stimulated acid secretion was inhibited in87% of patients with pancreatic tumors tested but in only 12% ofpatients with intestinal tumors.

WEIGHT LOSSWeight loss ranging from 9 to 21 kg over several months

occurred in one third of patients with pancreatic tumors and onefth of patients with intestinal tumors. The weight loss mayrelate to malabsorption and diarrhea, but in small intestinaltumors, anorexia, abdominal pain, and yet unexplained reasonsmay be relevant.

ASSOCIATED ENDOCRINE DISORDERSOf great interest is the presence of cafe au lait spots, neu-

robromatosis, and paroxysmal hypertension in patients withintestinal tumors. Thus, approximately 50% of all patients haveother endocrinopathies in addition to their somatostatinoma.Occurrence of MEN I has been recognized in patients with isletcell tumors, and MEN II-A or MEN II-B syndromes are presentin association with pheochromocytomas and neurobromatosis,respectively. It seems that an additional dimension of the ductassociated tumors is MEN II. Secretion of different hormones bythe same islet cell tumor, sometimes resulting in 2 distinctclinical disorders, is now being recognized with increasing fre-quency.82 These possibilities should be considered during en-docrine workups of patients with islet cell tumors and theirrelatives.

TUMOR LOCATIONOf the reported primary tumors, 60% were found in the

pancreas and 40% in the duodenum or jejunum. Of the pancre-atic tumors, 50% were located in the head and 25% in the tail,and the remaining tumors either inltrated the whole pancreas orwere found in the body. Regarding extrapancreatic locations,approximately 50% originate in the duodenum, approximately50% originate in the ampulla, and, rarely, one is found in thejejunum. Thus, approximately 60% of somatostatinomas origi-nate in the upper intestinal tract, which probably is a conse-quence of the relatively large number of normal D cells in thisregion.

TUMOR SIZESomatostatinomas tend to be large, similar to glucagono-

mas,83 but unlike insulinomas and gastrinomas, which, as a rule,are small.84Y86 Within the intestine, tumors have tended to besmaller. Symptoms associated with somatostatinomas and glu-cagonomas are less pronounced and probably do not developuntil very high blood levels of the respective hormones havebeen attained. As a result, somatostatinomas and glucagonomasare likely to be diagnosed later.

INCIDENCE OF MALIGNANCYEighty percent of pancreatic patients with pancreatic

somatostatinomas were metastatic at diagnosis, and 50% withintestinal tumors had evidence of metastatic disease. Metastasisto the liver is most frequent, and regional lymph node involve-ment and metastases to bone are less so. Thus, in approximately




70% of cases, metastatic disease is present at diagnosis. This issimilar to the high incidence of malignancy in glucagonoma84

and in gastrinoma,85 but it is distinctly different from the lowincidence of malignant in insulinoma.86 The high prevalence ofmetastatic disease in somatostatinoma also may be a conse-quence of late diagnosis but apparently is not dependent on thetissue of origin.

MICROSCOPIC APPEARANCEOn light microscopy, most tumors appear to be well-

differentiated islet cell or carcinoid-type tumors. Some show amixed picture, consisting of separate zones of differentiated andanaplastic cells. In the differentiated areas, cells are arranged inlobular or acinar patterns that are separated by brovascularstroma. Less well-differentiated areas consist of sheets of cellsinterrupted by brous septa.

Diffuse positive immunoreactivity for somatostatin usuallyis found, which contrasts with the rarity of somatostatin-positivecells in gastrinomas and other tumors. There is a unique oc-currence of psammoma bodies in somatostatinomas localizedwithin the duodenum. In addition, there is abundant immuno-logic evidence for the presence of cells containing insulin, cal-citonin, gastrin and VIP, ACTH, prostaglandin E2, and SP. Intumors with multiple hormones, however, SLI-containing cellsrepresent the large majority of all cells containing hormonesdetected by immunopathology.

SOMATOSTATIN-CONTAINING TUMORSOUTSIDE THE GI TRACT

Somatostatin has been found in many tissues outside the GItract. Prominent among those are the hypothalamic and extra-hypothalamic regions of the brain, the peripheral nervous system(including the sympathetic adrenergic ganglia), and the C cellsof the thyroid gland. Not surprisingly, therefore, high con-centrations of somatostatin have been found in tumors origi-nating from these tissues. Sano and colleagues87 and Saito andcolleagues88 reported 7 patients with MTC who had high basalplasma SLI concentrations and high tumor SLI concentrations.Roos and colleagues89 reported elevated plasma SLI con-centrations in 3 of 7 patients with MTC and high tissue SLIconcentrations in 3 of 5 MTC tumors. Some, but not all, of thesepatients exhibited the clinical somatostatinoma syndrome.

