benign endometrium: dysfunctional bleeding, breakdown
Transcription
benign endometrium: dysfunctional bleeding, breakdown
BENIGN ENDOMETRIUM: DYSFUNCTIONAL BLEEDING, BREAKDOWN, AND METAPLASIA Michael T. Mazur, M.D. ClearPath Diagnostics 600 E. Genesee St. Syracuse, NY 13066 [email protected] Abnormal uterine bleeding (AUB) is a common sign of a number of different uterine disorders ranging from dysfunctional (nonorganic) abnormalities or complications of pregnancy to organic lesions such as polyps, hyperplasia, or carcinoma.(1-7) In many cases, AUB leads to endometrial biopsy or curettage that can present unique diagnostic challenges for pathologists. There are a variety of terms are applied to AUB. Dysfunctional uterine bleeding (DUB) refers to bleeding due to ovulatory dysfunction with no other uterine or systemic abnormality present. Post-menopausal bleeding (PMB) describes uterine bleeding one year after the cessation of menses. By definition, DUB excludes postmenopausal bleeding or bleeding due to the presence of specific pathologic processes such as inflammation, polyps, hyperplasia, carcinoma, exogenous hormone effects, and complications of pregnancy. The endometrial changes associated with DUB are important to recognize, because they may be confused with more serious lesions such as hyperplasia. A frequent concern, especially in the perimenopausal and postmenopausal patient, is the presence of hyperplasia or carcinoma of the endometrium. Overt hyperplasia or neoplasia often is the least troublesome problem from a diagnostic point of view. A variety of other, benign patterns are more commonly seen and present some of the greatest challenges in biopsy interpretation since they are difficult to catalog. Artifacts, superimposed patterns of breakdown and bleeding, ovulatory disorders, hormonal effects, and metaplasia all contribute to the complexities of interpreting these cases. 1 DYSFUNCTIONAL UTERINE BLEEDING Dysfunctional uterine bleeding is a clinical term that refers to bleeding due to irregularities in ovarian function, so it is most common at the time of menopause. DUB is largely a diagnosis of exclusion once other etiologies such as organic lesions, endometritis and systemic bleeding disorders are ruled out.(8-12) Endometrial biopsy or curettage is usually done to determine if endometrial changes are consistent with DUB and to exclude other causes of bleeding, especially organic lesions in the perimenopausal woman. These biopsies are common in many practices, usually showing benign changes that may be complicated by breakdown and bleeding and by abnormal endometrial development that is neither hyperplastic nor neoplastic. DUB-associated patterns have the stigma of lacking glamour of a more rare and significant diagnosis such as atypical hyperplasia. Despite their somewhat banal nature, they can be complicated to interpret and may require considerable study. The terminology applied to the consequences of dysfunctional patterns also has not been the target of strict conventions of nomenclature, probably allowing many different terms for groups of fundamentally similar changes. DUB can be due to anovulatory cycles or to disorders in follicle development during the luteal phase. In most cases, DUB is due to anovulatory cycles, and some authors regard anovulation as the only cause. Alternatively, however, a shortened or prolonged luteal phase may also lead to abnormal bleeding, concepts known as “luteal phase defect” and “irregular shedding.” Anovulatory bleeding pattern (proliferative with glandular and stromal breakdown). Most cases of DUB are due to anovulatory cycles, a sporadic but common occurrence, especially in perimenarchal and perimenopausal women.(2) Anovulatory cycles are also a component of polycystic ovarian disease or Stein-Leventhal syndrome with more persistent endocrine imbalance in the reproductive years. In the normal menstrual cycle a cohort of follicles is recruited and they begin to develop and produce estradiol. A dominant follicle then ruptures following the LH surge at mid-cycle. After the follicle ruptures, a corpus luteum develops, producing progesterone as well as estradiol. 2 In anovulatory cycles the follicles are recruited but there is no ovulation, usually due to disorders at the hypothalamus/pituitary level or feedback signals. As a consequence, there is a surge of estradiol without progesterone from a corpus luteum. The follicle or follicles can either involute leading to estrogen withdrawal or persist leading to sustained estrogen stimulation of the endometrium. In either event the endometrium proliferates without a normal luteal phase. With estrogen withdrawal, breakdown occurs in a weakly proliferative background as the estrogen stimulus wanes (estrogen withdrawal bleeding). With estrogen persistence, the endometrium continues to proliferate with thrombi forming in superficial vessels leading to areas of breakdown (estrogen breakthrough bleeding). In either case the breakdown often affects only a portion of the endometrium. The usual morphologic reflection of anovulatory cycles is a proliferative phase pattern, often with fibrin thrombi in small vessels and breakdown and bleeding superimposed. If active bleeding is not taking place at the time of biopsy, then the changes are simply those of proliferative phase patterns. These common histologic findings are important to observe since they provide useful information for the gynecologist. Tissue fragmentation and artifacts along with the breakdown patterns often distort the tissue, but the anovulatory bleeding pattern usually is recognizable. Disordered proliferative phase. The disordered proliferative phase pattern usually is an extension of anovulatory cycles due to persistent estrogen stimulation. In this situation the endometrium is proliferative but shows focal gland irregularities including dilatation and branching like that seen in hyperplasia. In contrast to hyperplasia, however, gland irregularities are mild and focal in the disordered proliferative phase pattern. The diagnosis of disordered proliferative phase pattern has become commonplace in many practices. This term is often overused and misapplied, however. Changes such as fragmentation, telescoping and even portions of basalis can be misclassified as disordered proliferative phase patterns. In addition, the distinction between disordered proliferative patterns and simple hyperplasia is blurred in some situations. To be clinically useful this diagnosis should be reserved for those cases that truly show focal irregularities in proliferative gland 3 development. Besides anovulatory cycles, limited sampling of a polyp may yield a pattern of disordered glands. DUB and secretory changes. DUB may also develop when ovulation occurs but the corpus luteum does not develop and persist for a normal duration over the last half of the menstrual cycle. Disturbances in the rate and amount of progesterone produced by the granulosa cells of the corpus luteum result in alterations in the pattern of secretory phase development. These changes may be due to insufficient development or persistence of the corpus luteum (luteal phase defect, LPD) or to abnormal persistence of a corpus luteum (irregular shedding). (13-16) Luteal phase defect and irregular shedding are less well-defined entities. These conditions, if they occur, are sporadic and not amenable to detailed clinical-pathologic correlations to clearly define the morphologic changes. There are no well-defined morphologic changes that are diagnostic of disturbed corpus luteum function. There are clear situations, however, where there is abnormal secretory phase maturation with or without superimposed nonmenstrual breakdown. The term “irregular maturation” describes secretory endometrium that does not show a normal and universal pattern of secretory development that allows clear histologic dating according to established criteria. The endometrium can show marked variation in secretory development from area to area with some glands demonstrating tortuosity and secretions while other glands are underdeveloped, lacking these changes. It is not clear that these patterns truly reflect an inadequate corpus luteum, but the descriptive diagnosis serves to indicate that abnormal but benign secretory changes are present and clinical correlation is needed. When bleeding is present in a secretory but non-menstrual background, a descriptive diagnosis of “secretory bleeding pattern” with a brief comment serves to communicate the changes to the clinician. Rarely irregular shedding may result in a mixed phase pattern due to abnormal persistence of the corpus luteum. In this case portions of the endometrium are secretory and others areas have proliferative features. Recognition of abnormal secretory phase patterns is important. The best method to detect these changes is to recognize when secretory endometrium does not show a consistent pattern of 4 normal “datable” changes. Usually, it is not possible to determine the etiology, and there are a number of possible causes of abnormal secretory changes in addition to ovulatory dysfunction (Table 1). Consequently, a descriptive diagnosis without assigning an underlying cause is the best approach for the pathologist. BREAKDOWN PATTERNS AND METAPLASIA Glandular and stromal breakdown. When endometrium undergoes acute nonphysiologic (non-menstrual) breakdown and bleeding, glandular and stromal changes occur that are almost unique to this tissue.(1) Fibrin thrombi form in small arteries and capillaries or in dilated superficial venules resulting in apoptosis with nuclear debris at the base of glands and within the stroma.(17-19) The devitalized tissue then demonstrates a characteristic pattern of collapse as the stromal cells condense into tight clusters with scant cytoplasm and hyperchromatic, closely apposed nuclei. As these clusters of stroma ("blue balls") separate from underlying intact endometrium, they often retain a cap of epithelial cells, resulting in small round to polypoid tissue fragments. The breakdown pattern is highly variable, ranging from limited foci to diffuse changes with extensive fragmentation of the tissue. As bleeding continues and becomes chronic the endometrium may show other features including hemosiderin deposition, foam cells, and stromal hyalinization (Table 2). Another change that reflects breakdown and bleeding is eosinophilic syncytial change (ESC).(20-22) This alteration, also termed “papillary syncytial change,” occurs along the surface epithelium, occasionally extending superficially into glands. Other features of active bleeding such as stromal collapse are almost always present in close proximity or subjacent to ESC. Previously this phenomenon was termed “papillary syncytial metaplasia,”(23) but it is more degenerative than metaplastic. ESC shows no to minimal proliferative activity.(24) ESC is characterized by aggregates of stratified eosinophilic cells, often forming small papillary-like tufts lacking a connective tissue core. The syncytial aggregation of pink epithelial cells over clusters on condensed stroma contributes to the pseudo-papillary arrangements. The 5 cells of ESC are generally oval, and their nuclei have a random, haphazard distribution. The cytoplasm is pale to eosinophilic with occasional vacuoles. Cell borders are indistinct. Overall, the nuclei are cytologically bland, but some cases of ESC show mild nuclear atypia manifested by slight hyperchromasia, pleomorphism, and irregular nuclear outlines. ESC consistently has associated cellular necrotic debris and often has a neutrophilic infiltrate, too. This abnormality can be confused with metaplasia, atypia, or neoplasia. In contrast to metaplastic or neoplastic lesions that largely involve glands, however, ESC is usually limited to surface epithelium. Metaplasia. Endometrial epithelium can show a variety of cytoplasmic changes commonly termed “metaplasia.” These epithelial changes frequently occur in endometria that contain hyperplasia or neoplasia, but they can occur in a variety of other benign conditions, especially polyps. Many disorders previously classified as metaplasia are better termed "change" since they are not true metaplastic transformations of the epithelium.(1) There are five general types of cytoplasmic change seen in the endometrium.(25) These are squamous, ciliated cell (tubal), eosinophilic, mucinous and secretory (clear cell) change. Eosinophilic cell change often is cytologically related to tubal metaplasia but lacks cilia. Association of eosinophilic cell change with mucinous metaplasia in some cases suggests a relation between the two cell types with eosinophilic change representing a subtype of immature mucinous metaplasia.(26) Tubal metaplasia usually shows immunoreactivity for p16 as well as aberrant expression of some cell cycle proteins, suggesting it has potential to be a premalignant lesion.(27) Progestin therapy or hyperplasia and well-differentiated adenocarcinoma can further enhance various patterns of cytoplasmic change or metaplasia.(28;29) Eosinophilic syncytial change, discussed above along with other features of breakdown and bleeding, is not a metaplasia and is unrelated to the other forms of cytoplasmic transformation. Like some changes in breakdown such as artifactual crowding and ESC, metaplastic patterns complicate and confound the interpretation of specimens. Since metaplasia frequently accompanies hyperplasia or well-differentiated carcinoma, separating benign metaplastic changes from atypical changes is of paramount importance. One issue with metaplasia is that these changes, except for secretory change and endocervical-type mucinous change, result in cytoplasmic eosinophilia, a feature shared with gland cells in atypical hyperplasia and low grade 6 adenocarcinoma as well as the unrelated phenomenon of ESC. The cytologic details of cells in question are the key to making the critical distinction. Metaplastic change should show no atypia. The cells may be pseudostratified in metaplasia, but orientation to the basement membrane is maintained in contrast to atypical epithelium that shows loss of polarity in relation to the underlying basement membrane. The nuclei of metaplasia lack atypia, a feature of atypical hyperplasia (Table 3) and many well differentiated adenocarcinomas. Squamous change presents a different challenge. Squamous change complicates gland patterns because the process can result in crowding of the glands secondary to distention by nests of non-keratinizing squamous cells. In this situation, the other histologic features including cellular atypia of the glandular component and, in the case of carcinoma, confluent gland arrangements, determine the correct diagnosis. TERMINOLOGY For abnormal endometrium that lacks a specific organic abnormality, selecting the best diagnostic term can be as challenging as the interpretation of the specimen. The gynecologist wishes to know the following: 1) Is there an organic lesion such as a complication of pregnancy, inflammation or a polyp? 2) Is there evidence of active or old breakdown and bleeding? 3) Is there evidence to suggest dysfunctional bleeding? 4) Is there evidence of hyperplasia, atypia, or carcinoma? When a biopsy is done for DUB, the report should address the presence or absence of morphologic changes of breakdown and bleeding as well as any specific lesions. If the pattern is that of proliferative endometrium with breakdown and if the clinical history is appropriate, the changes can be accurately attributed to anovulatory cycles. A descriptive diagnosis such as “proliferative endometrium with glandular and stromal breakdown” offers a clear morphologic 7 interpretation of the bleeding pattern that often is sufficient for clinical management. An additional comment indicating that the change is compatible with anovulatory cycles helps to clarify the diagnosis. If the changes show non-menstrual secretory endometrium with breakdown but these are not diagnostic of a defined eiomy phase abnormality, descriptive terms such as “secretory bleeding pattern” communicates the observation of an abnormal yet benign appearance while not assigning definite morphologic etiology. In general a comment regarding the absence of other possible causes of bleeding such as hyperplasia, inflammation, pregnancy, or polyps is most useful in addressing specific clinical concerns. Occasional biopsies show extensive breakdown and bleeding that largely obscures the cytologic details of the glands and stroma. Although it is usually possible to exclude neoplastic processes in such cases, detailed assessment of the endometrium to determine the underlying pathologic process becomes difficult. Unless the breakdown is clearly menstrual, i.e. reflecting the shedding at the end of a normal ovulatory cycle, breakdown patterns should not be diagnosed as “menstrual.” Instead, it is better to use descriptive diagnoses that reflect the morphologic changes. Descriptive diagnoses should be used carefully, however. The term “dyssynchronous” endometrium has been used to describe apparent alterations in secretory phase development yet this term does not have a specific connotation or meaning. Its use can be confusing unless there is clear communication between the pathologist and gynecologist regarding its meaning, and I don’t use it. Likewise, the terms “withdrawal” and “breakthrough” should be avoided in pathologic diagnoses because they lack clear definitions in the clinical literature regarding endometrium bleeding. It is best to avoid other vague terms such as “lytic endometrium.” In summary, the endometrial biopsy showing benign, non-hyperplastic changes presents a distinct group of challenges for the pathologist. A myriad of normal and pathologic processes with varied histology may be encountered. In the absence specific lesions such as endometritis, polyp or hyperplasia, a descriptive evaluation that clearly describes the changes present will help guide the gynecologist in patient management. 