Fig. 2.1
Histologic features of digestive neuroendocrine neoplasms as exemplified in the pancreas according to morphologic criteria for tumor grading. Well-differentiated neuroendocrine tumors of grade 1 exhibit organoid appearance featuring trabecular, insular or gyriform growth, mild atypia and abundant vasculature (a,b). Well-differentiated neuroendocrine tumors of grade 2 show trabecular to solid appearance with moderate cell atypia (c) and modified desmoplastic stroma (d). Poorly differentiated neuroendocrine carcinomas are characterized by confluent and solid growth with necrosis, marked atypia, large or small cells and plentiful mitoses (e,f)
Further complexity to the issue of grading abdominal neuroendocrine tumors is provided by the category of NEC. Traditionally considered a monolithic group of poorly differentiated and high-grade carcinomas with uniformly dismal prognosis and consistently hopeless therapy options [1], this category has recently been re-thought on the basis of the differential distribution of defining criteria, in particular Ki-67 LI and differentiation level. As a matter of fact, the wide range of cell differentiation (from well to poorly differentiated features) and of Ki-67 LI distribution (>20 up to virtually 100%) suggest that this tumor category is inherently heterogeneous in terms of molecular characteristics, clinical behavior and response to therapy [20, 38–40]. A 55% cut-off threshold has been proposed for Ki-67 LI to separate true NEC with small to large cell morphology, dismal prognosis and susceptibility to platinum agent chemotherapy from neoplasms bearing NET-like or large cell morphology, intermediate prognosis and lower response to platinum agent treatments according to the provisional term of G3 NET or well-differentiated NEC [20, 38–40]. This newly challenging tumor category would show response to alkylating agents and, to some extent, biological drugs [41].
Another puzzling observation regards the coexistence of higher and lower grade components in the same neoplasms of abdominal neuroendocrine neoplasms as highlighted by heterogeneous morphology, mitotic count, Ki-67 LI and molecular alterations [42], which challenges the concept of secondary NEC as an entity developing from preexisting NET as a result of tumor progression. This finding suggests a paradigm shift to the current knowledge on the pathogenesis of GEP NEN, but the same phenomenon is shared by NET arising in the thymus and the lung [43].
To ensure reproducibility of results, both parameters should be assayed on areas of highest immunostaining for the Ki-67 antigen or greatest concentration of mitoses (the so-called hot spots), possibly assessing the same tumor areas to minimize discrepancies due to imperfect collinearity of Ki-67 antigen and mitotic activity. To minimize interobserver variability, an actual area of 2 mm2 has become a standard reference in NET of the lung [44]. In the event of grading discrepancy, the criterion of the highest value of Ki-67 LI or mitotic count was advised, mostly identifying Ki-67 LI as the closest predictor of tumor behavior (a higher Ki-67 LI compared to the mitotic count is more likely than the opposite taking into account technical artifacts and greater difficulties in recognizing mitoses in small biopsy samples) [2, 45, 46]. The grading assignment was proven to be independent of morphology in providing NET and NEC characterization, inasmuch as either tumor category may show disconnection between morphology (more deceptive and challenging in small-sized material) and subsequent clinical behavior [2, 45]. This phenomenon is well known not only in the NET category revealing G1 NET and G2 NET, but even in the NEC category (realizing the so-called G3 NET or well-differentiated NEC as above detailed), whose heterogeneous composition has recently shown important clinical implications in terms of response to different therapies [41]. Necrosis does not play any role in the grading procedure of GEP tract NEN at variance with lung neuroendocrine tumors [35, 44, 47], although this parameter has been proposed to meliorate the prediction of prognosis [48]. This grading system has improved the definition and diagnosis of abdominal neuroendocrine tumors and identified more homogeneous populations of patients amenable to personalized treatments. Another advantage regards the better management of stage IV patients, who can be treated according to behavioral and biological characteristics of tumors rather than tumor extent, thereby avoiding overtreatment of lower tumor grade patients [16].
Potential sources of discrepancy could arise from differences in Ki-67 LI and mitotic count between primary and secondary lesions, either synchronous or metachronous, inside the same tumor mass or in different metastatic foci due to clonal selection or epigenetic changes [17]. Furthermore, different methods for assaying Ki-67 LI, either manual or automated, the use of biopsy or cytology samples vs. surgical specimens and interobserver variation of defining criteria may further account for discrepancy in tumor grade definition [33, 49]. Several tools have been proposed to minimize these drawbacks, such as multi-parametric definition combining necrosis, mitoses and Ki-67 LI, additional biopsies in case of large size tumors and/or multifocal metastasis, and the use of instrumental guidance reflecting metabolic or cell differentiation features (for example, FDG-PET in NEC or 68Gallium-PET in NET) [16].