Elevated plasma SLI concentrations also have been re-ported in patients with small cell lung cancer.89 One case ofmetastatic bronchial oat cell carcinoma caused Cushing syn-drome, diabetes, diarrhea, steatorrhea, anemia, and weight lossand had a plasma SLI concentration 20 times greater than nor-mal.90 A patient with a bronchogenic carcinoma presenting withdiabetic ketoacidosis and high levels of SLI (95000 pg/mL) hasbeen reported.91 Pheochromocytomas82,92 and catecholamine-producing extra-adrenal paragangliomas86 are other examples ofendocrine tumors producing and secreting somatostatin in ad-dition to other hormonally active substances. One quarter of 37patients with pheochromocytomas had elevated SLI levels.89

DIAGNOSISIn the reported series cited, somatostatinomas often were

found more or less accidentally. In most cases, the tumors werefound either during exploratory laparotomy or upper GI radio-graphic studies, computed tomography (CT), or ultrasound, orendoscopy performed because of various symptoms, includingunexplained abdominal pain, melena, hematemesis, persistentdiarrhea, or in search of insulinomas or ACTH-secreting tumors.

Once found, the tumors were identied as somatostatinoma bythe demonstration of elevated tissue concentrations of SLI and/orprevalence of D cells by immunocytochemistry or demonstrationof elevated plasma SLI concentrations. Thus, events leading tothe diagnosis of somatostatinoma usually occur in reverse order.In other islet cell tumors, the clinical symptoms and signs usuallysuggest the diagnosis, which then is established by demonstra-tion of diagnostically elevated blood hormone levels, followingwhich efforts are undertaken to localize the tumors. It can beexpected that the same sequence of diagnostic procedures will befollowed in the future for the diagnosis of somatostatinoma,mainly for 2 reasons: (a) the increasing familiarity of physicianswith the clinical somatostatinoma syndrome (this symptomcomplex, although not pathognomonic, is nevertheless suf-ciently characteristic of somatostatinoma to suggest the correctdiagnosis) and (b) the greater availability of reliable radio-immunoassays for the determination of SLI in blood has in-creased the yield. Presently, these assays are complicated by theneed for cumbersome extraction procedures and are not readilyavailable. (Assay available at Inter Science Institute: 800-255-2873.) It should be recognized, however, that the syndrome israre. Of 1199 cases screened for somatostatinoma at the Uni-versity of Michigan between 1982 and 1986, only 8 cases haddiagnostic serum levels.

The diagnosis of somatostatinoma at a time when bloodSLI concentrations are normal or only marginally elevatedrequires reliable provocative tests. Increased plasma SLI con-centrations have been reported after intravenous infusion oftolbutamide and arginine, and decreased SLI concentrationshave been observed after intravenous infusion of diazoxide.Arginine is a well-established stimulant for normal D cells andthus is unlikely to differentiate between normal and supra-normal somatostatin secretion. The same may be true fordiazoxide, which has been shown to decrease SLI secretionfrom normal dog pancreas as well as in patients with soma-tostatinoma.90 Tolbutamide stimulates SLI release from normaldog and rat pancreas,78,79,93 but no change was found in thecirculating SLI concentrations of 3 normal human subjectsafter intravenous injection of 1 g of tolbutamide.94 Therefore,at present, tolbutamide seems to be a candidate for a provoc-ative agent in the diagnosis of somatostatinoma, but its reli-ability must be established in a greater number of patients andcontrols. Until then, it may be necessary to measure plasma SLIconcentrations during routine work-ups for postprandial dys-pepsia and gallbladder disorders,72 for diabetes in patientswithout a family history, and for unexplained steatorrhea asthese ndings can be early signs of somatostatinomas.