8 TABLE 1 POSSIBLE CAUSES OF ABNORMAL SECRETORY PHASE PATTERNS Luteal phase defects Persistent corpus luteum (irregular shedding) Organic lesions (polyps, secretory hyperplasia, etc.) Submucosal leiomyomas Intrauterine adhesions Inflammation Complications of pregnancy Progestin effects TABLE 2 HISTOLOGIC FEATURES OF BREAKDOWN AND BLEEDING Stromal “collapse” with cell clusters Eosinophilic syncytial change Fibrin thrombi Nuclear debris at base of gland cells Nuclear debris in stroma Hemosiderin Foam cells Stromal fibrosis and hyalinization 9 TABLE 3 FEATURES OF ENDOMETRIAL EPITHELIAL ATYPIA Nuclei enlarged, irregular Loss of nuclear polarity Chromatin clumping (vesicular appearance) Prominent nucleoli Cytoplasmic eosinophilia, diffuse or focal 10 Reference List (1) Mazur MT, Kurman RJ. Diagnosis of endometrial biopsies and curettings. A practical approach. New York: Springer Science+Business Media, 2005. (2) Speroff L, Glass RH, Kase NG. Clinical gynecologic endocrinology and infertility. 6 ed. Baltimore: Lippincott Williams & Wilkins, 1999. (3) Butler WJ. Normal and abnormal uterine bleeding. In: Rock JA, Jones HW, III, editors. Te Linde's operative gynecology. Philadelphia: Lippincott Williams and Wilkins, 2003: 457-481. (4) Galle PC, McRae MA. Abnormal uterine bleeding. Finding and treating the cause. Postgrad Med 1993; 93:73-81. (5) Kilbourn CL, Richards CS. Abnormal uterine bleeding. Diagnostic considerations, management options. Postgrad Med 2001; 109(1):137-4, 147. (6) Stenchever MA. Differential diagnosis of major gynecologic problems by age groups. In: Stenchever MA, Droegemueller W, Herbst AL, Mishell DR, Jr., editors. Comprehensive gynecology. St. Louis: Mosby, Inc., 2001: 155-177. (7) Wren BG. Dysfunctional uterine bleeding. Aust Fam Physician 1998; 27(5):371-377. (8) Aksel S, Jones GS. Etiology and treatment of dysfunctional uterine bleeding. J Obstet Gynecol 1974; 44:1-13. (9) Altchek A. Dysfunctional uterine bleeding in adolescence. Clin Obstet Gynecol 1977; 20:633-650. (10) Bayer SR, DeCherney AH. Clinical manifestations and treatment of dysfunctional uterine bleeding. JAMA 1993; 269:1823-1828. (11) Scommegna A, Dmowski WP. Dysfunctional uterine bleeding. Clin Obstet Gynecol 1973; 16:221-254. (12) Vakiani M, Mawad J, Talerman A. Heterologous sarcomas of the uterus. Int J Gynecol Pathol 1982; 1:211-219. (13) Buckley CH, Fox H. Biopsy pathology of the endometrium. London: Arnold, 2002. (14) Dallenbach-Hellweg G. Histopathology of the endometrium. 4 ed. New York: SpringerVerlag, 1987. (15) Sherman ME, Mazur MT, Kurman RJ. Benign diseases of the endometrium. In: Kurman RJ, editor. Blaustein's pathology of the female genital tract. New York: Springer-Verlag, 2002: 421-466. (16) Vakiani M, Vavilis D, Agorastos T, Stamatopoulos P, Assimaki A, Bontis J. Histopathological findings of the endometrium in patients with dysfunctional uterine bleeding. Clin Exp Obstet Gynecol 1996; 23(4):236-239. (17) Ferenczy A. Pathophysiology of endometrial bleeding. Maturitas 2003; 45(1):1-14. (18) Picoff RC, Luginbuhl WH. Fibrin in the endometrial stroma: Its relation to uterine bleeding. Am J Obstet Gynecol 1964; 88:642-646. (19) Stewart CJ, Campbell-Brown M, Critchley HO, Farquharson MA. Endometrial apoptosis in patients with dysfunctional uterine bleeding. Histopathology 1999; 34(2):99-105. 11 (20) Silverberg SG, Kurman RJ. Tumors of the uterine corpus and gestational trophoblastic disease. Atlas of tumor pathology, 3rd series, Fascicle 3. Washington, DC: Armed Forces Institute of Pathology, 1992. (21) Zaman SS, Mazur MT. Endometrial papillary syncytial change. A nonspecific alteration associated with active breakdown. Am J Clin Pathol 1993; 99:741-745. (22) Norimatsu Y, Shimizu K, Kobayashi TK, Moriya T, Kaku T, Tsukayama C et al. Endometrial glandular and stromal breakdown, part 2: cytomorphology of papillary metaplastic changes. Diagn Cytopathol 2006; 34(10):665-669. (23) Clement PB. Pathology of the uterine corpus. Hum Pathol 1991; 22:776-791. (24) Shah SS, Mazur MT. Endometrial eosinophilic syncytial change related to breakdown. Immunohistochemical evidence suggests a regressive process. In press . 2007. (25) Hendrickson MR, Kempson RL. Endometrial epithelial metaplasias: proliferations frequently misdiagnosed as adenocarcinoma. Report of 89 cases and proposed classification. Am J Surg Pathol 1980; 4:525-542. (26) Moritani S, Kushima R, Ichihara S, Okabe H, Hattori T, Kobayashi TK et al. Eosinophilic cell change of the endometrium: a possible relationship to mucinous differentiation. Mod Pathol 2005; 18(9):1243-1248. (27) Horree N, Heintz AP, Sie-Go DM, van Diest PJ. p16 is consistently expressed in endometrial tubal metaplasia. Cell Oncol 2007; 29(1):37-45. (28) Miranda MC, Mazur MT. Endometrial squamous metaplasia. An unusual response to progestin therapy of hyperplasia. Arch Pathol Lab Med 1995; 119(5):458-460. (29) Wheeler DT, Bristow RE, Kurman RJ. Histologic alterations in endometrial hyperplasia and well-differentiated carcinoma treated with progestins. Am J Surg Pathol 2007; 31(7):988-998. 12 BENIGN ENDOMETRIUM: DYSFUNCTIONAL BLEEDING, BREAKDOWN, AND METAPLASIA Michael T. Mazur, M.D. ClearPath Diagnostics Syracuse, NY [email protected] AUB: Most Common • Dysfunctional bleeding • Polyps • Atrophy Abnormal Uterine Bleeding: Sentinel of Endometrial Pathology • Dysfunctional • Organic lesions – Polyps, Hyperplasia, Carcinoma, etc. • • • • • Atrophy Exogenous hormones Complications of pregnancy Inflammation Systemic bleeding disorders Dysfunctional Uterine Bleeding • Abnormal bleeding with no organic cause • Pathogenesis: A. Anovulatory cycles B. Luteal phase abnormalities 1. Luteal phase defect 2. Irregular shedding Anovulatory Cycles Anovulatory Bleeding • Follicles develop, but no ovulation with rupture • Estradiol production leads to endometrial proliferation • Variable atresia or persistence of follicle • Follicles involute No estrogen, withdrawal bleeding • Follicles persist Estrogen production, vascular ectasia, breakthrough bleeding Anovulatory Bleeding Pattern • Proliferative phase glands and stroma, variable amount • Glands may have slight disorganization (disordered) • Glandular and stromal breakdown, focal to diffuse Proliferative with partial breakdown Fibrin thrombi and stromal collapse Proliferative with focal breakdown Anovulatory bleeding pattern with marked fragmentation Disordered Proliferative Phase Pattern • Mildly irregular gland shapes and sizes • May be result of anovulatory cycles • Diagnosis often over-used Luteal Phase Defect • Inadequate progesterone production from corpus luteum, possibly due to premature regression • Associated with infertility • Role in DUB poorly defined Histologic Features Of LPD • Histologic date lags by more than 2 days, or possibly • Irregular maturation, or • Non-menstrual secretory with breakdown Secretory with irregular maturation Secretory with irregular maturation Secretory bleeding pattern Causes of Abnormal Secretory Phase Development • Luteal phase abnormalities • Organic lesions – Polyps, secretory hyperplasia, etc. • • • • Leiomyomas Adhesions Chronic inflammation Progestin effects Secretory bleeding pattern Non-menstrual Breakdown (Bleeding Pattern) • Pattern of early necrosis of the endometrium • Specific and unique morphologic features • Changes along with fragmentation can mimic other lesions Breakdown with artifactual crowding Breakdown And Bleeding • • • • • Stromal “collapse” with clusters Nuclear debris (apoptosis) Fibrin thrombi Eosinophilic syncytial change Hemosiderin, foam cells, hyalinized stroma Complex hyperplasia Eosinophilic Syncytial Change Eosinophilic Syncytial Change • Syncytial aggregates of pink epithelium • Usually along surface epithelium • Cytologically bland • Can form pseudo-papillary tufts • A marker of endometrial breakdown in a variety of conditions, many not hyperplastic or neoplastic • Regressive change unrelated to “metaplasia” Focal breakdown with ESC and stromal clusters Anovulatory bleeding pattern with ESC and stromal clusters Proliferation Comparison Ki-67 pHH3 ESC 1.3% 0 AH 15.8% 2.3% Gr 1 CA 10.6% 2.4% Ki-67: MIB-1 antibody pHH3 = phospho-histone H2 Ser 28 (mitosis marker) Extensive breakdown with ESC and pseudo-papillary pattern Endometrial Cytoplasmic Changes (Metaplasia) • • • • • Squamous Ciliated cell (tubal) Eosinophilic (pink cell) Mucinous Secretory (clear cell) Eosinophilic (Pink) Epithelial Cells • • • • • • • Atypical hyperplasia Adenocarcinoma Squamous differentiation Ciliated cell change Eosinophilic change Mucinous change Eosinophilic syncytial change (ESC) Endometrial Cytoplasmic Change (Metaplasia) • Usually an estrogenic effect • May be due to irritation or trauma, e.g., polyp or inflammation • Often associated with hyperplasia or carcinoma; amplified with progestin therapy Atypical Hyperplasia Diagnostic Features • Cytologic atypia, focal or diffuse. Irregular nuclei with chromatin clumping and prominent nucleoli • Loss of nuclear polarity • Cytoplasm abundant with dense eosinophilia, focal or diffuse Ciliated/Eosinophilic Cell Change • Nuclei often round, stratified • Smaller, uniform nuclei without chromatin changes, nucleoli of atypical cells • Luminal border usually sharp Atypical hyperplasia Ciliated cell change in simple hyperplasia Eosinophilic cell change in benign polyp Mucinous Change • Variable presentation: from simple endocervical-type epithelium to complex patterns • Eosinophilic change related to mucinous change in some cases. Mucinous change in hyperplasia without atypia Eosinophilic and mucinous change in hyperplasia Squamous Change (Squamous Differentiation) • Seen in polyps, hyperplasia, carcinoma, inflammation • Often non-keratinizing • Luminal clusters (morules) gives crowded gland pattern Benign polyp with squamous metaplasia Complex atypical hyperplasia with squamous change Complex atypical hyperplasia with squamous change Well-differentiated adenocarcinoma with squamous change Metaplasia (Cytoplasmic Change) Common Concerns • Occur in a variety of conditions, often hyperplasia or polyp • ESC is not related to “metaplasia” • Evaluate cytologic detail to separate from significant atypia • Is the specimen representative and adequate? • Is hyperplasia, atypia or carcinoma present? • Abnormal, but benign - what is the best diagnosis? The Report: Abnormal, Benign and Difficult to Classify • Exclude other pathology, e.g., polyps, hyperplasia, atypia, etc. • Minimize comments about metaplasia/cytoplasmic change • Diagnose descriptively Descriptive Diagnoses, Examples • Proliferative with glandular and stromal breakdown • Abnormal secretory bleeding pattern • Secretory with irregular maturation (Avoid: “breakthrough,” “withdrawal,” “dyssynchronous,” “lytic”, etc.) ENDOMETRIAL INTRAEPITHELIAL NEOPLASIA George L. Mutter MD Associate Professor of Pathology, Harvard Medical School Div. of Women's and Perinatal Pathology, Department of Pathology Brigham and Women's Hospital, 75 Francis Street, Boston, MA 02115 website: www.endometrium.org Biology of Endometrial Intraepithelial Neoplasia 1 Endometria Intraepithelial Neoplasia (EIN) is a clonal proliferation of architecturally and cytologically altered premalignant endometrial glands which are prone to malignant transformation to endometrioid (Type I) endometrial adenocarcinoma. EIN lesions are non-invasive genetically altered neoplasms which arise focally, and may convert to malignant phenotype upon acquisition of additional genetic damage. Diagnostic criteria for EIN have been developed by histopathologic correlation with clinical outcomes, molecular changes, and objective computerized histomorphometry. EIN should not be confused with unrelated serous intraepithelial carcinoma (serous EIC), which is an early phase of (Type II) papillary serous adenocarcinomas of the endometrium. Management of EIN lesions follows guidelines long established for atypical endometrial hyperplasia. A high concurrent cancer rate (26%), and concern that sampling errors may miss an occult tumor, have led to a prevailing view that immediate hysterectomy is justified by its combined diagnostic and therapeutic benefits. Young patients wishing to preserve fertility, and women who are poor surgical risks, are candidates for hormonal (progestin) therapy. Systemic progestins can successfully ablate up to 90% of endometrial precancers in young women 2, although it is not possible in advance to predict that fraction which will respond. A decision to treat hormonally must thus be made between the clinician and patient in full light of the risks, and with the precondition that regular followup surveillance can be performed. Figure 3: Clonal Origin of EIN. The first genetic changes (such as PTEN inactivation) which Malignant Transformation Initiation Normal Histology EIN Adenocarcinoma (Polyclonal ? Latent “Clone”) (Monoclonal Premalignant Neoplasm) (Monoclonal Malignant Neoplasm) initiate endometrial carcinogenesis are unaccompanied by any phenotypic alterations at the light microscopic level. This “latent”, phase of cytologically and architecturally normal but genetically altered cells may persist for years in a normally menstruating woman. Low cancer risk, combined with lack of a rational therapeutic response, are reasons that systematic screening and treatment of these “latent” phase lesions is unwarranted at present. As additional genetic damage accumulates, higher risk morphologically altered mutant clones declare themselves by demonstrating those architectural and cytologic alterations that distinguish EIN. Malignant transformation of EIN lesions, which occurs at least 46-times more frequently than non-EIN tissues, warrants careful diagnosis and treatment. Endocrine modifiers of endometrial cancer risk act upon the latent and EIN phases of this sequence by tipping the balance of clonal expansion vs. involution. A combined molecular and histopathologic model for EIN: Latent, premalignant, and malignant phases of EIN-mediated endometrial carcinogenesis are diagrammed in Figure 3. In almost half of apparently normal women, histologically unremarkable proliferative endometria contain a small fraction of (PTEN tumor suppressor gene) mutant endometrial glands. This phase may be construed as “latent” because not only do the mutated glands look completely normal under the microscope, but they progress to EIN and cancer at very low efficiency. This latent phase may persist for years, with continued presence of scattered and interspersed mutant glands after many menstrual cycles 3. Mutant glands are probably represented in the reserve population of cells that regenerate a new functionalis each month. Endocrine factors act upon these “latent precancers” to modulate involution, or progression to EIN. Transition to EIN requires accumulation of additional genetic damage in at least one “latent precancer” cell, which then clonally expands from its point of origin (indicated by expanding arrows) to form a contiguous grouping of a tightly packed and cytologically altered glands recognizable as EIN. The monoclonal precancer (EIN) develops internal heterogeneity through mutation, and advantageous events selected by local conditions result in hierarchical subclones (left to right) of varying success. EIN lesions have only marginal increases in growth potential, and retain susceptibility to further growth modulation by hormonal factors. Some involute. Others, through additional mutation and selection, reach a stage where hormonal support is no longer required for survival. Malignant transformation to cancer is defined by accumulation of sufficient genetic damage to permit invasion of adjacent stromal tissues. 