All discrepancies should always be commented on pathology reports to provide a plausible and meaningful interpretation to tumor behavior for the clinical handling of patients [50]. As a matter of fact, the basic question is not whether to grade or not to grade abdominal neuroendocrine neoplasms but rather how to reliably grade them in the decision-making process within individual tumor patients [16]. The clinical compliance of this grading system in abdominal neuroendocrine tumors is further endorsed by its inclusion into the International Union for Cancer Control (UICC), the WHO 2010 on digestive tract tumors, and the American Joint Cancer Committee (AJCC) classification schemes and the European Neuroendocrine Tumor Society (ENETS) guidelines.
2.3 2.3 Staging
Tumor stage is also fundamental information, which never should be missing in the pathology report of abdominal neuroendocrine tumors, because it complements the prognostic information and the clinical handling of these patients. The assessment schemes on neuroendocrine tumor extent are anatomical site-specific for the definition of tumor (T) parameters based on size and invasion because of the significant differences in clinical behavior of these tumors according to the organs of origin. By contrast, for the nodal (N) and distant metastasis (M) definition, current clinical experience and biological knowledge about neuroendocrine neoplasms is insufficient for further subdividing them, beyond a mere dichotomous separation based on the occurrence or absence of tumor deposits. The current staging systems of abdominal neuroendocrine neoplasms are in keeping with the 8th edition of the UICC and AJCC classification for the single organs (stomach, duodenum/ampulla/proximal jejunum, pancreas, lower jejunum and ileum, appendix, large bowel and rectum). The ENETS staging system overlaps largely with these proposals, with some differences and conflicting results in the prognostic stratification of neoplasms arising in the stomach, pancreas and appendix [26, 27]. In cases of discrepancy, both staging classifications should be mentioned and the worst stage used for classification [35]. For example, the ENETS staging system of pancreatic NEN is better suited than the UICC and AJCC proposal to forecasting patient survival, while the opposite holds true for appendix NEN. Lastly, in the stomach, both the TNM/AJCC and ENETS systems predict survival especially in NEC, while the TNM/AJCC seems to be superior in the NET subgroups [51]. An integration of staging and grading (the so-called TGM system) has been proposed in keeping with other malignancies as a tool for better predicting survival in pancreatic NEN as compared with singly used stage and grade schemes [52], but further confirmation is still needed before accepting this innovative approach into the clinical practice of abdominal neuroendocrine neoplasms.
2.4 2.4 Conclusive Remarks
Pathology reports of abdominal neuroendocrine tumors should always contain information useful for the best clinical handling of patients. In particular, anatomical location, appropriate classification (tumor vs. carcinoma), differentiation levels (well vs. poor differentiated), 2010 WHO/ENETS tumor grading (G1 to G3) with details on mitotic count and Ki-67 LI, and the TNM/AJCC/ENETS pathological stage should not be missing in any diagnosis of abdominal neuroendocrine neoplasms, especially resection specimens. Small biopsy samples in turn are most often the only available material in the setting of metastatic NEN, where grading assessment is clinically warranted. The time-honored and conceptually surpassed term “carcinoid” should be avoided, and only confined to carcinoid syndrome-associated NET. On surgical specimens, additional histologic features such as microscopic multicentric disease, presence and extent of non-ischemic necrosis, vascular and perineural invasion, definition of resection margins and evaluation of the functional status may help better complement the overall clinical assessment. Upon request or inside specific trials, information about predictive markers, such as somatostatin receptors, m-TOR pathway molecules or thymidylate synthase expression, may be integrated into the pathology reports to best manage these patients on clinical grounds. All pathology diagnoses should be discussed within multidisciplinary teams.
References
1.
Bosman F, Carneiro F, Hruban R, Theise ND (2010) WHO classification of tumours of the digestive system, 4th edn. IARC Press, Lyon
2.
3.
4.
Falkmer S (1993) Phylogeny and ontogeny of the neuroendocrine cells of the gastrointestinal tract. Endocrinol Metab Clin North Am 22(4):731–752PubMed
5.
Rindi G, Grant SG, Yiangou Y et al (1990) Development of neuroendocrine tumors in the gastrointestinal tract of transgenic mice. Heterogeneity of hormone expression. Am J Pathol 136(6): 1349–1363PubMed
6.
Schonhoff SE, Giel-Moloney M, Leiter AB (2004) Minireview: development and differentiation of gut endocrine cells. Endocrinology 145(6):2639–2644CrossrefPubMed