Somatostatinomas are PNETs that can occur in the duo-denum or pancreas.29,33,63,95,96

OTHER RARE FUNCTIONAL PNETSGRFomas ectopically secrete GH-releasing factor, which

results in acromegaly, which is generally clinically indistinguish-able from that caused by pituitary adenomas.29,63,97 GRFomas inthe pancreas are generally single, large tumors at diagnosis; onethird have liver metastases, and they are found in the pancreasin 30% of cases, 54% in the lung, and the remainder primarily inother abdominal locations.29,63,97 Pancreatic NETs causing hy-percalcemia usually secrete PTH-related peptide (PTH-rP) as wellas other biologically active peptides and are similar to pancreaticACTHomas associated with ectopic Cushing syndrome in thatboth are usually large tumors at diagnosis, with 80% to 90% asso-ciated with liver metastases.29,63,98,99 With ACTHomas or PNETscausing hypercalcemia, the diagnosis is made by the presence ofa PETwith the appropriated elevated hormonal assay result.




NONFUNCTIONING PANCREATICENDOCRINE TUMORS

Nonfunctional PNETs are intrapancreatic in location, char-acteristically large (70% are 95 cm), and at an advanced stagewhen rst diagnosed, with 60% to 85% having liver metastasesin most series.29,31,32,63,100 Because NF-PNETs are not associ-ated with a clinical hormonal syndrome, they present clinicallywith symptoms due to the tumor per se, which include primarilyabdominal pain (40%Y60%), weight loss, or jaundice.29,31,32,63,100

In recent years, NF-PNETs are increasingly being discoveredby chance on imaging studies being performed for various non-specic abdominal symptoms.29,101 Even though NF-PNETs donot secrete peptides causing a clinical syndrome, they are notbiologically inactive, because they characteristically secrete anumber of other peptides, which are frequently used in their diag-nosis. These include chromogranins, especially CgA (70%Y100%)and PP (50%Y100%).29,31,32,63,100 The presence of an NF-PET issuggested by the presence of a pancreatic mass in a patient with-out hormonal symptoms, who has an elevated serum PP or CgAlevel or a positive OctreoScan (Mallinckrodt Medical, Inc., St.Louis, MO) (somatostatin receptor scintigraphy [SRS]) (discussedin the next section). It is important to remember that an elevatedPP level or CgA level is not specic for NF-PNETs.29,31,32,63,100

MISCELLANEOUS PANCREATIC NETSForVIPomas, glucagonomas, somatostatinomas, and PPomas,

the biochemical markers are VIP, glucagon, somatostatin, andPP, respectively.1 For every PNET, always screen for MEN type Isyndrome measuring ionized calcium, serum PTH, and prolac-tin.102 Biochemical screening for PNETs, in the presence ofsuspected MEN I syndrome, can include gastrin, insulin/proin-sulin, PP, glucagon, and CgA, together having a sensitivity ofapproximately 70%, which can be increased if >-hCG and A-hCGsubunits, VIP, and postprandial gastrin and PP measurements areadded.13

Acromegaly or GigantismAcromegaly or gigantism can present when any NET

secretes GH or GH-releasing hormone (GHRH). Basal levels ofGH and IGF-1 are usually enough to make a diagnosis; but in15% to 20% of the patients, further investigation is needed toshow nonsuppressibility of GH to an oral glucose tolerance test,a somatostatin inhibition test, or a bromocriptine suppressiontest. In the case of the oral glucose tolerance test, also measurelipids and insulin, which should also be suppressed. Other pi-tuitary and hypothalamic hormones should also be measured,such as prolactin, the > and A subunits of gonadotropins, andthyroid-stimulating hormone.4

Cushing SyndromeA pituitary tumor, small cell carcinoma of the lung (known

to produce ACTH), or an ACTH-secreting NET will presentclinically as Cushing syndrome from oversecretion of cortisol,adrenal androgens, and 11-deoxycorticosterone. To reach thediagnosis, several steps should be followed. New guidelinesfor the diagnosis of Cushing syndrome have been published,although some of the recommendations are based on low-qualityevidence. Their proposed approach is as follows.