1.What Is EIN? Endometrial Intraepithelial Neoplasia, EIN 4;5, is the histopathologic presentation of premalignant endometrial disease which confers an elevated risk for endometrial cancer. The singular category of EIN is not stratified or divided into subgroups, and must be distinguished from earlier phases of latent premalignant disease, and endometrial carcinoma. This term was proposed by The Endometrial Collaborative Group 4 to accommodate changing concepts of premalignant endometrial disease and take advantage of revised diagnostic strategies. EIN needs to be treated, and the type of therapy decided between the patient and treating physician. Things that may influence the choice of surgical vs. hormonal therapy include but are not limited to: diagnostic confidence that a co-existing carcinoma has been excluded, desire for maintained fertility, ability to perform followup surveillance, and patient-specific hormonal and surgical risks. 2.Clinicopathologic Foundations Of EIN Rigorous experimental validation of clinically and biologically defined endometrial precancers, and development of correlative diagnostic criteria is a multidisciplinary process. Key predictions expected of precancers 6 which have now been fulfilled for EIN, and practical aspects of their clinical implementation are listed in Table III: Table III: Precancer postulates fulfilled for EIN Postulate Precancers differ from normal tissues Precancers share some, but not all features with carcinoma Precancers can be diagnosed Precancers increase risk for carcinoma Epidemiologic and genetic mechanisms are linked Introducing precancer genotype into an animal produces premalignant lesions and heightened cancer risk Evidence Monoclonal 7-9. Divergent genotype 10. Including PTEN 11-13, K-ras 14-16, and MLH1 changes 17. Both are monoclonal 7-9;18. Precancer-cancer lineage hierarchy 10. Computerized morphometry reference standard for EIN 18 High concurrent cancer rate in EIN 19;20;20 High future cancer rate in EIN 21-24 The PTEN gene, mutated in EIN, is subject to hormonal modulation 13;25 100% of PTEN mutant heterozygote mice get endometrial “hyperplasia” and 21% evolve to carcinoma. 26 3.WHO Hyperplasia-EIN Concordances Concordances with EIN diagnostic system and were obtained by review of cases initially diagnosed using other endometrial hyperplasia Complex Simple classification schemes 21. Atypical Non-Atypical Non-Atypical Figure 4: Correlation of WHO and EIN Diagnoses. Gray portions of Bar Graphs show approximate percentages of each WHO hyperplasia class that will be diagnosed as EIN. Remaining WHO hyperplasias not diagnostic of EIN (white) will be allocated to unopposed estrogen (anovulatory), polyp, and other categories. Pie chart shows relative contributions of each hyperplasia type to the EIN diagnostic category in a series of 97 cases with 28 EIN examples 21. Hyperplasia Hyperplasia 78% 44% Hyperplasia 4% 29% 7% 4.Clinical Cancer Outcomes Following EIN Diagnosis 64% The risk of developing endometrial cancer, as predicted by an EIN diagnosis are the basis for therapy. Endometrial Intraepithelial Neoplasia Although there are many previous references citing cancer outcomes of EIN patients 19;22;23, the two studies summarized below show cancer predictive value of subjective (Figure 5) 21 and objective histomorphometric (Figures 6-8) 20 EIN diagnosis. Patients with EIN lesions have an overall 89-fold increased cancer risk than those without EIN. In practice, the time interval separating EIN from cancer divides these into either concurrent EIN and cancer, or progression events from EIN to cancer. For purposes of illustration we have considered cancers diagnosed within 12 months of EIN to be “concurrent” (Figure 7), and those following EIN by more than one year to be “progression events” (Figure 8). Figure 5: Cancer outcomes (black), by followup interval (vertical axis) of 97 endometrial biopsies diagnosed by WHO hyperplasia (left) or EIN (right) schema 21 . Endometrial hyperplasias (left panel) were rediagnosed subjectively (without morphometry) as EIN or benign, non-EIN (right panel). All 8 cancer outcomes (black symbols) followed an initial diagnosis of EIN. EIN has a better negative predictive value than atypical hyperplasia, as 2/8 cancer occurrences were seen in the nonatypical hyperplasia groups. Hyperperplasia Schema EIN Schema Followup Interval, Days 2000 1500 1000 500 0 Outcome No Cancer Cancer le x mp al Co pic aty 1.0 Cancer Free Survival Figure 6: Overall cancer free survival of 674 patients with “endometrial hyperplasia” stratified by morphometry into EIN (D-Score <1) or benign non-EIN (D-Score>1) 20. 65/67 cancer occurrences occurred in the EIN category. Elevated cancer risk of having an EIN lesions is 89 times that of women without EIN. Incidences of carcinoma following EIN diagnosis may be considered concurrent (steep part of curve in months 1-12) or future (more shallow curve > 12 months). These subsets of short and long term cancer occurrences are plotted for this dataset in Figures 3 and 4. 2/446 non-EIN and 65/228 EIN cases developed adenocarcinoma. lex ple pia mp pia Sim aty Co aty no no n, n ig I N Be n-E No N EI Benign (DS>1) 0.8 0.6 0.4 EIN (DS<1) 0.2 HR=89 0.0 0 50 100 150 Followup Time, Months 200 1.0 Benign (DS>1) 0.8 Cancer Free Survival Figure 7: Concurrent Cancer in women with EIN. “Concurrent cancers,” those diagnosed within 1 year of a baseline cancer-free endometrial biopsy, are more likely to be seen in women with EIN compared to women without EIN. Approximately half of patients with EIN lesions will have a cancer diagnosed in the first year. 197 Women with “endometrial hyperplasia restratified into EIN vs. nonEIN categories. 0/87 non-EIN and 43/110 EIN cases developed adenocarcinoma. 0.6 0.4 EIN (DS<1) 0.2 0.0 0 5 10 15 Followup Time, Months Figure 8: Long term cancer progression in women with EIN 20. Cancer outcomes that occur more than one year after EIN diagnosis are bonafide progression events from a premalignant to malignant phase of disease. Progression to cancer more than one year following EIN diagnosis is 45 times more likely compared to women without EIN. Note the tempo of cancer appearance indicates that it can take years for an EIN to evolve into adenocarcinoma.. 477 Women with “endometrial hyperplasia restratified into EIN vs. non-EIN categories. 2/359 non-EIN and 22/118 EIN cases developed adenocarcinoma. Cancer Free Survival 1.0 Benign (DS>1) 0.8 0.6 EIN (DS<1) 0.4 0.2 HR=45 0.0 0 \\ 50 100 150 Followup Time, Months 200 5.How Is EIN Diagnosed? (also see www.endometrium.org) EIN is diagnosed by a pathologist using routine (hematoxylin and eosin stained) sections prepared from a representative endometrial sample 27;28. It is extremely important to note that diagnostic accuracy may be severely compromised by exogenous progestin-containing hormonal therapies. For this reason, primary diagnosis or followup surveillance of a suspected EIN lesion should be based whenever possible on a sample obtained while the patient is not on therapeutic hormones. For those patients on progestins, diagnostic tissue can be obtained 2-4 weeks after stopping exogenous hormones, after completion of a withdrawal bleed. Although computerized morphometry has been a useful tool in identifying features characteristic of EIN, such equipment is not required for routine diagnosis. Rather, pathologist interpretation of stated criteria at a standard microscope is adequate. It should be noted that EIN is a precursor of endometrioid endometrial adenocarcinomas and is unrelated to the "Endometrial Intraepithelial Carcinoma" proposed 29 to be the earliest stages of papillary serous type endometrial adenocarcinomas. A framework for EIN Diagnosis is shown in Table I at the beginning of this sylabus. Notable is the clear separation of endometrial changes caused by unopposed estrogens, and carcinoma, from EIN. 1.Topography of EIN The distribution of a lesion is useful in distinguishing between the diffuse, field-wide effects, of an abnormal hormonal environment (anovulation, or persistent estrogen effect), surface changes secondary to stromal breakdown, and more focal EIN. Clonal origin from a single cell requires EIN lesions to begin as local processes within the endometrial compartment. Early EIN lesions are easily diagnosed by their contrast in architecture and cytology with the background from which they have emerged. Over time, EIN lesions may completely overrun the background endometrium, thereby removing the convenient lesion-to-background contrast in morphology which assist in EIN diagnosis. For this reason, or because of fragmentation, many EIN lesions must be diagnosed without the benefit of comparison with companion benign tissues. Exclusion of artifact and careful evaluation of the architectural and cytologic features of EIN usually permits accurate diagnosis in these instances. 2.EIN Diagnostic Criteria All of the diagnostic criteria of Table IV, listed as A-E below, must be met in order to make an EIN diagnosis. The entire slide should first be scrutinized under low magnification for localizing lesions, and if found, these areas examined under higher power to assess possible changes in cytology within the architecturally distinct focus. Widespread EIN lesions that have replaced the entire endometrial compartment tend to have a sufficiently atypical cytology that background normal endometrium is no longer required as a reference point for accurate diagnosis. Size, architecture, and cytology features are easy EIN diagnostic criteria. Much more difficult are exclusion of benign mimics and adenocarcinoma from the differential diagnosis. There are no simple rules for benign mimic exclusion. The broad universe of competing entities can only be recognized on sight by one who has the easy familiarity that comes with experience. Consistent demarcation of the EIN-adenocarcinoma threshold remains important clinically because it provides a basis for the clinician to evaluate the risks of electing hormonal rather than surgical therapy in younger patients who wish to retain fertility. Special diagnostic challenges, such as recognition of EIN within polyps, interpretation of subdiagnostically small or fragmented lesions, and interpretation of lesions with non-endometrioid differentiation have specific caveats presented below that should be carefully studied. Table IV: EIN Diagnostic Criteria. Modified after 5. EIN Criterion Architecture Cytology Size >1 mm Comments Area of Glands greater than Stroma Cytology differs between architecturally crowded focus and background, or clearly abnormal. Maximum linear dimension exceeds 1mm. Exclude mimics Benign conditions with overlapping criteria: Basalis, secretory, polyps, repair, etc.. Exclude Cancer Carcinoma if mazelike glands, solid areas, polygonal “mosaic-like” glands, myoinvasion, or significant cribriforming a.Architecture: Gland area exceeds stromal area: A cardinal architectural feature of endometrial precancers is glandular crowding, with a threshold quantitative cutoff for EIN lesions of less than half of the tissue area occupied by stroma (Volume Percentage Stroma). Areas with large dominant cysts should always be avoided in making this assessment. Although EIN is an epithelial disease, visual assessment of the glands themselves is complicated by frequent artifactual displacement from associated stroma, pale staining of most epithelia, and visual "shimmering" between gland epithelia and lumens. These may all be avoided by focusing on the stromal compartment which has the significant advantages of a more uniform composition throughout the specimen, and superior staining qualities. By focusing on the stroma itself only intact fragments in which stroma has not been avulsed from glands will be evaluated. Careful review of graphic and histologic examples of varying stromal densities will assist in training your eye to classify patient material as above or below the diagnostic threshold. EIN lesions tend to cluster with a median volume percentage stroma of about 40% and non-EIN (benign) lesions cluster at a median of approximately 75%. These differences are sufficiently great that visual assessment by a trained eye can be informative. b.Cytology of architecturally crowded area is different than background, or clearly abnormal: There is no absolute standard for cytologic features of EIN lesions, but the cytology of EIN is usually clearly demarcated as divergent from that of co-existing benign endometrial tissues in the same patient. The manner of cytologic change in EIN varies considerably from patient to patient, and can include but not be limited to, increased variation in nuclear size and contour, clumped or granular chromatin texture, change in nucleoli, change in nuclear/cytoplasmic ratio, and altered cytoplasmic differentiation. Stereotypical static descriptions of cytologic atypia, such as nuclear rounding and appearance of nucleoli are met in many but not all EIN lesions. In this sense, a fixed presentation of cytologic atypia is not a prerequisite for EIN. Attempts to define an absolute standard are confounded by the extreme morphologic plasticity of endometrial glandular cells under changing hormonal, repair, and differentiation conditions. Cytologic changes in some EIN lesions are manifest as a change in differentiation state to a tubal, mucinous, micropapillary, or eosinophilic phenotype. These must be distinguished from the scattered random pattern of hormonally, or surface located repair-induced “metaplasias.” Further details of how to interpret non-endometrioid EIN lesions are presented in the “Pitfalls” section below. In those cases with no normal glands for internal reference, it is necessary to assess the freestanding cytology of relevant fragments in the context of their architectural features. Some EIN lesions occupy the entire tissue sample, and should not be underdiagnosed for lack of a convenient benign gland in the area. c.Size >1mm in maximum dimension: Accurate EIN diagnosis requires a contiguous field of glands sufficiently large to enable reliable assessment of architecture. A minimum lesion size of 1 mm maximum dimension was required in the previous clinical outcome studies 19;20;22;24 for an EIN lesion to achieve elevated cancer risk. That area of an EIN lesion which meets architectural (gland area) and cytologic (changed) criteria for diagnosis must measure a minimum of 1mm in maximum dimension, a scale which usually encompasses more than 5-10 glands. Most biopsy formats produce tissue fragments in excess of 1.5-2mm. The size requirement must be met in a single tissue fragment, not added amongst multiple fragments. There is no formal evidence that once beyond the minimum 1mm, EIN lesions should be stratified by size, but if a lesion is discretely focal, it may be of interest to the clinician to know what fraction of the available curettings contain lesion. Individual or small clusters of cytologically altered glands have an undefined natural history and are best diagnosed descriptively (See Pitfalls section below). d.Exclusion of Benign Mimics Patients with one of the conditions listed below may still have an EIN, but this diagnosis should be made with careful consideration into how the coexisting factor(s) may modify the criteria for EIN diagnosis. If a specimen is refractory to confident diagnosis, a comment as to the nature of the problem may be useful in directing management. 1. Reactive changes caused by infection, physical disruption, recent pregnancy, or recent instrumentation. These can cause piling up of the epithelium, and loss of nuclear polarity.. 2. Artifactual gland displacement. Beware diagnosing an EIN lesion if the cytology is identical between areas with crowded compared to uncrowded glands! Many of these are artifactual disruptions where the stroma is sheared and glands pushed in apposition . 3. Persistent Estrogen Effect: Randomly scattered cysts of protracted estrogen exposure and occasional branching glands are commonly encountered in anovulatory or estrogen-exposed endometria. Gland density is uniformly irregular throughout the endometrial compartment, with occasional clusters of glands having a cytology identical to the uncrowded areas. These can be diagnosed as “Benign Endometrial Hyperplasia” if glands are significantly crowded, or in some mild cases as "disordered proliferative" endometrium. With increasing duration, microthrombi form and scattered stromal breakdown may be associated with epithelial piling along the collapsed stromal surfaces. 