After excluding exogenous glucocorticoid use (iatrogenicCushing syndrome), patients with unusual features for age such asosteoporosis or hypertension, patients with multiple and progres-sive features predictive of Cushing syndrome (easy bruising, facialplethora, proximal myopathy or muscle weakness, reddish/purplestriae, weight gain in children with decreasing growth velocity),

and patients with adrenal incidentaloma compatiblewith adenomashould undergo testing for Cushing syndrome starting with 1 testwith high diagnostic accuracy: urine free cortisol (at least 2 mea-surements), late night salivary cortisol (2 measurements), 1 mgovernight dexamethasone suppression test, or longer low-dosedexamethasone suppression test (2 mg/d for 48 hours). If the testis negative, and the pretest probability was low, then follow-up in6 months is recommended for progression of symptoms; in caseof a negative test but with a high pretest probability, more than1 test should be performed. In some cases, a serum midnightcortisol or dexamethasone-corticotropin releasing hormone testshould be done.103

BIOCHEMICAL ASSESSMENT ANDMONITORING FOR PNETS

Specic hormonal assays are needed to establish the diag-nosis of each functional PET as outlined in the discussion ofeach tumor type in Pancreatic NETs. Specically, for insulino-mas, an assessment of plasma insulin, proinsulin, and C-peptideis needed at the time of glucose determinations, usually during afast.29,37,104 For ZES, serum gastrin is needed either alone orduring a secretin provocation test.29,37,43,45,104 For VIPomas, aplasma VIP level is needed; for glucagonoma, plasma glucagonlevels; for GRFomas, plasma GH and GRF levels; for Cushingsyndrome, urinary cortical, plasma ACTH, and appropriateACTH suppression studies; for hypercalcemia with PET, bothserum PTH levels and PTH-rP levels are indicated; and for aPET with carcinoid syndrome, urinary 5-H1AA should bemeasured.29,33,63,104,105 Plasma CgA can be used as a marker inpatients with both functional and nonfunctional pancreatic en-docrine tumors.29,104Y106 Chromogranin A should be used withcaution as a marker of disease activity in patients treated withsomatostatin analogs, because these agents signicantly reduceplasma CgA levels, a change that may be more reective ofchanges in hormonal synthesis and release from tumor cells thanan actual reduction in tumor mass.17,105 In patients on stabledoses of somatostatin analogs, consistent increases in plasmaCgA levels over time may reect loss of secretory control and/ortumor growth.18,40,104Y106

PHEOCHROMOCYTOMASThe main signs and symptoms of catecholamine excess in-

clude hypertension, palpitations, headache, sweating, and pallor.Less common signs and symptoms are fatigue, nausea, weightloss, constipation, ushing, and fever. According to the degreeof catecholamine excess, patients may present with myocardialinfarction, arrhythmia, stroke, or other vascular manifestations(eg, any organ ischemia). Similar signs and symptoms are pro-duced by numerous other clinical conditions, and therefore,pheochromocytoma is often referred to as the Bgreat mimic.[

In general, about 80% of pheochromocytomas are locatedin the adrenal medulla.107 Extra-adrenal sympathetic para-gangliomas in the abdomen most commonly arise from chro-mafn tissue around the inferior mesenteric artery (the organ ofZuckerkandl) and aortic bifurcation, less commonly from anyother chromafn tissue in the abdomen, pelvis, and thorax.108

Extra-adrenal parasympathetic paragangliomas are most com-monly found in the neck and head.

Pheochromocytomas and sympathetic extra-adrenal para-gangliomas almost all produce, store, metabolize, and secretecatecholamines or their metabolites. Head and neck paragan-gliomas, however, rarely produce signicant amounts of cate-cholamines (G5%).




EPIDEMIOLOGYPheochromocytomas and paragangliomas are rare and occur

in about 0.05% to 0.1% of patients with sustained hypertension.However, this probably accounts for only 50% of people harbor-ing pheochromocytoma or paraganglioma because about halfthe patients with pheochromocytoma or paraganglioma haveparoxysmal hypertension or normotension. The prevalence ofpheochromocytoma and paraganglioma can be estimated to liebetween 1:6500 and 1:2500, with the annual incidence in theUnited States of 500 to 1600 cases per year.