4. Mid to late secretory endometrium displays loss of nuclear polarity, nuclear enlargement, and variation in nuclear size which if measured objectively by computerized morphometry overlaps substantially with EIN lesions. Stromal responsiveness to progesterone is not homogenous at all endometrial depths. Lack of stromal pre-decidualization in the deeper functionalis and superficial basalis makes glands appear crowded, and these same glands may display a worrisome cytology and complicated saw-toothed luminal profiles 5. Endometrial polyps contain irregularly spaced glands in which scattered glands may differ from native endometrium due to their tendency to have reduced hormonal responsiveness. Benign polyps may also have low volume percentage stroma caused by cysts (senile polyps) or random aggregations of glands. Approximately 10% of EIN lesions, however, will present within an endometrial polyp and these must be diagnosed as described below in the “Pitfalls” section. 6. Endometrial breakdown is one of the most common settings for overdiagnosis of a benign endometrium as a precancer or cancer. Breakdown may follow an ovulatory or anovulatory cycle and persist into the transitional period between late menses and early proliferative endometrium. Altered cytology is due to piling up of epithelial cells unsupported by stroma, and associated nuclear changes such as loss of polarity which may be accentuated under certain fixation conditions which exaggerate chromatin texture (Bouin's fixative). e.Exclusion of Carcinoma Cancer may coexist with EIN in an individual patient, but should be always be separately diagnosed because current management of carcinoma differs from that for EIN. Keep in mind that absence of carcinoma in a tissue biopsy does not exclude the possibility of that the patient has a cancer which was unsampled during the biopsy procedure. An opinion should always be rendered based upon available material, and clearly stated. EIN is composed of individual glands lined by an epithelium one cell layer thick. The epithelium may be pseudostratified, but should not be cribriform or composed of solid areas of epithelial cells. Presence of any of the following features involving neoplastic glands is inconsistent with EIN, and a diagnosis of carcinoma should be entertained. 1. Meandering or “mazelike” lumens 2. Solid epithelium 3. Cribriform architecture. 4. “Mosaic” gland pattern of distorted polygonal glands with threadlike intervening stroma Myoinvasion. Unfortunately, myometrium is rarely available for evaluation in a biopsy or curettage specimen. Reference List 1. Mutter GL, Zaino RJ, Baak JPA, Bentley RC, Robboy SJ. The Benign Endometrial Hyperplasia Sequence and Endometrial Intraepithelial Neoplasia. Int J Gynecol Pathol 2007; 26:103-114. 2. Randall TC, Kurman RJ. Progestin treatment of atypical hyperplasia and welldifferentiated carcinoma of the endometrium in women under age 40. Obstet Gynecol Surv 1997; 90(3):434-440. 3. Mutter GL, Ince TA, Baak JPA, Kust G, Zhou X, Eng C. Molecular identification of latent precancers in histologically normal endometrium. Cancer Res 2001; 61:43114314. 4. Mutter GL, The Endometrial Collaborative Group. Endometrial intraepithelial neoplasia (EIN): Will it bring order to chaos? Gynecol Oncol 2000; 76:287-290. 5. Silverberg SG, Mutter GL, Kurman RJ, Kubik-Huch RA, Nogales F, Tavassoli FA. Tumors of the uterine corpus: epithelial tumors and related lesions. In: Tavassoli FA, Stratton MR, editors. WHO Classification of Tumors: Pathology and Genetics of Tumors of the Breast and Female Genital Organs. Lyon, France: IARC Press, 2003: 221-232. 6. Berman JJ, bores-Saavedra J, Bostwick D et al. Precancer: A conceptual working definition Results of a Consensus Conference. Cancer Detect Prev 2006; 30:387- 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 394. Esteller M, Garcia A, Martinez-Palones JM, Xercavins J, Reventos J. Detection of clonality and genetic alterations in endometrial pipelle biopsy and its surgical specimen counterpart. Lab Invest 1997; 76:109-116. Mutter GL, Chaponot M, Fletcher J. A PCR assay for non-random X chromosome inactivation identifies monoclonal endometrial cancers and precancers. Am J Pathol 1995; 146:501-508. Jovanovic AS, Boynton KA, Mutter GL. Uteri of women with endometrial carcinoma contain a histopathologic spectrum of monoclonal putative precancers, some with microsatellite instability. Cancer Res 1996; 56:1917-1921. Mutter GL, Boynton KA, Faquin WC, Ruiz RE, Jovanovic AS. Allelotype mapping of unstable microsatellites establishes direct lineage continuity between endometrial precancers and cancer. Cancer Res 1996; 56:4483-4486. Levine RL, Cargile CB, Blazes MS, Van Rees B, Kurman RJ, Ellenson LH. PTEN mutations and microsatellite instability in complex atypical hyperplasia, a precursor lesion to uterine endometrioid carcinoma. Cancer Res 1998; 58:3254-3258. Maxwell G, Risinger J, Gumbs C et al. Mutation of the PTEN tumor supressor gene in endometrial hyperplasias. Cancer Res 1998; 58:2500-2503. Mutter GL, Lin MC, Fitzgerald JT et al. Altered PTEN expression as a diagnostic marker for the earliest endometrial precancers. J Natl Cancer Inst 2000; 92:924-930. Duggan BD, Felix JC, Muderspach LI, Tsao J-L, Shibata DK. Early mutational activation of the c-Ki-ras oncogene in endometrial carcinoma. Cancer Res 1994; 54:1604-1607. Sasaki H, Nishii H, Takahashi H et al. Mutation of the Ki-ras protooncogene in human endometrial hyperplasia and carcinoma. Cancer Res 1993; 53:1906-1910. Mutter GL, Wada H, Faquin W, Enomoto T. K-ras mutations appear in the premalignant phase of both microsatellite stable and unstable endometrial carcinogenesis. Mol Pathol 1999; 52:257-262. Esteller M, Catasus L, Matias-Guiu X et al. hMLH1 Promoter Hypermethylation Is an Early Event in Human Endometrial Tumorigenesis. Am J Pathol 1999; 155(5):17671772. Mutter GL, Baak JPA, Crum CP, Richart RM, Ferenczy A, Faquin WC. Endometrial precancer diagnosis by histopathology, clonal analysis, and computerized morphometry. J Pathol 2000; 190:462-469. Dunton C, Baak J, Palazzo J, van Diest P, McHugh M, Widra E. Use of computerized morphometric analyses of endometrial hyperplasias in the prediction of coexistent cancer. Am J Obstet Gynecol 1996; 174:1518-1521. Baak JP, Mutter GL, Robboy S et al. The molecular genetics and morphometrybased endometrial intraepithelial neoplasia classification system predicts disease progression in endometrial hyperplasia more accurately than the 1994 World Health Organization classification system. Cancer 2005; 103(11):2304-2312. Hecht JL, Ince TA, Baak JP, Baker HE, Ogden MW, Mutter GL. Prediction of endometrial carcinoma by subjective endometrial intraepithelial neoplasia diagnosis. Mod Pathol 2005; 18:324-330. Baak JPA, Nauta J, Wisse-Brekelmans E, Bezemer P. Architectural and nuclear morphometrical features together are more important prognosticators in endometrial hyperplasias than nuclear morphometrical features alone. J Pathol 1988; 154:335341. Orbo A, Baak JP, Kleivan I et al. Computerised morphometrical analysis in endometrial hyperplasia for the prediction of cancer development. A long-term 24. 25. 26. 27. 28. 29. retrospective study from northern Norway. J Clin Pathol 2000; 53(9):697-703. Baak JP, Orbo A, van Diest PJ et al. Prospective multicenter evaluation of the morphometric D-score for prediction of the outcome of endometrial hyperplasias. Am J Surg Pathol 2001; 25(7):930-935. Mutter GL, Lin MC, Fitzgerald JT, Kum JB, Ziebold U, Eng C. Changes in endometrial PTEN expression throughout the human menstrual cycle. J Clin Endocrinol Metab 2000; 85:2334-2338. Stambolic V, Tsao MS, Macpherson D, Suzuki A, Chapman WB, Mak TW. High incidence of breast and endometrial neoplasia resembling human Cowden syndrome in pten+/- mice. Cancer Res 2000; 60(13):3605-3611. Mutter GL. Endometrial Intraepithelial Neoplasia: A new standard for precancer diagnosis. Cont Ob Gyn 2001; 46:92-98. Mutter GL. Histopathology of genetically defined endometrial precancers. Int J Gynecol Pathol 2000; 19:301-309. Ambros RA, Sherman ME, Zahn CM, Bitterman P, Kurman RJ. Endometrial intraepithelial carcinoma: A distinctive lesion specifically associated with tumors displaying serous differentiation. Hum Pathol 1995; 26:1260-1267. International Society of Gynecological Pathologists Society Companion Meeting United States and Canadian Academy of Pathology March 2, 2008 Endometrial Cancer Precursor Lesions: WHO is Better (?) Teri A. Longacre, MD Associate Professor of Pathology Stanford University Hospital & Medical Center Stanford University School of Medicine 1 Endometrial Cancer Precursor Lesions: Discussion Points Although there is little doubt that there are some forms of altered endometrial glandular proliferations that, when present, pose an increased risk of adenocarcinoma, the histologic definition of these risk lesions and their relative risk is the subject of ongoing controversy. Despite the advances in molecular biology of endometrial neoplasia, our knowledge of endometrial carcinoma and its precursor lesions is meager compared to precancer lesions in the breast and colon, and unlike these latter two organ systems, the acquisition of new knowledge is beset by formidable methodologic problems (3): 1) The morphologic definition of the target event (adenocarcinoma, usually grade 1 adenocarcinoma) is ill defined, subject to poor interobserver agreement, and a matter of ongoing debate. We use myoinvasion as the target event in our definition of grade 1 endometrial adenocarcinoma (i.e., only those proliferations that have been observed to be associated with myometrial invasion with significant frequency are considered “adenocarcinoma” with other lesser degrees of glandular proliferation considered “atypical” or “borderline”) (9). 2) Even if one were to use myoinvasive adenocarcinoma as the target event, the morphologic definition of myoinvasion (particularly superficial myoinvasion and adenocarcinoma involving adenomyosis) is subject to poor interobserver reproducibility. Ideally, the definition of an endometrial precursor lesion would be geared toward identifying those lesions that pose a significant risk for progression to clinically significant adenocarcinoma- i.e., myoinvasive carcinoma – not (to borrow a phrase used to designate very low risk lesions in the prostate) the pathologist’s carcinoma. 3) Unlike other organ systems, the putative precursor lesions in the endometrium are anatomically unstable; they can be shed spontaneously, or they can be reversed by a change in the patient's hormonal milieu, whether through alterations in physiology or alterations produced iatrogenically. 4) The endometrial sampling procedure is essentially a screening tool – not all of the endometrium may be represented in any given sampling, regardless of the technique and even when it is, the presence of a myometrial lesion cannot be assessed. Most pathologists and surgeons assume the presence of cancer in the myometrium (myoinvasive cancer) is associated with cancer in the endometrium. Fortunately, this is often the case, but there are occasional cancers that invade without an appreciable exophytic component (much like some invasive colorectal cancers in patients with longstanding ulcerative colitis). 5) Another unique feature of endometrial glandular proliferations is the variety of altered cytoplasmic differentiation or metaplastic patterns that commonly occur in both benign, precursor and cancer lesions. Since some of these altered differentiation patterns exhibit slightly more cytologic atypia than that which is 2 associated with standard endometrioid differentiation, definitions of precursor lesions must take into account this variable cytologic appearance. For example, ciliated change typically contains rounder nuclei, often with small nucleoli, and some of the problems with earlier definitions of atypical hyerplasia failed to take this into account. (A similar problem presents itself in the breast with apocrine lesions.) 6) Given the absence of evidence-based and consensus-driven diagnostic criteria, there is a perception that existing criteria for the diagnosis of endometrial cancer and its precursor lesions suffer from such poor reproducibility, that the willingness of the surgeon and patient to retain the organ involved by any putative precancerous lesion to wait out its natural history is low, particularly in the usual age group in which these lesions develop. 7) The use of surrogate markers (such as PTEN in the proposed endometrial intraepithelial neoplasia scheme) to inform our morphologic definitions of precancer and cancer must meet minimum criteria required for the use of a surrogate marker in other organ systems: i.e., be sensitive and specific, reproducible, and subject to independent confirmation. 8) Current terminology and definitions for endometrial precursor lesions are imprecise, but the introduction of new terminology should be conducted with caution. For example, the concept of “intraepithelial” is at best imprecise in the endometrium, given the absence of a bona fide basement membrane (such as is seen in the cervix), and at worst erroneous, given the emerging recognition in other organ systems of apparent shared properties of some in situ and invasive cancers. WHO Is Best? The WHO criteria for endometrial cancer precursor lesions are based on the study by Kurman and colleagues in 1985 (7). In that study, the risk of progression to carcinoma was 23% for atypical endometrial hyperplasia, whereas it was only 2% for non-atypical hyperplasia. Although the target lesion in this study was “carcinoma” and not “myoinvasive adenocarcinoma,” we know from previous studies that the definition(s) used for carcinoma in that study are, with minor exceptions, a reasonable stand-in for myoinvasive grade 1 endometrial cancer (9) The histologic criteria for complex atypical hyperplasia that were used in that study are well recognized and included a variety of cytologic and architectural features: nuclear enlargement, nuclear rounding, nucleoli, abnormal chromatin distribution (either dispersed or clumped), some degree of pleomorphism, loss of nuclear polarity, and a shift in the nuclear-to-cytoplasmic ratio in favor of the nuclei. The relative size of the nuclei was estimated by comparing them to the surrounding stromal cell nuclei or those of residual 3 normal epithelial elements. Mitotic figures were almost always present in atypical hyperplasia and often numerous, but abnormal division figures were sparse or absent. In the study published by Kurman and associates, once a patient had atypical hyperplasia, no further insight into risk was provided by grading the degree of atypia; that is, varying degrees of cytologic atypia were not reflected in a greater or lesser risk of adenocarcinoma once it was determined that the endometrium was architecturally complex and the glands were lined by cytologically atypical cells (6). This is not unexpected, given the narrow range of atypia that is present in these lesions. Subsequent reports using the Kurman criteria, provide additional evidence that approximately 20% to 30% of women with endometrial hyperplasia characterized by glands with marked architectural complexity and crowding, in addition to cytologic atypia, progress to a pattern that the investigators deemed morphologic adenocarcinoma (4). That is to say, not all precancers appear to progress to malignancy, either in the form of myoinvasion or clinical relapse, but a significant and diagnostic reproducible proportion do so despite the inherent difficulties in this diagnostically difficult range of endometrial glandular proliferations. Although the World Health Organization, which is largely based on data from the Kurman et al study, proposes a four-tiered classification, in most workers’ experience, the vast majority of cytologically atypical lesions are architecturally complex and therefore, the degree of cytologic atypia, despite its inherent reproducibility problems, may well be the best discriminator for precancer in this range of glandular proliferation. The degree of interobserver agreement for the diagnosis of “atypical hyperplasia” has been addressed by Kurman and coworkers (6). In that study, the only cytologic feature that was strongly associated with distinguishing hyperplasia (low risk for progression) from atypical hyperplasia (significant risk for progression) was the presence of nucleoli. The overall reproducibility for the diagnosis of “atypical hyperplasia” was moderate (kappa = 0.36 to 0.54) (6), while the overall reproducibility for the diagnoses of “hyperplasia” and “grade 1 carcinoma” were substantial. The interobserver reproducibility using the WHO criteria was more variable due to the use of 4 categories as opposed to 2, but was moderate overall and did not exceed that for the 2 category classification. Despite the utility of nucleoli in reproducibly distinguishing “hyperplasia” from “atypical hyperplasia” in that study, consensus diagnosis was achieved using a variety of pathways, not all of which entailed a conscious assessment for the presence of nucleoli (6). An experiential component combined with integration of multiple simultaneous evaluations appears to be inherent in the process of evaluating endometria for the presence of a risk lesion. Since the 2 tiered classification system of “hyperplasia” and “atypical hyperplasia” appears to perform as well as the 4 tiered WHO classification, the use of the 2 tiered system would appear to be preferable. Can We Do Better? 4 To appropriately answer the question as to what is the best method of diagnosing precancer in the endometrium, one has to pose the question: to what end? For the purposes of generating a molecular-based understanding of endometrial carcinogenesis, a classification scheme based on molecular correlates is of obvious scientific and possibly, epidemiologic value. For the purposes of clinical decision making, any taxonomic scheme that is used to classify precancer in the endometrium should reflect what is known about cancer risk – i.e., myoinvasive, clinically significant cancer risk. Ideally, the two end-points are complementary, but experience has shown that this is often not the case and standard histologic criteria based on outcome remain the gold standard. In absence of a carefully defined and consensus-driven target lesion to assess outcome and hence, inform our diagnostic criteria for endometrial cancer and precancer, studies such as those recently published from the GOG that reported a dismally and unacceptably low level of pathologist reproducibility for atypical hyperplasia and low grade carcinoma will continue to plague our literature and our profession (15-17). Given the formidable methodologic problems itemized above, one nihilistic view is that this is the best we can do. Another, perhaps more optimistic view is to search for molecular correlates of cancer and precancer to inform and improve on our standard histologic assessment. However, before we throw up our hands in defeat or rush to blindly embrace the molecules, it is important to remember that it may not always be necessary to render a finely tuned definitive diagnosis for this problematic set of lesions. In any given patient, it may be enough to give a best estimate of risk so that the treating physician and patient can make an informed decision to pursue a trial of hormonal therapy or proceed to a more definitive diagnostic (and therapeutic) procedure. In order to develop a more effective, real-world patient management program, further attempts to develop refined, evidence-based, and consensus-driven diagnostic criteria for these endometrial risk lesions should address these methodologic problems in well-designed, systematic, large-scale and independently confirmed studies. 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Cancer Lett 2001;167:39–48. 6. Kendall BS, Ronnett BM, Isacson C, et al. Reproducibility of the diagnosis of endometrial hyperplasia, atypical hyperplasia, and well-differentiated carcinoma. Am J Surg Pathol 1998;22:1012–1019. 7. Kurman R, Kaminski P, Norris H. The behavior of endometrial hyperplasia: a long-term study of "untreated" hyperplasia in 170 patients. Cancer 1985;56:403–412. 8. Kurman R, Norris H. Evaluation of criteria for distinguishing atypical endometrial hyperplasia from well-differentiated carcinoma. Cancer 1982;49:2547–2559. 9. Longacre TA, Chung MH, Jensen DN, et al. Proposed criteria for the diagnosis of well-differentiated endometrial carcinoma: a diagnostic test for myoinvasion. Am J Surg Pathol 1995;19:371–406 10. Montz FJ, Bristow RE, Bovicelli A, et al. Intrauterine progesterone treatment of early endometrial cancer. Am J Obstet Gynecol 2002;186:651–657. 11. Mutter GL. Endometrial intraepithelial neoplasia (EIN): will it bring order to chaos? The Endometrial Collaborative Group. Gynecol Oncol 2000;76:287–290. 12. Mutter GL, Baak JP, Crum CP, et al. Endometrial precancer diagnosis by histopathology, clonal analysis, and computerized morphometry. J Pathol 2000;190:462– 469. 13. Randall TC, Kurman RJ. Progestin treatment of atypical hyperplasia and well-differentiated carcinoma of the endometrium in women under age 40. Obstet Gynecol 1997;90:434–440. 14. Skov BG, Broholm H, Engel U, et al. Comparison of the reproducibility of the WHO classifications of 1975 and 1994 of endometrial hyperplasia. Int J Gynecol Pathol 1997;16:33–37. 15. Soslow RA. Problems with the current diagnostic approach to complex atypical endometrial hyperplasia. Cancer 2006;106:729-731. 16. Trimble CL, Kauderer J, Zaino R, et al. Concurrent endometrial carcinoma in women with a biopsy diagnosis of atypical endoemtrila hyperplasia: a Gynecologic Oncology Group Study. Cancer 2006;106:812-819. 17. Zaino RJ, Kauderer J, Trimble CL, et al. Reproducibility of the diagnosis of atypical endometrial hyperplasia: a Gynecologic Oncology Group Study. Cancer 2006;106:804811. 6 7 INTERNATIONAL SOCIETY OF GYNECOLOGICAL PATHOLOGISTS Sunday, March 2, 2008 – 1:30 p.m., Hyatt Regency Hotel, Denver, CO PATHOLOGY OF THE UTERINE CORPUS, PART 1: ENDOMETRIUM – CURRENT STATE OF THE ART ENDOMETRIAL CARCINOMA: CLASSIFICATION AND GENERAL FEATURES Jaime Prat, M.D., Ph.D., FRCPath. Hospital de la Santa Creu i Sant Pau Autonomous University of Barcelona, Spain For the last two decades, endometrial carcinoma has been subdivided into two major types (types I and II) based on epidemiology, conventional histopathology, and clinical behavior. Type I, which comprises approximately 80% of endometrial carcinomas newly diagnosed in the Western world, occurs predominantly in pre- and peri-menopausal women under unopposed estrogenic stimulation. These tumors are endometrioid carcinomas (EECs) that morphologically resemble normal endometrium and are frequently preceded by endometrial hyperplasia. They are usually confined to the uterus, exhibit low histological grade, and most patients are cured by hysterectomy. In contrast, type II endometrial carcinomas develop mainly in older post-menopausal women in whom the non-neoplastic endometrium is atrophic. These tumors are non-endometrioid carcinomas (NEECs), predominantly high grade serous or clear cell carcinomas, which are not associated with estrogen effect and are thought to derive from a malignant lesion designated ‘intraepithelial carcinoma’. Frequently, NEECs invade deeply into the myometrium and follow an aggressive clinical course. Also, it has been found that the genetic alterations carried by EECs differ from those of NEECs. Most were selected by analogy with colon cancer and were confirmed afterwards to occur in endometrial carcinoma. Recently, gene expression profiling has further expanded our knowledge of early genetic events and reinforced the clinicopathological subgroups originally defined by morphological and clinical features. However, even if a dualistic model may apply to typical cases, there is often overlap in the clinical, histopathological, immunohistochemical, and genetic characteristics of the tumors. Most endometrial carcinomas that are found in an atrophic endometrium are EECs, with a prognosis intermediate between the two types described above. Furthermore, it has been shown that some NEECs may develop from preexisting EECs as a result of tumor progression and, in such cases, the tumors may share histological and molecular features. Women with an inherited predisposition for endometrial neoplasia tend to develop the disease 10 years earlier than the general population and have a favorable prognosis. Most of these patients have hereditary non-polyposis colorectal carcinoma (HNPCC), an autosomal dominant disorder due to germline mutations in one of the DNA mismatch repair (MMR) genes. Although colorectal cancer predominates, endometrial carcinoma occurs in 30–60% of cases. References 1. Bockman JV. Two pathogenetic types of endometrial carcinoma. Gynecol Oncol 1983; 15: 10–7. 2. Amant F, Moerman P, Neven P, et al. Endometrial cancer. Lancet 2005; 366: 491–505. 3. American Cancer Society. Cancer Facts and Figures 2006. Atlanta: American Cancer Society, 2006. http://www.cancer.org/downloads/STT/CAFF2006PWSecured.pdf (accessed Nov 2006). 1 4. Jemal A, Siegel R, Ward E, et al. Cancer statistics, 2006. CA Cancer J Clin 2006; 56: 106–30. 5. Lax SF. Molecular genetic pathways in various types of endometrial carcinoma: from a phenotypical to a molecular-based classification. Virchows Arch 2004; 444: 213–23. 6. Catasu´ s Ll, Machin P, Matias-Guiu X, et al. Microsatellite instability in endometrial carcinomas clinicopathologic correlations in a series of 42 cases. Hum Pathol 1998; 29: 1160–4. 7. Lynch HT, de la Chapelle A. Genetic susceptibility to non-polyposis colorectal cancer. J Med Genet 1999; 36: 801–18. 8. Aarnio M, Sankila R, Pukkala E, et al. Cancer risk in mutations carriers of DNA-mismatch-repair genes. Int J Cancer 1999; 81: 214–8. 2 MADRID SEAP-07 - ADCA DE ENDOMETRIO: CLASIFICACIÓN 12/02/2008 ENDOMETRIAL CARCINOMA: CLASSIFICATION AND GENERAL FEATURES Jaime Prat, M.D., Ph.D., FRCPath. Hospital de la Santa Creu i Sant Pau Autonomous University of Barcelona, Spain Endometrial Carcinoma • Most common FGT cancer in western world • 4th most common cancer in women (6%) • Only 2% of cancer deaths in women • Postmenopause (75%) The two types of Endometrial Carcinoma Type I Type II Age Pre- and Perimenopausal Postmenopausal Unopposed Estrogen Present Absent Hyperplasia-Precursor Present Absent Grade Low High Myometrial Invasion Minimal Deep Histologic Type Endometrioid Nonendometrioid Behavior Stable Progressive Genetic alterations Microsat Instability P53 mutations, LOH Type I PTEN, Beta-catenin Modif from Bokhman JV. Gynecol Oncol 1983 Type II Endometrial Carcinoma Endometrioid ER p53 Non-Endometrioid Catasús Ll. et al. Hum Pathol 1998 1 MADRID SEAP-07 - ADCA DE ENDOMETRIO: CLASIFICACIÓN 12/02/2008 P53 Serous Ca Endometrioid Ca Endometrioid Ca Serous Ca BAX TGFβ-RII IGF-IIR MSH3 MSH6 MI, PTEN β-catenin Endometrioid Ca NE High grade endometrioid Ca Non-endometrioid Ca LOH P53 P53 Catasús Ll, et al. Hum Pathol 1998 2 INTERNATIONAL SOCIETY OF GYNECOLOGICAL PATHOLOGISTS Sunday, March 2, 2008 – 1:30 p.m. Hyatt Regency Hotel, Denver, CO PATHOLOGY OF THE UTERINE CORPUS, PART 1: ENDOMETRIUM – CURRENT STATE OF THE ART MOLECULAR BIOLOGY OF ENDOMETRIAL CARCINOMA Xavier Matias-Guiu MD, Department of Pathology and Molecular Genetics. Hospital Universitari Arnau de Vilanova University of Lleida, Spain. The molecular alterations involved in the development of endometrioid carcinomas (EEC) (type I) differ from those of non-endometrioid carcinomas (NEEC) (type II). Whereas EECs usually show microsatellite instability (MI), and mutations in the PTEN, k-RAS, PIK3CA, and beta-catenin genes, NEECs exhibit alterations of p53, loss of heterozygosity (LOH) on several chromosomes, as well as other molecular alterations (STK15, p16, Ecadherin and C-erb B2). Microsatellite instability (MI) was initially noted in cancers of patients with the hereditary non-polyposis colon cancer (HNPCC), but also in some sporadic colon cancers. EC is the second most common tumor found in HNPCC patients. MI has been demonstrated in 75% of EC associated with HNPCC, but also in 25-30% of sporadic EC. EC patients from HNPCC kindreds have an inherited germline mutation in either MLH-1, MSH-2, MSH-6 or PMS-2 (“first hit”); but EC develops only after the instauration of a deletion or mutation in the contralateral MLH-1,MSH-2, MSH-6 or PMS-2 allele (“second hit”) in endometrial cells. Once the two hits occur, the deficient mismatch repair role of the gene (MLH-1,MSH-2, MSH-6 or PMS-2) causes the acquisition of MI, and the development of the tumor. In sporadic EC, MI occurs more frequently in EEC (30%) than in NEEC. In sporadic tumors, MLH-1 inactivation by promoter hypermethylation is the main cause of mismatch repair deficiency. Abnormal methylation of MLH-1 may also be detected in atypical hyperplasias, suggesting that hypermethylation of MLH-1 may be an early event in the pathogenesis of EEC, preceding the development of MI. There are controversial data regarding the prognostic significance of MI, but there are some convincing evidence suggesting association with favourable outcome. The instauration of MI, the so-called mutator phenotype, in one cell has important molecular implications. The MI-associated mismatch repair deficiency leads to the accumulation of myriads of mutations in coding and noncoding DNA sequences. Short-tandem repeats, like microsatellites, are particularly susceptible to mismatch repair alterations, but they are predominantly located in non-coding DNA sequences; and the presence of subtle mutations (insertions or deletions) do not have consequences in the production of abnormal proteins. However, some small short-tandem repeats, like mononucleotide repeats, are sometimes located within the coding sequence of some important genes; (BAX, IGFIIR, hMSH3, and hMSH6 MBD4, CHK-1, Caspase-5, ATR, ATM, BML, RAD-50, BCL-10, Apaf-1) and they may be potential targets in the process of tumor progression of MI+, EC. Mutations in these tracts are interpreted as secondary events in the mutator phenotype pathway in cancers with MI, and usually alter the open reading frame of these genes, giving rise to abnormal or truncated proteins with altered functions. The tumor suppressor gene termed PTEN, located on chromosome 10q23.3, is frequently abnormal in endometrial carcinomas. LOH at chromosome 10q23 occurs in 40% of EC. Somatic PTEN mutations are also common in EC, and they are almost exclusively restricted to EEC, occurring in 37-61% of them. Interestingly, several groups have found a concordance between MI status and PTEN mutations; the mutations occur in 6086% of MI positive EEC, but in only 24-35% of the MI negative tumors. Such results have lead to the speculation that PTEN could be a likely candidate to be target for mutations in the MI positive EC. PTEN mutations have been detected in endometrial hyperplasias with and without atypia (19% and 21% respectively), both of them currently regarded as precursor lesions of EEC. Moreover, identical PTEN mutations have been detected in hyperplasias coexisting with MI positive EEC which suggests that PTEN mutations are early events in the development of EEC. There are controversial data regarding the prognostic significance of PTEN mutations in EC, but there are some results that suggest association with favourable prognostic factors. In agreement with Knudson’s two-hit proposal, LOH at 10q23 frequently coexists with somatic PTEN mutations. The coexistence of both alterations leads to activation of the PI3K/AKT pathway, which plays a key role in the regulation of cellular homeostasis. Activated AKT modulates the expression of several genes involved in suppression of apoptosis and cell cycle progression. However, it has been seen that these two do not necessarily coexist in the same tumors, challenging the two hit hypothesis for PTEN in EC. In a recent study of our group, we have detected any of these two genetic alterations in 57.7% of the tumors, but the coexistence of two genetic alterations (mutation and LOH or double mutations) was seen in only one third of the cases. In such study, the tumors that exhibited only one of these two alterations (mutation and LOH) were interpreted as having monoallelic inactivation of PTEN. Interestingly, the tumors with monoallelic inactivation of PTEN presented frequently mutations in the PIK3CA gene, which codes for the p110α catalytic subunit of PI3K. That means that the PI3K/AKT pathway can be altered by three different ways; 1) a tumor suppressor gene, such as PTEN, by showing somatic mutations and LOH, and 2) an oncogene, such as PI3KCA, by activating mutations, and 3) a single alteration in PTEN coexisting with an activating mutation in PI3KCA. Mutations in PIK3CA have been described in various tumors and may contribute to the alteration of the PI3K/AKT signalling pathway in endometrial carcinoma. PI3K is a heterodimeric enzyme consisting of a catalytic subunit (p110) and a regulatory subunit (p85).The PIK3CA gene, located on chromosome 3q26.32, codes for the p110α catalytic subunit of PI3K. A high frequency of mutations in the PIK3CA gene has been reported recently in several types of human cancer, including those of the colon, breast, ovary and stomach. The mutations are predominantly located in the helical (exon 9) and kinase (exon 20) domains. Oda et al described mutations in PIK3CA gene in endometrial carcinomas for the first time. In this series, PIK3CA mutations occurred in 36% of the cases, and coexisted frequently with PTEN mutations. Subsequent studies have shown that PIK3CA mutations are really frequent in EEC, in association with invasion, and adverse prognostic factors such as blood vessel invasion. These results suggest that PIK3CA mutations may contribute to activation of the PI3K/AKT pathway in endometrial carcinomas. The RAS-RAF-MEK-ERK signalling pathway plays an important role in tumorigenesis. Mutations in the RAS oncogene