PATHOLOGY AND MOLECULAR GENETICSMalignancy is dened by the presence of metastases. At

present, it is estimated that at least 24% to 27% of pheochro-mocytomas or paragangliomas are associated with known ge-netic mutations; in children, this prevalence may be as high as40%.109Y114

Pheochromocytomas may occur sporadically or as part ofhereditary syndrome. According to the latest studies, amongpatients with nonsyndromic pheochromocytoma, up to about24% of tumors may be hereditary.109,114,115 Hereditary pheo-chromocytoma is associated with MEN IIA or MEN IIB, NF-1,von Hippel-Linda (VHL) syndrome, and familial paragangliomasand pheochromocytomas due to germ-line mutations of genesencoding succinate dehydrogenase subunits B, C, and D (SDHB,SDHC, and SDHD). In general, the traits are inherited in an au-tosomal dominant pattern.115

Specically, MEN I- and NF-1Yrelated pheochromocytomaalways secrete epinephrine, VHL-related pheochromocytomasalways secrete norepinephrine, and elevation of dopamine to-gether with norepinephrine is seen in some SDHB-relatedparagangliomas. In contrast to MEN II, VHL, and NF-1 tumorsthat are almost always found in the adrenal gland, SDHB-relatedtumors are found in extra-adrenal localizations. In those patientswith malignant disease secondary to an extra-adrenal para-ganglioma, almost 50% had SDHB mutations.116 Some studiessuggested that more than two thirds of patients with SDHB-related pheochromocytoma or paraganglioma will developmetastatic disease.117,118 Family history is often helpful inMEN II, VHL, and NF-1 tumors, but only 10% of the currentlyinvestigated patients with SDHB mutations have a positivefamily history for pheochromocytoma or paraganglioma.117

BIOCHEMICAL MARKERS FORPHEOCHROMOCYTOMAS AND

PARAGANGLIOMASIn line with these concepts, numerous independent studies

have now conrmed that measurements of fractionated meta-nephrines (ie, normetanephrine and metanephrine measuredseparately) in urine or plasma provide superior diagnosticsensitivity over measurement of the parent catecholamines(Table 3).119Y121 However, to preserve high diagnostic sensi-tivity, it is strongly recommended to obtain blood samples inthe supine position.122

Current recommendations are that initial testing for pheo-chromocytoma or paraganglioma must include measurementsof fractionated metanephrines in plasma, urine, or both, as avail-able.123 Blood sampling should be performed at a supine positionafter about 15 to 20 minutes of intravenous catheter insertion.Food, caffeinated beverages, strenuous physical activity, or smok-ing are not permitted at least about 8 to 12 hours before the test-ing. The elevation of plasma metanephrines of more than 4-foldabove the upper reference limit is associated with close to 100%probability of the tumor.124

Should additional biochemical testing be necessary, thepossibility of false-positive results due to medications, clini-cal conditions (as described above), or inadequate samplingconditions (eg, blood sampling while seated) should rstbe considered and eliminated.124 In patients with plasmametanephrine values above the upper reference limit and lessthan 4-fold above that limit, the clonidine suppression testcombined with measurements of plasma catecholamines andnormetanephrine may prove useful.124 Guller et al125 publishedin 2006 that the tests of choice to establish the diagnosis ofpheochromocytomas are urinary normetanephrine and plateletnorepinephrine with sensitivities of 96.9 and 93.8%, respec-tively. In a study conducted in Switzerland by Giovanella et al126

in 2006, plasma metanephrines and CgA showed 95% sensi-tivity with comparable high specicity and diagnostic accuracy(96% and 96% for CgA, 94% and 95% for MN, respectively). Ifboth were used, then sensitivity increased to 100%. The differencefound between these 2 markers is that only CgA was correlatedwith tumor mass. In 2008, Bilek et al127 also studied the use ofCgA for pheochromocytoma and found that it is a great markerfor following response to treatment and that the levels of CgAwere correlated with the size and the malignancy of the tumor.

Paragangliomas are NETs that arise from the paravertebralaxis. Sympathetic paragangliomas usually hypersecrete cate-cholamines and are localized in the thorax, abdomen, or pelvis.Parasympathetic paragangliomas are nonsecretory tumors usu-ally localized in the head and neck area.128

Diagnosis of paragangliomas is similar to that of pheochro-mocytomas because these 2 entities differ only in their placeof origin, extra-adrenal versus adrenal, respectively. Algeciras-Schimnich et al129 suggested that when plasma fractionatedmetanephrines are measured and values are not 4-fold aboveupper normal limit, then serum or plasma CgA and urine frac-tionated metanephrines should be measured to conrm the diag-nosis. After surgery, the biochemical follow-up should be done1 to 2 weeks later with 24-hour urine fractionated catecholaminesand metanephrines; if normal, complete resection is claimed, butif it persists elevated, a second primary or occult metastasisshould be suspected and investigated. Young130 also proposed anannual biochemical testing follow-up for life, with 24-hour uri-nary excretion of fractionated catecholamines and metanephrinesor plasma fractionated metanephrines, and only in the case ofelevated levels that imaging follow-up is considered.