have been detected in many different types of tumors. The RAS superfamily of small GTP-binding proteins has a fundamental role in cell growth and differentiation, transcriptional regulation and apoptosis. The frequency of k-RAS mutations in EC ranges between 10 to 30%. In some series, K-RAS mutations have been reported to be more frequent in EEC showing microsatellite instability. In these tumors, K-RAS mutations are typically transitions, which may be preceded by abnormal DNA methylation. The fact that in EEC, microsatellite instability frequently coexists with K-RAS methylation-related transitions has lead to the suggestion that these two alterations are close related. During tumorigenesis, activated RAS is usually associated with enhanced proliferation, transformation and cell survival. BRAF, another member of the RAS-RAF-MEK-ERK pathways is very infrequently mutated in EC. RAS effectors like RASSF1A are supposed to have an inhibitory growth signal, which needs to be inactivated during tumorigenesis. Contradictory results between RASSF1A inactivation and K-RAS mutation have been obtained in different types of tumors. They were mutually exclusive events in colorectal and pancreatic cancer, but the correlation was not significant in lung cancer. Recent studies from our group have demonstrated that RASSF1A inactivation by promoter hypermethylation may contribute significantly to increased activity of the RAS-RAF-MEK-ERK signalling pathway. The beta-catenin gene (CTNNB1) maps to 3p21. Beta-catenin appears to be important in the functional activities of both APC and E-cadherin. Beta-catenin is a component of the E-cadherin-catenin unit, very important for cell differentiation, and maintenance of the normal tissue architecture. Beta-catenin is also important in signal transduction. Increased cytoplasmic and nuclear levels of beta-catenin produce transcriptional activation through the LEF/Tcf pathway. The APC protein down regulates beta-catenin levels by cooperating with the glycogen synthase kinase 3 beta (GSK-3beta), inducing phosphorylation of the serine-threonine residues coded in exon 3 of the beta catenin gene (CTNNB 1), and its degradation through the ubiquitin-proteasome pathway. Mutations in exon 3 of beta-catenin result in stabilization of the protein, cytoplasmic and nuclear accumulation, and participation in signal transduction and transcriptional activation through the formation of complexes with DNA binding proteins.Mutations in exon 3 of CTNNB1 with nuclear accumulation of beta-catenin occur in 14% to 44% of EC. They appear to be independent of the presence of MI, and the mutational status of PTEN and k-RAS. In all cases, the mutations were 2 homogeneously distributed in different areas of the tumors, which suggest that they do play a role in early steps of endometrial tumorigenesis. In fact, alterations in beta-catenin have been described in endometrial hyperplasias that contain squamous metaplasia (morules). Although, there was a good correlation between CTNNB 1 mutations and beta-catenin nuclear immunostaining, the presence of a cytoplasmic and nuclear beta-catenin immunoreactivity in some ECs that did not show a mutation in CTNNB suggests that alterations in other genes of the Wnt/beta-catenin/ LEF-1 pathway may be responsible for the stabilization and putative transcription activator role of beta-catenin in these tumors. There are controversial data regarding the prognostic significance of beta-catenin mutations in EC, but they probably occur is tumors with good prognosis. Apoptosis is a key process in the regulation of cellular homeostasis. Deregulation of apoptosis plays an important role in development and progression of cancer. The lack of response to such stimuli can originate a survival advantage, and the expansion of a population of neoplastic cells. Moreover, cells resistant to apoptosis are likely to escape the immune surveillance, but they may be also resistant to therapy. Apoptosis-resistant cells may expand while the patients receive anticancer treatment, and be responsible for relapse. Apoptosis can be initiated by two main mechanisms: the intrinsic pathway, which has its origin in the mitochondria, and the extrinsic apoptotic pathway, triggered by the activation of death receptors situated in the cell surface. A final common feature for execution of the apoptotic programme is the activation of a cascade of caspases, which are proteases that have a cysteine containing active site that cleaves protein substrates at specific amino acid motifs containing an aspartic acid residue. The ‘extrinsic pathway’, is activated by ligand-bound death receptors such as tumor necrosis factor (TNF), Fas or TRAIL receptors. After ligand binding, the activated death receptors recruit an adaptor protein named Fas Associated Death Domain (FADD). FADD consists of two protein interaction domains: a death domain (DD) and a death effector domain (DED). FADD binds to the receptor through interactions between DDs and to procaspase-8 through DED interactions to form a complex at the receptor called the Death Inducing Signalling Complex (DISC). Recruitment of caspase- 8 through FADD leads to its auto-cleavage and activation. Active caspase-8 in turn activates effector caspases such as caspase-3 causing the cell to undergo apoptosis by digesting upwards of a hundred or so proteins. One of the key regulators of this signalling is c-FLIP, which shares a high degree of homology with caspase- 8 but lacks protease activity. Thus it functions by competing with caspase-8 for binding to the DISC. In some type of cells this relatively simple pathway is enough to trigger apoptosis, but in other types of cells, the death receptor apoptotic pathway requires mitochondrial amplification. The BH3-only protein Bid is cleaved by caspase-8 and is then translocated to the mitochondria to activate the intrinsic pathway, thus connecting the two caspase activation pathways and amplifying the death receptor apoptotic signal. Thus, alterations in the mitochondrial pathway may affect the ability to induce apoptosis by death receptors. There are many evidences suggesting that alteration of apoptosis is important in development and progression of EC. Several of the molecular abnormalities that have been detected in EC may be associated with apoptosis deregulation. EEC show a high frequency of mutations in PTEN, which lead to constitutively active Akt, which in turn suppresses apoptosis triggered by various stimuli. Moreover, the recent evidence that NF-kB activation is frequent in endometrial carcinoma may explain the presence of apoptosis resistance by activation of target genes such as FLIP and Bcl-XL. p53 alterations, which are characteristic of NEEC, may also occur in endometrioid tumors, particularly in those neoplasms showing overlapping features between types I and II tumors; and they may have an impact in apoptosis at several different levels. Also, members of the Bcl-2 family of genes are abnormal in endometrial carcinoma. For example, BAX is a target gene for mutations in EEC with microsatellite instability, and may have a role in resistance to apoptosis in these tumors. Finally, several other proteins involved in apoptotic control (survivin) have also been shown to be abnormal in endometrial carcinoma. An important protein responsible for apoptosis resistance in endometrial carcinoma is FLIP. FLIP expression is frequent in endometrial carcinomas. A direct evidence of the role of FLIP in TRAIL apoptosis resistance on endometrial carcinoma cells is provided by treatment with specific siRNA targeting FLIP. Transfection of endometrial carcinoma cell lines with FLIP siRNA results in a marked decrease in cell viability after TRAIL exposition. This is accompanied by activation of both caspase-8 and caspase-3 suggesting activation of the extrinsic pathway. Recent studies have shown that FLIP may be regulated by a cellular complex composed by CK2-BRAF-KSR1. This is interesting because that will connect apoptosis resistance with the RAS-RAF-MEKERK signalling pathway. 3 In contrast to EEC, NEEC show p53 mutations (90%), inactivation of p16 (40%) and E-cadherin (80-90%), cerbB2 amplification (30%), alterations in genes involved in the regulation of the mitotic spindle checkpoint (STK-15) and loss of heterozygosity at multiple loci, reflecting the presence of chromosomal instability. While p53 mutations occur in 90% of NEEC, they are only present in 10-20% of EEC, which are mostly grade 3 tumors. The p53 protein can induce apoptosis or prevent a cell from dividing if there is DNA damage. Mutation of the p53 gene diminishes the cell’s ability to repair damage to DNA before entry to S-phase, leading to a greater chance that mutations will be fixed in the genome and passed to successive generations of cells. Inactivation of the cell cycle regulator p16 is also more frequent in NEEC (40%) than in EEC (10%). The underlying mechanism is not clear, but probably involves deletion and promoter hypermethylation. Reduced expression of E-cadherin is frequent in EC, and may be caused by LOH or promoter hypermethylation. In fact, LOH at 16q22.1 is seen in almost 60% of NEEC, but only in 22% of EEC. C-erbB2 overexpression and amplification are also seen more frequently in NEEC (43% and 29%) than in EEC. However, the most typical molecular feature of NEEC is chromosomal instability. This phenomenon is characterized by the presence of widespread chromosomal gains and losses, which reflect the presence of aneuploidy. cDNA arrays have demonstrated that NEEC usually show up-regulation of genes (STK-15, BUB1, CCNB2) that are involved in the regulation of the mitotic spindle checkpoint. One of them, STK-15, which is essential for chromosome segregation and centrosome functions, is frequently amplified in NEEC. None of the five main alterations of EEC (MI, and mutations in PTEN, k-RAS, PIK3CA and b-catenin) plays a significant role in NEEC. However, the occasional detection of these molecular alterations in NEEC, show that NEEC might develop by following two different pathways: 1) de novo, through p53 mutations, LOH at several loci, and some other, still unknown, gene alterations; or 2) dedifferentiation from pre-existing EEC. This hypothesis would explain the existence of mixed EEC-NEEC, and the presence of MI, as well as alterations in PTEN, K-RAS or beta-catenin in NEEC. According to this point of view, de novo NEEC (by far the most common situation) would fullfill the clinicopathologic and molecular features of NEEC (pure papillary serous or clear cell type morphology, old age, absence of estrogen stimulation, lack of prexisting endometrial hyperplasia, p53 mutations, lack of MI or PTEN mutations), whereas dedifferentiated NEEC, would exhibit overlapping features with EEC (mixed NEEC-EEC morphology, early age of presentation, evidence of estrogen stimulation or pre-existing hiperplasia, coexistence of p53 mutations and MI or PTEN mutations). cDNA array studies have demonstrated that the expression profiling of EEC is different from that of NEEC. In one study, 191 genes exhibited a >2-fold differences between 10 EECs and 16 NEECs. One of the genes, TFF3 was significantly up-regulated in EECs, while increased expression of FOLR was seen in NEECs. In a different study, a different expression profile was seen between EEC and NEEC, and the differences involved 66 genes. Interestingly, estrogen-regulated genes were up-regulated in EEC, whereas NEEC showed increased expression of genes involved in the regulation of the mitotic spindle checkpoint. A third study demonstrated differentially expression of 1,055 genes between EECs and serous carcinomas. Genes up-regulated in serous carcinomas were IGF2, PTGS1 and p16, while genes up-regulated in EEC included TFF3, FOXA2 and MSX2. A different study identified 315 genes that statistically differentiated EEC from NEEC. Moreover, a different expression profile was also found between EC associated with microsatellite instability and stable EC. 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Matias-Guiu, M.D. Department of Pathology and Molecular Genetics Hospital Arnau deVilanova University of Lleida, Spain K-ras 10-30% Beta-catenin 28-35% Microsatellite Instability Microsatellite Instability • Mismatch repair (MMR) genes (MLH1, MSH2) D5S107 D10S197 D12S79 D12S95 D18S58 N BAT-25 T N T N T N T N T • Frameshift mutations in target genes BAT-26 N • Hereditary Non-Polyposis Colorectal Cancer syndrome (Lynch II) T • Sporadic cancers: colon, stomach, pancreas, ovary, and endometrium T+N Endometrial Ca HNPCC (Lynch II) (n =118) Microsatellite Instability MSH2 MLH1 Endometrioid Ca Non-endometrioid Ca 37/109 (35%) 1/8 (12%) MSH2 MLH1 (Mutations)n T 1 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 Active Gene PRO MLH1 • Random Methylation Errors PRO Reduced Expression MLH1 Inactivation of DNA Mismatch Repair Genes by Promoter Hypermethylation in Endometrial Carcinoma MSP-hMLH1 E28 U M E21 U M E39 U E80 M U M E31 U E9 M U H2O IVD M U M U M • Clonal Selection of Cells • Further Random Methylation Errors PRO Silenced Gene MLH1 MI + 8/8 (100%) MI - 0/14 (0%) Esteller et al. 1999 Altered methylation MLH1 MI PTEN BAX TGF-BetaRII IGFIIR MSH3 MSH6 Caspase 5 H T D18S58 N T p16 U T T H H M U M APC E-Cadherin Mononucleotide repeat microsatellite Altered methylation MLH1 PTEN p16 APC E-Cadherin MI BAX TGF-BetaRII IGFIIR MSH3 MSH6 Caspase 5 Genes BAX MSH3 MSH6 TGF-BetaRII IGFIIR (G)8 (A)8 (C)8 (A)10 (G)8 2 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 Endometrial Ca, MI + (Frameshift mutations) BAX gene GGGGGGGG Caspase 5 45.4% BAX 41% Bcl-10 27% APAF-1 27% MSH3 25% IGFIIR 12.5% BLM 13.5% --------------------------Overall 77% Pr gene GGGGGGG Pr gene GGGGGGGGG Pr Catasús Ll. et al. Cancer 2000; 88:2290-7 BAX (G)8 T1 T1 T2 T2 BAX T1 T1 T2 N T T3 T4 Metastasis 3 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 Hyperplasia Carcinoma IGFIIR CASE 2487 N T PTEN CASE 2483 M N T CASE 2490 M T M Altered methylation MLH1 MI BAX RIZ IGFIIR MSH3 MSH6 Caspase 5 K-ras Beta-catenin MMP-7 Cyclin D1 Integrins Growth factor receptors PTEN alterations in the Endometrium Plasma Shc FAK Ptdins membrane (3,4,5)P3 PI 3-kinase Akt /PKB PTEN Adhesion and migration • Early event • PTEN-null glands (1%) in normal proliferative endometrium (43% of cases) • PTEN mutations in hyperplasia (15-55%) • PTEN mutations in carcinoma (30-60%) Survival proliferation and migration Endometrial Carcinoma PTEN mutations PTEN Mutations ET-28 T • Endometrioid • Non-Endometrioid T N 963-968 ins A 59/109 (54%) 0/5 (0%) ET-9 --------------------------------------------------------------- • MI+ ET-31 N 27/59 (46%) T N ET-47 ET-58 T T N N TTA GTA L146V P=0.005 4 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 Endometrial Hyperplasia PTEN mutations T H1 H2 H3 N N H T ET- 82 ET-140 5 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 MUTATION PTEN Endometrial Carcinoma IHC PTEN LOH N PROMOTER HYPERMETHYLATION T Alterations 42/78 54% Mutation LOH Methylation 34/78 24/78 6/34 44% 31% 18% One hit Two hits 23/78 19/78 29% 24% PTEN PTEN Endometrioid Carcinoma Biallelic inactivation Haplo-insufficiency Genetic Alterations PIK3CA 30% PTEN 30-60% Microsatellite Instability 20-30% K-ras 10-30% Beta-catenin 28-35% P ATP PIP2 Dephosphorylation PTEN PI3K Phosphorylation High frequency of coexistent mutations of PIK3CA and PTEN genes in Endometrial Carcinoma ADP PIP3 Oda K, et al Cancer Res, December 2005 AKT-P Cell Proliferation and Survival 1 5-Matias-Guiu ISGyP - Denver-08 PIK3CA mutations PIK3CA 36% (24/66) PTEN 56% (37/66) ---------------------------------------PIK3CA + PTEN 26% (17/66) 12/02/2008 PTEN alterations (Correlations in Endometrial Carcinoma) • PTEN mutations and MI • ↓ PTEN expression and ↑ phospho-AKT • ↓ PTEN expression and ↑ Survivin Oda K, et al Cancer Res, December 2005 Mutaciones PIK3CA 1 2 3 2 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 CTNNB1 mutation Endometrial Carcinoma CTNNB1 mutations • Endometrioid 15/59 (25.4%) • Non-Endometrioid 0/14 (0%) • MI+ • MI- 6/19 9/54 (31.5%) (16.6%) Beta-catenin nuclear accumulation Machin P et al, Hum Pathol 2002 MMP-7 expression Cyclin D1 expression Beta-Catenin Mutations in Endometrial Cancer CTNNB1 TCT TGT S37C EXON 3 Not associated with Microsat Instability More frequent in early stages? Good prognostic marker? 1 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 Genetic alterations in Atypical Endometrial Hyperplasia PTEN alt, K-RAS, and Beta-Catenin mutations, and MI n=78 AH (11) Number of alterations B-CAT PTEN RAS MSI 1 (9%) 4 (36%) 4 (36%) 1(9%) 0 0 0 AHWSM (14) 7 (50%) 0 27% 1 32% 2 27% (MI+PTEN: 13/19) 3 11% (MI+PTEN+RAS: 6/9) 4 3% Alterations Single alterations PTEN (n=42) (33%) 14 MI (n=30) (13%) 4 K-RAS (n=16) (12%) 2 Beta-catenin (n=14) (35%) 5 Bratchel, et al.; Am J Surg Pathol 2005 Growth Factors Death Factors NF-kappaB family members Death Receptors Adapter Protein Caspase 8 PI3K FLIP PTEN BAX (p65) AKT APAF-1 CYT-C Caspase 9 c- BCLxL NFKB Caspase 3 Survivin APOPTOSIS Adapted from Karin et al., 2004 Stimulus P p65 p65 IκB P Ub IκB p50 p50 p65 p65 p65 Ub p65 Ub P p50 IκB P p65 P p50 IκB P Proteosome p50 p52 1 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 NF-κB FLIP is frequenly overexpressed in Endometrial Adenocarcinoma (Endometrial Carcinoma) • • • • NF- κB expression is frequent P50-p65 co-expression (classic form) Bcl-3/p52 complexes are expressed Target genes are expressed (Cyclin D1, Flip, Bcl-xL) Pallares J et al: J Pathol 2004 204:569-577 CON siRNA FLIP Dolcet X et al: Lab Invest 2005 85:885-894 siRNA IK KLE Downregulation of FLIP by specific siRNA sensitizes to TRAIL-induced apoptosis 2 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 Growth Factors Death Factors NE Death Receptors Adapter Protein Caspase 8 EEC PI3K FLIP Supervised analysis between normal endometrium and Endometrioid Endometrial Carcinoma PTEN BAX AKT APAF-1 CYT-C Caspase 9 Hierarchical Clustering of 92 genes with differential expression patterns between NE and EEC (p<0.05), using a threshold of 2-fold BCLxL NFKB Caspase 3 Survivin APOPTOSIS CDH1 LOH in Endometrial Carcinoma E-Cadherin immunostaining in Endometrial Carcinomas N 16q22.1 D16S3057 D16S265 * D16S398 CDH1 T D16S496 D16S752 D16S496 Endometrioid Serous Preserved E-cadherin Reduced E-cadherin Focal loss of E-cadherin Reduced E-cadherin CDH1 LOH (22%) Endometrioid 41/82 (50.0%) Non-Endometrioid 27/31 (87.1%) 0.001 CDH1 LOH (57%) 1 5-Matias-Guiu ISGyP - Denver-08 12/02/2008 CYCLIN D1 and CYCLIN E NEEC Cyclin D1 CCND1 Cyclin E EEC G.Symbol A. Number Hierarchical Clustering of 55 genes with differential expression between EEC and NEEC (Supervised analysis, p<0.05) CCNE Cyclin D1 amplification FISH EEC 1/48 (2%) NEEC 5/19 (26%) Cyclin E amplification EEC 1/21 (5%) NEEC 5/12 (42%) Up-regulated genes in EEC Up-regulated genes in NEEC MGB2 10.4 CCNB2 2.6 LTF 6.5 DEK 2.6 NCOR1 3.9 STK15 2.5 END3 2.4 BUB1 2.5 p=0.033 Mean Fold Expression p=0.050 p<0.001 2 1.5 1 0.5 0 BUB1 EEC NEEC DEK STK15 25 22.5 20 17.5 15 12.5 10 7.5 5 2.5 0 NEEC EEC Breast cancer p<0.001 100 80 % Gene amplification 3 2.5 STK15 amplification in Breast and Gynecological cancer MGB NEEC 60 40 Ovary 20 Cervix Breast 0 TaqMan RT-PCR data validation Endometrial cancer Moreno-Bueno et al; Cancer Res 2003 Hyperplasia BAX TGFβ-RII IGF-IIR MSH3 MSH6 Carcinoma PTEN MI, PTEN β-catenin Altered methylation MLH1 MI PIK3CA BAX RIZ IGFIIR MSH3 MSH6 Caspase 5 Non-endometrioid Ca P53 P53 K-ras High grade endometrioid Ca Endometrioid Ca NE Beta-catenin Chromosome Instability LOH Amplification MMP-7 Cyclin D1 Cadherin E Cyclin D1 Cyclin E STK15 1 Objectives 2008 ISGP Symposium Morphology and prognostic factors in endometrial adenocarcinoma Richard J. Zaino, MD Hershey Medical Center Penn State University Hershey, PA [email protected] 1) Review the biology of the major types of endometrial adenocarcinoma 2) Examine the application of and significance of the CAP template for endometrial cancer 3) Examine the utility and limitations of the FIGO staging scheme for endometrial cancer 4) Examine prognostic factors in endometrial carcinoma MACROSCOPIC CAP approved (so it must be good for us) Surgical Pathology Cancer Case Summary (Checklist) Protocol revision date: January 2005 Based on AJCC/UICC TNM, 6th edition and FIGO 2001 Annual Report ENDOMETRIUM: Hysterectomy, With or Without Other Organs or Tissues Specimen Type ___ Hysterectomy ___ Radical hysterectomy (includes parametria) ___ Pelvic exenteration *Tumor Site *Specify location(s), if known: _____________________________ Tumor Size Greatest dimension: ___ cm *Additional dimensions: ___ x ___ cm ___ Cannot be determined (see Comment) Other Organs Present (check all that apply) ___ None ___ Right ovary ___ Left ovary ___ Right fallopian tube ___ Left fallopian tube ___ Urinary bladder ___ Vagina ___ Rectum ___ Other(s) (specify): _________________________ 1 Significance of maximum size of endometrial adenocarcinoma Relative few studies addressing prognostic significance Mariani et al, 2001 and 2002 size > 2cm is a predictor of lymphatic failure and distant failure by univariate analysis but not by multivariate analysis MICROSCOPIC Histologic Type ___ Endometrioid adenocarcinoma, not otherwise characterized ___ Endometrioid adenocarcinoma, secretory (variant) ___ Endometrioid adenocarcinoma, ciliated cell (variant) ___ Endometrioid adenocarcinoma, with squamous metaplasia ___ Adenosquamous carcinoma ___ Serous adenocarcinoma ___ Clear cell adenocarcinoma ___ Mucinous adenocarcinoma ___ Squamous cell carcinoma ___ Mixed carcinoma (specify types and percentages): ________________________ ___ Undifferentiated carcinoma Histologic Grade (if applicable) (Grading system below applies primarily to endometrioid carcinoma) ___ Not applicable ___ GX: Cannot be assessed ___ G1: 5% or less nonsquamous solid growth ___ G2: 6% to 50% nonsquamous solid growth ___ G3: More than 50% nonsquamous solid growth Pathologic classification of endometrial adenocarcinomas 1980 adenocarcinoma adenoacanthoma adenosquamous clear cell 2005 endometrioid endometrioid w squamous diff villoglandular secretory mucinous serous (UPSC) clear cell mixed 2 Endometrioid adenocarcinoma Smooth luminal border Estrogen receptor Secretory adenocarcinoma Villoglandular adenocarcinoma Adenocarcinoma with squamous differentiation 3 Mucinous adenocarcinoma Papillary serous carcinoma (UPSC) Clear cell adenocarcinoma Carcinoma with clear cells Carcinoma with clear cells 4 Two types of endometrial adenocarcinoma (Bokhman, 1981) Type I With hyperplasia (EIN) Hyperestrinism Endometrioid Well differentiated Steroid receptor + Good prognosis PTEN null/MSI uterine papillary serous carcinoma Type II No hyperplasia (EIC) No hyperestrinism Serous/CC Poorly differentiated Steroid receptor –/+ Poor prognosis p53 overexpression papillae Gaping glands Scalloped luminal border polyp Serous carcinoma Frayed or scalloped luminal border 5 MIB1 p53 ER Survival in endometrial adenocarcinoma (all stages) Tumor type 5 yr survival Endometrioid 80 – 90% UPSC 10 – 30% UPSC – patterns of spread Author Carcangiu Mallipeddi Lee Gitsch Carcangiu Cirisano Wheeler Goff Sherman Geisler sites of disease intrabdominal/small bowel nodes, bowel, omentum, cyto ovaries, nodes, peritoneum cyto, nodes, omentum, liver, dia adnexa, peritoneum, omentum, nodes nodes, ovaries, peritoneum, omentum ovary, omentum, bowel ovary, nodes, omentum, peritoneum nodes, cyto, ovary, omentum omentum, cyto, peritoneum, nodes 6 MICROSCOPIC Histologic Type ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ ___ Endometrioid adenocarcinoma, not otherwise characterized Endometrioid adenocarcinoma, secretory (variant) Endometrioid adenocarcinoma, ciliated cell (variant) Endometrioid adenocarcinoma, with squamous metaplasia Adenosquamous carcinoma Serous adenocarcinoma Clear cell adenocarcinoma Mucinous adenocarcinoma Squamous cell carcinoma Mixed carcinoma (specify types and percentages): ________________________ Undifferentiated carcinoma Histologic Grade (if applicable) (Grading system below applies primarily to endometrioid carcinoma) ___ Not applicable ___ GX: Cannot be assessed ___ G1: 5% or less nonsquamous solid growth ___ G2: 6% to 50% nonsquamous solid growth ___ G3: More than 50% nonsquamous solid growth Grade 1 Grade 1 Grade 2 7 Grade 2 Grade 3 Grade 3 (nuclei) Overall grade 2 Grade 3 Grade 1 (architecture) Grade 3 nuclei Overall grade 2 8 Stage I adenocarcinoma of the endometrium (FIGO 2003) Grade 1 2 3 5 year survival 92% 88% 75% Grading endometrial adenocarcinoma Two grades versus three FIGO – 3 grades, architecture +/- nuclear GOG – 3 grades, architecture Hachisuga -3 grades, nuclear (quantitative) Taylor et al – 2 grades, architecture (10% solid) Scholten – 2 grades, architecture (50% solid) Lax – 2 grades, architecture (solid, pattern, necrosis) Alkushi – 2 grades, architecture and nuclear Each prognosticates well Reproducibility of grading Alkushi (arch+nuclear) Nielsen (arch) Nielsen (nuclear) Zaino (arch) Zaino (nuclear) Taylor (arch) Lax (arch) Scholten (arch, using Lax) Inter-observer kappa 2 grade 3 grade 0.76 0.61 0.70 0.55 0.49 0.57 0.97 0.52 0.65 0.55 0.39 0.41 Lymphatic invasion lymphatic invasion 9 vascular pseudo-invasion Pathologic Staging (pTNM [FIGO]) Primary Tumor (pT) ___ pTX [--]: Primary tumor cannot be assessed ___ pT0 [--]: No evidence of primary tumor ___ pTis [0]: Carcinoma in situ pT1 [I]: Tumor confined to corpus uteri ___ pT1a [IA]: Tumor limited to endometrium ___ pT1b [IB]: Tumor invades less than one-half of the myometrium ___ pT1c [IC]: Tumor invades one-half or more of the myometrium pT2 [II]: Tumor invades cervix, but does not extend beyond uterus ___ pT2a [IIA]: Tumor limited to the glandular epithelium of the endocervix. There is no evidence of connective tissue stromal invasion. ___ pT2b [IIB]: Invasion of the stromal connective tissue of the cervix pT3 [III]: Local and/or regional spread as specified in T3a, T3b, N1, and FIGO IIIA, IIIB, and IIIC ___ pT3a [IIIA]: Tumor involves serosa, parametria, and/or adnexa (direct extension or metastasis) *___ pT3a [IIIA]: Tumor involves serosa and/or adnexa (direct extension or metastasis) and/or cancer cells in ascites or peritoneal washings ___ pT3b [IIIB]: Involvement of vagina (direct extension or metastasis), rectal or bladder wall (without mucosal involvement), or pelvic wall(s) (frozen pelvis) ___ pT4 [IVA]: Tumor invades bladder mucosa and/or bowel mucosa Regional Lymph Nodes (pN) ___ pNX: Cannot be assessed ___ pN0: No regional lymph node metastasis ___ pN1 [IIIC]: Regional lymph node metastasis Specify: Number examined: ___ Number involved: ___ Distant Metastasis (pM) ___ pMX: Cannot be assessed ___ pM1 [IVB]: Distant metastasis Stage I Corpus Cancer Pathologic Staging (pTNM [FIGO]) Primary Tumor (pT) ___ pTX [--]: Primary tumor cannot be assessed ___ pT0 [--]: No evidence of primary tumor ___ pTis [0]: Carcinoma in situ pT1 [I]: Tumor confined to corpus uteri ___ pT1a [IA]: Tumor limited to endometrium ___ pT1b [IB]: Tumor invades less than one-half of the myometrium ___ pT1c [IC]: Tumor invades one-half or more of the myometrium pT2 [II]: Tumor invades cervix, but does not extend beyond uterus ___ pT2a [IIA]: Tumor limited to the glandular epithelium of the endocervix. There is no evidence of connective tissue stromal invasion. ___ pT2b [IIB]: Invasion of the stromal connective tissue of the cervix pT3 [III]: Local and/or regional spread as specified in T3a, T3b, N1, and FIGO IIIA, IIIB, and IIIC ___ pT3a [IIIA]: Tumor involves serosa, parametria, and/or adnexa (direct extension or metastasis) *___ pT3a [IIIA]: Tumor involves serosa and/or adnexa (direct extension or metastasis) and/or cancer cells in ascites or peritoneal washings ___ pT3b [IIIB]: Involvement of vagina (direct extension or metastasis), rectal or bladder wall (without mucosal involvement), or pelvic wall(s) (frozen pelvis) ___ pT4 [IVA]: Tumor invades bladder mucosa and/or bowel mucosa Regional Lymph Nodes (pN) ___ pNX: Cannot be assessed ___ pN0: No regional lymph node metastasis ___ pN1 [IIIC]: Regional lymph node metastasis Specify: Number examined: ___ Number involved: ___ Superficial myometrial invasion 1) Is the distinction of non-invasive from inner half invasion reliable? 2) Should invasion be assessed in thirds or halves of myometrial thickness? 10 endometrium myometrium Superficial myometrial invasion Stage I Corpus Cancer significance of invasion Stage 5 year survival rates (FIGO, 2003) IA 92% inability to distinguish interIB 91% digitations from myo invasion IC 81% either outer third or outer half invasion highly significant (insufficient data to distinguish which is superior) Pathologic Staging (pTNM [FIGO]) Primary Tumor (pT) ___ pTX [--]: Primary tumor cannot be assessed ___ pT0 [--]: No evidence of primary tumor ___ pTis [0]: Carcinoma in situ pT1 [I]: Tumor confined to corpus uteri ___ pT1a [IA]: Tumor limited to endometrium ___ pT1b [IB]: Tumor invades less than one-half of the myometrium ___ pT1c [IC]: Tumor invades one-half or more of the myometrium pT2 [II]: Tumor invades cervix, but does not extend beyond uterus ___ pT2a [IIA]: Tumor limited to the glandular epithelium of the endocervix. There is no evidence of connective tissue stromal invasion. ___ pT2b [IIB]: Invasion of the stromal connective tissue of the cervix pT3 [III]: Local and/or regional spread as specified in T3a, T3b, N1, and FIGO IIIA, IIIB, and IIIC ___ pT3a [IIIA]: Tumor involves serosa, parametria, and/or adnexa (direct extension or metastasis) *___ pT3a [IIIA]: Tumor involves serosa and/or adnexa (direct extension or metastasis) and/or cancer cells in ascites or peritoneal washings ___ pT3b [IIIB]: Involvement of vagina (direct extension or metastasis), rectal or bladder wall (without mucosal involvement), or pelvic wall(s) (frozen pelvis) ___ pT4 [IVA]: Tumor invades bladder mucosa and/or bowel mucosa Regional Lymph Nodes (pN) ___ pNX: Cannot be assessed ___ pN0: No regional lymph node metastasis ___ pN1 [IIIC]: Regional lymph node metastasis Specify: Number examined: ___ Number involved: ___ Stage II Corpus Cancer Significance of true surgical pathologic staging: a GOG study (Creasman et al, 1999) 148/1180 with clinical stage II (+ECC) 66/148 with disease in the cervix 31/66 with extrauterine disease 35 (24%) with surgical stage II Recurrence rates at 5 years: IIA – 18% IIB – 21% 11 Stage II Corpus Cancer 5 year survival - 75%, lower than Stage I (FIGO results, 2003) More often associated with higher grade, deep myometrial invasion, and lymphatic invasion than Stage I Insufficient data to determine whether Stage II is a significant prognosticator by multivariate analysis (probably not) Stage II Corpus Cancer IIA IIB endocervical gland involvement cervical stromal invasion Definitions applied in various publications: IIA - surface epithelium only (Jordan) IIA – gland involvement only (Fanning, Eltabbakh, Prat) IIA – confined to endocervical epithelium (mucosa) (Clement and Young) -but endocervix lacks a mucosa diagnostic reproducibility probably low (never tested) how does it involve glands only? Pathologic Staging (pTNM [FIGO]) Primary Tumor (pT) ___ pTX [--]: Primary tumor cannot be assessed ___ pT0 [--]: No evidence of primary tumor ___ pTis [0]: Carcinoma in situ pT1 [I]: Tumor confined to corpus uteri ___ pT1a [IA]: Tumor limited to endometrium ___ pT1b [IB]: Tumor invades less than one-half of the myometrium ___ pT1c [IC]: Tumor invades one-half or more of the myometrium pT2 [II]: Tumor invades cervix, but does not extend beyond uterus ___ pT2a [IIA]: Tumor limited to the glandular epithelium of the endocervix. There is no evidence of connective tissue stromal invasion. ___ pT2b [IIB]: Invasion of the stromal connective tissue of the cervix pT3 [III]: Local and/or regional spread as specified in T3a, T3b, N1, and FIGO IIIA, IIIB, and IIIC ___ pT3a [IIIA]: Tumor involves serosa, parametria, and/or adnexa (direct extension or metastasis) *___ pT3a [IIIA]: Tumor involves serosa and/or adnexa (direct extension or metastasis) and/or cancer cells in ascites or peritoneal washings ___ pT3b [IIIB]: Involvement of vagina (direct extension or metastasis), rectal or bladder wall (without mucosal involvement), or pelvic wall(s) (frozen pelvis) ___ pT4 [IVA]: Tumor invades bladder mucosa and/or bowel mucosa Regional Lymph Nodes (pN) ___ pNX: Cannot be assessed ___ pN0: No regional lymph node metastasis ___ pN1 [IIIC]: Regional lymph node metastasis 12 Stage III Corpus Cancer: Are all forms of Stage IIIA disease equivalent? Positive peritoneal cytology Direct invasion to uterine serosa Adnexal spread or metastases Stage IIIA Corpus Cancer + peritoneal cytology Milosevic et al, 1992, pooling of literature 17 studies, 3800 patients; incidence – 11% Often associated with adnexal or nodal spread, high grade, deep invasion Univariate analysis – increased risk of recurrence in most studies Multivariate analysis (5 studies) – decreased survival in 3/5 + cyto as an isolated poor prognostic factor is rare Stage IIIA Corpus Cancer + adnexal involvement (Connell et al, 1999) 5 year DFS – 37% overall Often associated with higher grade, lymphatic invasion, other extrauterine disease 5 year DFS - 71% without other extrauterine spread Stage IIIA Corpus Cancer + serosal involvement (SI) (Ashman et al, 2001) 5 year DFS – 29% 5 year DFS – 20%, SI + other extrauterine sites 5 year DFS – 42%, SI only Are all forms of Stage IIIA disease equivalent? 