All patients with paragangliomas should be consideredfor genetic testing with VHL, RET, NF1, SDHD, SDHB, andSDHC genes.128 If positive, then rst-degree relatives genetictesting should be suggested, and genetic counseling should beoffered. First-degree relatives should always undergo biochem-ical testing with 24-hour urine fractionated metanephrines andcatecholamines.130

Medullary Carcinoma of the ThyroidThese originate from the parafollicular cells of the thyroid,

which secrete calcitonin. They represent 4% to 10% of all thyroidneoplasms.131 Medullary carcinoma of the thyroid can present as2 different forms, sporadic (75%) or inherited (25%), and the lastcan be either isolated or part of the MEN II syndrome.132 A germ-line autosomal dominant mutation in the RET protooncogene,which encodes for a transmembrane tyrosine kinase receptor,predisposes individuals to develop MCT. Screening for RETgerm-line mutations has allowed for early and accurate diagnosisof patients at risk of developing MCT.133,134

The most common clinical presentation of MCT is a thy-roid nodule, either single or as multinodular goiter. Usually, noother manifestations are present unless the tumor is already in




stage IV (metastatic disease), when diarrhea and/or ushingcan present.135

The calcitonin secreting nature of these tumors and the factthat calcitonin is almost exclusively secreted by C cells explainwhy this hormone is the preferred biochemical marker for thediagnosis and follow-up of this disease; besides, it has beenshown that calcitonin measurement is more sensitive than ne-needle aspiration for the diagnosis of MCT.135 A 10-year sur-vival of only 50% for MTC patients is reported in several series.The only possible means to improve the cure and survival rateof these patients consists in early diagnosis and early surgicaltreatment, while the MTC is still intrathyroid.135 Costanteet al131 reported on 2007 that the positive predictive value ofbasal calcitonin levels greater than 100 pg/mL is 100% for MCT,and if PG stimulation test is used, calcitonin levels greater than100 pg/mL had a positive predictive value of 40%, but below thiscutoff value, the false-positive results increase until the positivepredictive value of basal calcitonin levels greater than 20 pg/mLis less than 25%. Cohen et al132 found that calcitonin levels arenot only useful as diagnostic marker, but are also correlated withtumor size and metastasis, which gives some prognostic value tothis hormone. When levels are less than 50 pg/mL preopera-tively, the normalization of calcitonin levels postoperatively isfound in 97.8% of the patients. Scheuba et al136 recently pub-lished that values of basal calcitonin greater than 64 pg/mL orstimulated calcitonin levels greater than 560 pg/mL had a sen-sitivity of 100% for MCT. Calcitonin increase can be observedalso in parafollicular C-cell hyperplasia (CCH) and otherextrathyroidal conditions. The PG test is used to distinguishMCT from CCH because it is thought that the response to thisstimulus is typical of pathological thyroid C cells. The cutoffvalue of calcitonin response between patients with MTC andCCH remains to be established.137 Pentagastrin stimulation testis no longer available in the United States, but it consisted of theintravenous injection of 0.5 Kg PG/kg body weight and mea-surements of calcitonin at 0, 1, 2, 5, and 10 minutes after theinjection; healthy people do not experience an increase in cal-citonin greater than 200 pg/mL after the administration of PG.4

Instead, calcitonin provocation can also be accomplished with anintravenous calcium infusion; however, there is a signicantdanger of cardiac arrhythmias.

C-Cell HyperplasiaThis entity has been proposed to be a precancerous lesion

that eventually transforms into MCT. Schley et al138 submitted astudy where 3 cases are reported in which patients presentedwith ushing, abdominal pain, diarrhea, and facial telangiecta-sia, resembling carcinoid syndrome, but the only biochemicalabnormalities were elevated calcitonin levels and positivePG and calcium infusion tests. Venous sampling was performed,and it localized the overproduction of calcitonin to the thyroid,and histology showed CCH. After thyroidectomy, symptomsresolved and calcitonin levels returned to normal. They proposedthat the condition might be a gene mutation, but so far, the sitehas not been identied considering that RET protooncogene wasnegative in the 3 patients. These ndings suggest that every caseof ushing and diarrhea should have a calcitonin measurement,considering CCH or MCT in the differential diagnosis.