5 year recurrence free survival (Mariani et al, 2002) + cytology only 79% + adnexa/uterine serosa 57% (p = 0.04) (Tebeu et al, 2004) + cytology only + adnexa/uterine serosa 91% 50% 13 Stage III Corpus Cancer Stage IIIB – vaginal metastases Very rare, (less than 1% of corpus cancer and about 2% of stage III pts) Vaginal mets often associated with nodal or distant metastases Prognosis poor – 5 year survival about 25% Pathologic Staging (pTNM [FIGO]) Primary Tumor (pT) ___ pTX [--]: Primary tumor cannot be assessed ___ pT0 [--]: No evidence of primary tumor ___ pTis [0]: Carcinoma in situ pT1 [I]: Tumor confined to corpus uteri ___ pT1a [IA]: Tumor limited to endometrium ___ pT1b [IB]: Tumor invades less than one-half of the myometrium ___ pT1c [IC]: Tumor invades one-half or more of the myometrium pT2 [II]: Tumor invades cervix, but does not extend beyond uterus ___ pT2a [IIA]: Tumor limited to the glandular epithelium of the endocervix. There is no evidence of connective tissue stromal invasion. ___ pT2b [IIB]: Invasion of the stromal connective tissue of the cervix pT3 [III]: Local and/or regional spread as specified in T3a, T3b, N1, and FIGO IIIA, IIIB, and IIIC ___ pT3a [IIIA]: Tumor involves serosa, parametria, and/or adnexa (direct extension or metastasis) *___ pT3a [IIIA]: Tumor involves serosa and/or adnexa (direct extension or metastasis) and/or cancer cells in ascites or peritoneal washings ___ pT3b [IIIB]: Involvement of vagina (direct extension or metastasis), rectal or bladder wall (without mucosal involvement), or pelvic wall(s) (frozen pelvis) ___ pT4 [IVA]: Tumor invades bladder mucosa and/or bowel mucosa Regional Lymph Nodes (pN) ___ pNX: Cannot be assessed ___ pN0: No regional lymph node metastasis ___ pN1 [IIIC]: Regional lymph node metastasis Specify: Number examined: ___ Number involved: ___ Stage III Corpus Cancer Stage III C – pelvic/paraaortic nodal mets (Mariani et al, 2002) Stage IIIC often are also Stage IIIA/IIIB 5 year DFS – 33% Stage IIIC with IIIA/B mostly extranodal failures 5 year DFS – 68% Stage IIIC without IIIA/B mostly nodal failures Stage III Corpus Cancer Stage III C – pelvic/paraaortic nodal mets 5 year DFS – about 65-80% + pelvic node 5 year DFS – about 30% + paraaortic node Significant survival differences between microscopic and grossly positive nodes, resected vs non-resected disease, radiated vs non-irradiated nodal beds, and capsular invasion and desmoplasia Tentative staging conclusions Stage IA cannot reliably be distinguished from Stage IB pathologically in many cases Stage IIA and IIB are poorly defined pathologically and may not differ prognostically Stage II is probably not prognostically significant Stage III disease is very heterogeneous Stage IIIA alone is heterogeneous + cytology alone is rare and probably significant but with small effect (85%) + adnexa is more significant (70%) + uterine serosa carries a poor prognosis (30%) 14 Surgical Staging of Corpus Cancer (EGO, 2008) Tentative staging conclusions Stage IIIB (vaginal mets) very rare, poor prognosis (25%) Stage IIIC + pelvic nodes significant (70%) + paraaortic nodes significantly worse (30%) grossly positive nodes; capsular invasion and desmoplasia; other extrauterine sites associated with a much worse outcome (Stage IIIC limited to nodes usually fails in nodal area) Additional prognostic factors Flow cytometry Steroid receptors Markers of apoptosis (BCL-2) Markers of proliferation Tumor suppressor genes and oncogenes Markers of angiogenesis Potential utility Steroid receptors BCL-2 MIB1 Her2/neu Microvessel density Stage IA G123 IB G123 IIA IIB IIIA1 IIIA2 IIIB1 IIIB2 IVA IVB Characteristics invasion to endometrium/inner half of myometrium invasion to outer half of myometrium positive peritoneal cytology uterine serosa, adnexal or other pelvic spread pelvic nodal metastasis (micro) pelvic nodal metastasis (gross) paraaortic nodal metastasis (micro) paraortic nodal metastasis (gross) vaginal metastases, invasion of bladder or bowel mucosa distant, intraabdominal or inguinal node metastases Demonstrated utility FIGO stage Histologic grade Cell type Depth of myometrial invasion Lymphatic invasion Tumor ploidy p53 Unlikely utility PCNA S-phase fraction AgNOR count 15 Objectives 1) Review the biology of the two major types of endometrial adenocarcinoma 2) Examine the application of and significance of the CAP template for endometrial cancer 3) Examine the utility and limitations of the FIGO staging scheme for endometrial cancer 4) Examine prognostic factors in endometrial carcinoma 16 Endometrial Cancer: Who Needs Lymphadenectomy A Mariani, SC Dowdy, MB Jones, WA Cliby, BS Gostout, TO Wilson, KC Podratz An exigent need exists for a paradigm shift in the management of endometrial cancer. The continuing debate as to whether to perform lymphadenectomy (LND) versus radiotherapy (RT) exemplifies a modality-based approach to treatment rather than using a disease-based care pathway. The primary objectives for a revised paradigm should be to optimize outcomes through minimizing both overtreatment and undertreatment. Overtreatment can be minimized by identifying patients not requiring LND or RT and undertreatment minimized by identifying patients benefiting from one or both modalities. Disease-based therapy should be predicated on anticipated patterns of failure predicted by pathologic (and/or molecular) determinants. The merits of LND are diagnostic, prognostic, therapeutic and predictive of the requirement for adjuvant therapy. LND should be reserved for patients at risk for node metastasis. An absence of nodal involvement and a 5 year survival of 100% were observed in a cohort of 123 consecutive patients fulfilling the following criteria: Grade 1/2, endometrioid, < 50% myometrial invasion and primary tumor diameter < 2 cm. Accordingly, LND is not indicated in these low risk patients. This group accounted for 20% of the overall population and 29% of endometrioid patients. Node positivity in the remaining endometrioid population (71%) was 17%. Thus, for all other endometrioid as well as nonendometrioid patients we recommend a systematic LND up to the renal vessels. The independent risk factors dictating pelvic sidewall failure include cervical stromal invasion and lymph node metastasis (LNM); 5 year failure rates are <1 % in the absence of these factors and 26% when either or both were present (p<0.001) despite traditional modality based treatment. Pelvic sidewall failures at 5 years in patients with positive nodes approximated 10% in our cohort treated with combined systematic LND and adjuvant RT compared to >50% for patients treated with either LND or RT alone (p<0.01). Furthermore, in the presence of pelvic LNM, the greater majority of patients had either paraaortic LNM at the time of surgery or subsequently failed in the paraaortic area. Importantly, in a recently completed prospective assessment, the majority of patients with paraaortic node involvement had negative nodes below the inferior mesenteric artery (IMA) while the greater majority had documented positive nodes above the IMA. Hence, LND is a determinant for adjuvant therapy including the requirement for extended RT fields including the paraaortic area whenever LNM is detected in either the pelvic or paraaortic node bearing regions. REFERENCES 1. Podratz KC, Mariani A, Webb MJ. Staging and therapeutic value of lymphadenectomy in endometrial cancer -Editorial. Gynecol Oncol 70:163-164, 1998. 2. 3. 4. 5. 6. 7. 8. 9. 10. Mariani A, Webb MJ, Galli L, Podratz KC. Potential therapeutic role of para-aortic lymphadenectomy in node-positive endometrial cancer. Gynecol Oncol 76:348-356, 2000. Mariani A, Sebo TJ, Katzmann JA, Keeney GL, Roche PC, Lesnick TG, Podratz KC. Pretreatment assessment of prognostic indicators in endometrial cancer. Am J Obstet Gynecol 182:1535-44, 2000. Mariani A, Webb MJ, Keeney GL, Haddock MG, Calori G, Podratz KC. Low-risk corpus cancer: Is lymphadenectomy or radiotherapy necessary? Am J Obstet Gynecol 182:1506-19, 2000. Mariani A, Webb MJ, Rao S, Lesnick T, Podratz KC. Significance of pathologic patterns of pelvic lymph node metastases in endometrial cancer. Gynecol Oncol 80:113-120, 2001. Mariani A, Webb MJ, Keeney GL, Podratz KC. Routes of lymphatic spread: A study of 112 consecutive patients with endometrial cancer. Gynecol Oncol. 81:100104, 2001. Mariani A, Webb MJ, Keeney GL, Aletti G, Podratz KC. Predictors of lymphatic failure in endometrial cancer. Gynecol Oncol.84:437-442, 2002. Mariani A, Keeney GL, Aletti G, Webb MJ, Haddock MG, Podratz KC. Endometrial carcinoma: paraaortic dissemination. Gynecol Oncol 92:833-8, 2004. Mariani A, Sebo TJ, Katzmann JA, Roche PC, Keeney GL, Lesnick TG, Podratz KC. Endometrial cancer: can nodal status be predicted with curettage. Gynecol Oncol 96:594-600, 2005. Mariani A, Dowdy SC, Cliby WA, Haddock MG, Keeney GL, Lesnick TG, Podratz KC. Efficacy of systematic lymphadenectomy and adjuvant radiotherapy in nodepositive endometrial cancer patients. Gynecol Oncol 101:200-208, 2006. Endometrial Cancer Surgical Staging Endometrial Cancer Surgical Staging Role of Lymphadenectomy • Definitive Staging • TAH/BSO/Peritoneal cytology • Pelvic/Paraaortic LND* • Biopsy/Omentectomy • Cytoreduction (Rx) Karl Podratz MD PhD FACS *LND = Lymph node dissection Endometrial Cancer Endometrial Cancer Role of Lymphadenectomy vs Radiotherapy Annual Incidence Cases and Deaths Year ACS Estimates* Cases Deaths 1987 2007 35,000 2,900 39,080** 7,400*** • Modality-based therapy* • Lymphadenectomy • Radiotherapy *Traditions, physician preferences, suboptimal study designs, etc. Endometrial Cancer Role of Radiotherapy and Lymphadenectomy • Treatment paradigm shift • Minimize overtreatment –Identify pts not requiring LND and/or RT • Minimize undertreatment –Identify pts benefiting from LND and/or RT • Maximize outcomes *Ca 1987; CA 2007 **11.7% increase; ***155% increase Endometrioid Endometrial Cancer Role of Radiotherapy and Lymphadenectomy • Modality-based therapy • Radiotherapy vs. lymphadenectomy • Uterine histology • Disease-based therapy • Based on patterns of failure » Predicted by pathologic determinants • Selective Lymphadenectomy • Selective Radiotherapy • Selective Chemotherapy 1 Endometrial Cancer Endometrial Cancer Selective Lymphadenectomy Selective Lymphadenectomy (not sampling) • Lymphadenectomy not indicated* • Lymph Node Dissection (LND) • Low risk: Not indicated • All others: Systematic • Low risk: »Endometrioid »G 1&2 »MI < 50% »PTD < 2 cm *Mariani et al. Am J Ob Gyn 2000 Endometrioid Endometrial Cancer Endometrioid Endometrial Cancer Low risk: G1/2, < 2 cm, < 50% MI Low Risk: G 1/2, MI < 50%, PTD < 2 cm Treatment^ Hysterectomy only Hyst + LND* +/or RT** Total Pt (no.) 59 64 123 % 5 yr Survival 100 100 ^3/113 recurred (vagina) without RT; all salvaged • Lymphadenectomy not indicated • 20% Over all population* • 29% Endometrioid patients* *Mariani et al. Am J Ob Gyn 2000 *All nodes negative; **10 RT; 7 for PPC Mariani et al. Am J Ob Gyn 2000 Endometrioid Endometrial Cancer Selective Lymphadenectomy • Lymphadenectomy not indicated • Low risk: G 1/2, MI < 50%, PTD < 2 cm • Systematic Lymphadenectomy • All others (not low risk) • 17% positive nodes 2 Endometrial Cancer Failures Endometrial Cancer Failures Pelvic Lymphatic Failures Lymphatic Failures Lymphatic failures according to risk factors Lymphatic failures according to risk factors Lymphatic Failure rate P Site % at 5 years Value Pelvic Sidewall Low risk High risk* <1 26 <0.001 Low risk = absence of high risk factors High risk = *CSI and/or LN mets Lymphatic Site(s) Pelvic Sidewall Low risk High risk* Failure rate % at 5 years P Value <1 26 <0.001 1 33 <0.001 Para-aortic area Low risk High risk** Low risk = absence of high risk factors High risk = *CSI and/or LN mets; **LN mets only Endometrial Cancer Failures Endometrial Cancer Failures Paraaortic Lymphatic Involvement Paraaortic Lymphatic Involvement • 33% para-aortic failures with pelvic and/or para-aortic LN mets • 47% with positive pelvic LN either had para-aortic LN mets or para-aortic failures * • 47% para-aortic LN mets or para-aortic failures with pelvic LN mets* *Mariani et al 2002 (Mayo series) *Mariani et al 2002 (Mayo series) Endometrial Cancer Endometrial Cancer Surgical Management* Quality Assessment LND Number paraaortic nodes removed per surgeon during phase I • Hysterectomy, BSO, Peritoneal Cytology, Pelvic/Para-aortic lymphadenectomy (up to 35 30 • Omit lymphadenectomy if Grade 1 or 2, endometrioid, MI < 50%, and PTD < 2 cm • Omit lymphadenectomy if non-invasive endometrioid regardless of PTD or grade • Separately submit nodes above & below IMA • If non-endometrioid, add complete omentectomy, appendectomy, peritoneal biopsies, cytoreduction *Mayo prospective accrual 1/2004 to 12/2006 numberpa renal vessels) 25 20 15 10 5 1 2 3 surgeon 4 5 6 7 Mayo QI Project 3 Endometrial Cancer Endometrial Cancer Quality Assessment LND Surgical Management* Number paraaortic nodes removed per surgeon during phase II 35 • Objectives of Prospective Rx Algorithm 30 • Prevalence Pelvic LN mets according to histologic subtype • Prevalence Para-aortic LN mets with lymphatic dissemination • Para-aortic metastatic site frequency as function of IMA numberpa 25 20 15 10 5 0 1 2 3 surgeon 4 5 6 7 Mayo QI Project *Mayo prospective accrual 1/2004 to 12/2006 Endometrial Cancer Endometrial Cancer Surgical Management* Surgical Management* • 422 patients • 112 (27%) LND not indicated • 281 at-risk patients LND • 15 pelvic (P) LND only • 1 para-aortic (PA) LND only • Median # nodes harvested »90 (80%) no LND »22 (20%) had LND (all neg) • 310 (73%) required LND » Pelvis 35 » Para-aorta 17 »29 (9%) no LND »281 (91%) had LND • 63 (22%) positive nodes *Mayo prospective accrual 1/2004 to 12/2006 Node Site PA P+PA P *Mayo prospective accrual 1/2004 to 12/2006 Endometrial Cancer Endometrial Cancer Surgical Staging Surgical Staging # Pos n=63* 10 29 24 % 16 46 38 PA 62% P 84% *63/281 (22%) at-risk patients had positive nodes Mayo prospective accrual 1/2004 to 12/2006 Node Site PA P+PA P # Pos n=57* 9 29 19 % 16 51 33 PA 67% P 84% *57/265 (22%) at-risk pts had both P+PA LND & + nodes Mayo prospective accrual 1/2004 to 12/2006 4 Endometrial Cancer Surgical Staging* Paraaortic Node Metastases Skipping Common Iliac Nodes • 63 (22%) at-risk pts Pos Nodes • 84% + pelvic nodes • 67% + paraaortic nodes 71% »71% com Iliacs neg *Mayo prospective accrual 1/2004 to 12/2006 Endometrial Cancer Endometrial Cancer Surgical Staging* Surgical Staging* • 63 (22%) at-risk pts Pos Nodes • 84% + pelvic nodes • 67% + paraaortic nodes »71% com Iliacs neg »60% neg below IMA • 63 (22%) at-risk pts Pos Nodes • 84% + pelvic nodes • 67% + paraaortic nodes »71% com Iliacs neg »60% neg below IMA »77% + above IMA *Mayo prospective accrual 1/2004 to 12/2006 Paraaortic Node Metastases Metastasis above IMA *Mayo prospective accrual 1/2004 to 12/2006 INVASION OF THE GONADAL VESSELS or surrounding soft tissue 77% IMA 28% 5 Endometrial Cancer Surgical Staging* Non-Endometrioid Endo Ca (NEEC) Role of Surgical Staging • Surgical Staging required • Lymphadenectomy up to IMA only • 38-46% PA node Positive cases missed • Managed as ovarian • 422 EC pts Rx’ed surgically* • 82 (19%) NEEC • 62% cases node positive below IMA are node positive above IMA *Mayo prospective accrual 1/2004 to 12/2006 – 37% Macro extra-uterine disease – 21% Micro extra-uterine disease » 25% noninvasive – 40% Node metastasis *Mayo prospective series: 1/04-12/06 Endometrioid Endometrial Cancer Endometrioid Endometrial Cancer Surgical Management* Surgical Management* • 340 (81%) patients • 112 (33%) LND not indicated »90 (80%) no LND »22 (20%) had LND (all neg) • 228 (67%) required LND »19 (8%) no LND »209 (92%) had LND • 209 at-risk patients LND • 11 pelvic LND only • Median # nodes harvested » Pelvis 36 » Para-aorta 17 • 34 (16%) positive nodes »10% population *Mayo prospective accrual 1/2004 to 12/2006 *Mayo prospective accrual 1/2004 to 12/2006 Endometrioid Endometrial Cancer Endometrioid Endometrial Cancer Surgical Staging Surgical Staging* Node Site # Pos n=32* % PA P+PA P 6 14 12 19 44 37 PA 63% P 81% *32/198 (16%) at-risk pts had both P+PA LND & + nodes Mayo prospective accrual 1/2004 to 12/2006 • 32 (16%) at-risk pts Pos Nodes • 81% + pelvic nodes • 63% + paraaortic nodes »67% com Iliacs neg »73% Ipsi neg below IMA »69% pos above IMA *Mayo prospective accrual 1/2004 to 12/2006 6 Endometrioid Endometrial Cancer Role of Radiotherapy and Lymphadenectomy • Treatment paradigm shift • Minimize overtreatment –Identify pts not requiring LND and/or RT • Minimize undertreatment –Identify pts benefiting from LND and/or RT • Maximize outcomes Endometrioid Endometrial Cancer Role of Radiotherapy and Lymphadenectomy • Modality-based therapy • Radiotherapy vs. lymphadenectomy • Uterine histology • Disease-based therapy • Based on patterns of failure » Predicted by pathologic determinants • Selective Lymphadenectomy • Selective Radiotherapy • Selective Chemotherapy Endometrial Cancer Surgical Staging Role of Lymphadenectomy Karl Podratz MD PhD FACS 7