Multiple Endocrine Neoplasia SyndromesThis entity is classied as MEN either type I or II. They

are both inherited in an autosomal dominant pattern. Mutationson the MEN I tumor suppressor gene (inactivated) or the RETprotooncogene (activated) are found in MEN types I and II,respectively.139

MEN Type IMultiple endocrine neoplasia type I is characterized by

hyperplasia and/or neoplasm of the parathyroid glands, entero-PNETs, and pituitary adenomas. Some patients do not presentwith all these tumors, so it has been agreed that diagnosis ismade when a patient presents with 2 of these concomitantly. Todiagnose familial MEN I syndrome, a rst-degree relative hasto present at least 1 of the tumors mentioned above.140 Hyper-parathyroidism occurs in about 90% of patients; endocrinepancreatic tumors in 60% of patients. Usually they are small andnonfunctional; the most common hormonally active ones areinsulinomas or gastrinomas. Pituitary adenomas are present in40% of patients, and in 60% of the patients, skin manifestationscan also be present.140,141 Genetic studies are available for MENtype I syndrome; MEN I germ-line mutations are found in thesepatients, but its presence does not prompt any immediate in-tervention.142 Piecha et al140 proposed a recommendation forcarriers of MEN I mutation to be screened biochemically every1 to 3 years for hyperparathyroidism, prolactinoma, gastrinoma,insulinoma, and other enteropancreatic tumors.

MEN Type IIThis syndrome is subclassied into type IIA, IIB, and fa-

milial MCT, all sharing the presence of MCT; and they are allcharacterized by an activating germ-line mutation in the RETprotooncogene, specic for each type and which can be identi-ed in almost 100% of the patients, with genetic testing. Oncethe genetic test demonstrates the mutation, a total thyroidectomyis mandatory either prophylactically in carriers or as treatment inpatients who already present with manifestations of the syn-drome.143 Multiple endocrine neoplasia type IIA presents withMCT, bilateral pheochromocytomas, and primary hyperpara-thyroidism; lately, it has been published that Hirschsprung dis-ease could also be a manifestation of this syndrome, and geneticscreening for RET protooncogene mutation is recommended inthis patients144; MEN type IIB is an association of MCT, pheo-chromocytomas, and mucosal neuromas145; these patients usuallypresent with a marfanoid phenotype.

The biochemical studies recommended for these syndromesare the same as previously proposed for each tumor type, depend-ing on the clinical syndrome, and in the case when MEN syn-drome is suspected, genetic testing should also be performed in thepatient and if positive rst-degree relatives should also be tested.

CLASSIFICATION OF THE BIOCHEMICALMARKERS ACCORDING TO THEIR USE (TABLE 4)

Diagnostic

Chromogranins A and BBoth are part of the granin family. They are stored and

secreted from vesicles present in the neuroendocrine cells, to-gether with other peptides, amines, and neurotransmitters.146

Chromogranin A is the best studied147 and most used, but CgA isnot perfect. Stridsberg et al148 reported there are some commonconditions that can increase the levels of this marker and givefalse-positive measurements including decreased renal functionand treatment with PPIs and even essential hypertension149;these problems are not seen with CgB, so they proposed mea-surement of CgB as a complement to CgA.148

The most important characteristic of these markers is thatthey are secreted not only by the functional tumors, but also bythose less well-differentiated NETs that do not secrete knownhormones.2




High CgA has been shown to be increased in 50% to 100%of patients with NETs.150 Chromogranin A levels may be associ-ated with the primary tumor (gastrinomas 100%, pheochromocy-tomas 89%, carcinoid tumors 80%, nonfunctioning tumors of theendocrine pancreas 69%, and medullary thyroid carcinomas 50%).In addition, blood levels depend upon tumor mass, tumor burden,or progression and malignant nature of the tumor.127,151 Smalltumors may be associated with normal CgA levels.

Sensitivity and specicity of CgA depend on many factors.For example, sensitivity varies from 77.8% to 84% and speci-city from 71.3% to 85.3%, depending on the assay used, andof great importance is to establish the cutoff value that givesthe highest sensitivity without compromising the specicity.152

Another utility of CgA is to discriminate between patients withor without metastasis, which also depends on the assay and thecutoff values used, with a sensitivity of 57% to 63.3% andspecicity 55.6% to 71.4%.152

Pancreatic PolypeptidePancreatic polypeptide is considered another nonspecic

biochemical marker. In a study conducted by Panzuto et al153 inRome, Italy, in 2004, PP sensitivity was 54% in functioningtumors, 57% in nonfunctioning, 63% in pancreatic tumors, and53% in GI tumors. Specicity was 81% compared with disease-free patients, and 67% compared with nonendocrine tumorspatients. But when combined with CgA, the sensitivity increasedcompared with either of the markers alone. When used in com-bination, the sensitivity of these markers is 96% for gastroentero-PNETs; for nonfunctioning tumors, 95%; and for pancreatictumors, 94%.153

Neuron-Specic EnolaseNeuron-specic enolases are enzymes that occur mainly in

cells of neuronal and neuroectodermal origin. Neuron-specic

enolase has been found in thyroid and prostatic carcinomas, neu-roblastomas, small cell lung carcinoma, carcinoids, gastroentero-PNETs, and pheochromocytomas. Despite its high sensitivity(100%), its use is limited as a blood biochemical marker for NETsbecause of its very low specicity (32.9%).26

DETERMINING PROGNOSIS

Chromogranin AOther than the applications of CgA previously discussed,

this marker can be used for prognosis and follow-up. Jensenet al154 found that a reduction on CgA levels of 80% or greaterafter cytoreductive surgery for carcinoid tumors predicts symp-tom relief and disease control; it is associated with improvedpatient outcomes, even after incomplete cytoreduction.

PancreastatinOne of the posttranslational processing products of CgA has

found to be an indicator of poor outcome when its concentrationin plasma is elevated before treatment in patients with NETs. Alevel of greater than 500 pmol/L is an independent indicator ofpoor outcome. This marker is also known to correlate with thenumber of liver metastasis, so it would be appropriate to use it inthe follow-up ofNET patients. Furthermore, Stronge et al155 foundthat an increase in pancreastatin levels after somatostatin-analogtherapy is associated with a poor survival. Other studies haveshown that pancreastatin should be measured before treatment andmonitored during and after it. Plasma levels of this marker greaterthan 5000 pg/mL before treatment were associated with increasedperiprocedure mortality in patients with NETs that underwenthepatic artery chemoembolization.156

These observations suggest that pancreastatin is potentiallya very useful marker not only for diagnosis but more importantlyalso for monitoring treatment response.

TABLE 4. Specic Biochemical Markers for Each Tumor Type211

Site Tumor Type Marker Specificity

All CgA and CgB HighPP, NSE, neurokinin, Neurotensin IntermediateHCG > and A Low

Thymus Foregut Carcinoid ACTH IntermediateBronchus Foregut carcinoid, small cell lung ACTH, ADH, Serotonin, 5-HIAA, Intermediate

carcinoma Histamine, GRP, GHRH, VIP, PTH-rP LowStomach Foregut carcinoid, gastrinoma, Histamine, gastrin Intermediate

ghrelinoma Ghrelin LowPancreas Gastrinoma, insulinoma, glucagonoma,

somatostatinoma, PPoma, VIPomaGastrin, insulin, proinsulin, glucagon,somatostatin

High

C-peptide, neurotensin, VIP, PTH-rP,calcitonin

Low

Duodenum Gastrinoma, somatostatinoma. Somatostatin, gastrin HighIleum Midgut carcinoid Serotonin, 5-HIAA High

Neurokinin A, neuropeptide K, substance P IntermediateColon and rectum Hindgut carcinoid Peptide YY, somatostatin IntermediateBone Metastasis Bone alkaline phosphatase,

N-telopeptideHigh (blastic lesions), modest(lytic lesions)

PTH-rP IntermediateCardiac involvement Carcinoid BNP Intermediate

This table shows the specic biochemical markers used for each tumor and their specicity.

CgA and CgB indicates chromogranins A and B; HCG, human chorionic gonadotropin; ACTH, adrenocorticotropic hormone; ADH, anti diuretichormone; 5-HIAA, GRP, gastrin-releasing peptide; BNP, brain natriuretic peptide.

tumori neuroendocrine

Documents