ORIGINAL ARTICLE Small Glandular Proliferations on Needle Biopsies Most Common Benign Mimickers of Prostatic Adenocarcinoma Sent in for Expert Second Opinion Mehsati Herawi, MD, PhD,* Anil V. Parwani, MD, PhD, Junji Irie, MD, and Jonathan I. Epstein, MD*†‡ Abstract: The current study aimed to determine the incidence of various benign mimickers of prostatic adenocarcinoma most commonly encountered in a busy consultation practice. All prostate needle biopsies from the consult service of one of the authors were prospectively evaluated over a 7-month period. Only cases with foci where the contributor questioned malignancy and which upon expert review the entire case was determined to be benign were included in this study. A total of 567 separate suspected atypical foci from 345 patients of a total of 4,046 patients (8.5%) received in consultation were identified. Of these, 281 foci (49.5%) had immunohistochemical (IHC) studies performed by the outside institution, which included high molecular weight cytokeratin (HMWCK) (n = 280), alphamethylacyl-CoA racemase (AMACR) (P504s) (n = 45), and p63 (n = 34). The most common mimicker was partial atrophy (203 of 567; 35.8%). Technically adequate IHC for basal cells was performed in 117 cases of partial atrophy with patchy or patchy/negative staining seen in 102 of 117 (87%), with the remaining 13% of cases completely negative. A total of 15 of 19 (79%) cases of partial atrophy were positive with AMACR. Crowded benign glands, insufficiently crowded or numerous to warrant a diagnosis of adenosis, was the second most common mimicker (146 of 567; 25.7%). Crowded benign glands had patchy or patchy/negative IHC for basal cells in 66 of 81 (81%) cases with the remaining 19% of cases completely negative. A total of 7 of 11 (64%) cases of crowded glands were positive for AMACR. In the past, complete atrophy, adenosis, seminal vesicle, and granulomatous prostatitis were considered common mimickers of prostate cancer on prostatic needle biopsies. Our study shows that currently partial atrophy and crowded benign glands are the most common benign changes causing diagnostic difficulty and prompting consultation. Negative or patchy staining for basal cells and positive staining for AMACR may contribute to diagnostic difficulty in these entities. From the Departments of *Pathology, †Urology, and ‡Oncology, Johns Hopkins University School of Medicine, Baltimore, MD. Dr. Parwani’s current affiliation is Department of Pathology, University of Pittsburgh, Pittsburgh, PA. Dr. Irie’s current affiliation is Department of Pathology, Nagasaki University School of Medicine, Nagasaki, Japan. Reprints: Jonathan I. Epstein, MD, Department of Pathology, 401 N. Broadway St., Rm 2242, Johns Hopkins Hospital, Baltimore, MD 21231 (e-mail: [email protected]). Copyright Ó 2005 by Lippincott Williams & Wilkins 874 Key Words: prostate cancer, mimickers, partial atrophy, adenosis, radiation atypia, inflammation, basal cell hyperplasia, basal cell markers, AMACR, second opinion (Am J Surg Pathol 2005;29:874–880) T he histopathologic interpretation of prostate needle biopsies remains the single most important tool for establishing a diagnosis of prostate cancer. The substantial increase in prostate needle biopsies in the prostate-specific antigen (PSA) screening era has not only resulted in the detection of very minute amounts of cancer but most likely has also resulted in an increased detection of small foci of a variety of benign lesions of the prostate. The distinction of benign small acinar proliferations from atypical acinar proliferations suspicious for cancer is crucial, since the subsequent clinical approach is different. Biopsies harboring a small focus of atypical glands frequently represent an undersampled cancer and a subsequent biopsy will show cancer in up to 50% of cases.1,6,16 In contrast, following a diagnosis of benign mimickers of cancer, such as atrophy or adenosis, a rebiopsy is usually not indicated. In this prospectively conducted study, we aimed to determine, based on the necessity for referral, which benign lesions of the prostate are currently causing the most diagnostic problems on needle biopsy for practicing pathologists in the United States. MATERIAL AND METHODS Case Selection Between January 28, 2004 and August 28, 2004, a total of 4,046 prostate needle biopsy cases were sent to one of the authors for consultation. Of these, 345 cases (8.5%) contained lesions of concern by the submitting pathologists that were diagnosed upon expert review as benign. Among the 345 cases, there were a total of 567 lesions of concern on different cores. These cases upon expert review did not have adenocarcinoma or glands suspicious for cancer or high-grade prostatic intraepithelial neoplasia (HGPIN) in any other cores. These cases originated from 183 centers (including commercial laboratories, community hospitals, and academic centers) in 35 different states. All cases had routine hematoxylin and Am J Surg Pathol Volume 29, Number 7, July 2005 Am J Surg Pathol Volume 29, Number 7, July 2005 Small Glandular Proliferations eosin slides available for evaluation. The mean age of men included in this study was 61.5 years (range, 33–88 years). Immunohistochemistry for HMWCK, p63, and AMACR For280 lesions, HMWCK-labeledsectionswere sent by the referring pathologist along with the hematoxylin and eosin slides for evaluation. In addition to HMWCK, stains for p63 were done on the outside on 34 cases; as the p63 staining results were virtually identical to those seen with HMWCK, p63 data will not be shown. The staining pattern for HMWCK was recorded as ‘‘patchy’’ when glands in a lesion reacted with antibodies for basal cells in a fragmented fashion, ‘‘patchy/negative’’ when the labeling of the basal cells was fragmented in some glands and completely absent in others within the lesion, ‘‘negative’’ when the entire lesion failed to reveal immunoreactive basal cells, and ‘‘positive’’ when all glands showed diffuse immunoreactivity for basal cells. Forty-five lesions had immunostains for AMACR performed at outside institutions. The AMACR staining was recorded ‘‘positive’’ or ‘‘negative.’’ Only staining stronger than that of the background benign glands was considered as positive. For the HMWCK and p63 stains, benign glands in the biopsy served as an internal positive control. Definitions of the Most Common Mimickers of Prostate Adenocarcinoma Partial atrophy is defined as prostate glands with scant apical cytoplasm that as a consequence of more abundant lateral cytoplasm do not appear basophilic at low magnification as compared with complete atrophy.11,24,29 Nuclei occupy almost the full cell height, and the cytoplasm is similar to the cytoplasm of the surrounding benign glands. Glands with complete atrophy have a very basophilic appearance at low magnification because of their overall scant cytoplasm and subsequent crowded nuclei.11 We do not subclassify complete atrophy into postatrophic hyperplasia and other patterns.2,8 By definition, most of our cases of complete atrophy would be considered by others as postatrophic hyperplasia as they consisted of crowded small atrophic glands mimicking cancer, as opposed to simple atrophy, which consists of more widely spaced larger atrophic glands which do not resemble prostate cancer. Adenosis is defined as a focus of very crowded small glands suspicious for cancer admixed with more recognizably benign glands with papillary infolding.4,11,13 Glands have pale to clear cytoplasm with nuclei showing a lack of very prominent nucleoli. ‘‘Crowded glands’’ is defined as a focus of benign crowded glands that unlike adenosis do not as closely mimic adenocarcinoma of the prostate.11 ‘‘Benign glands’’ was used when a focus was marked by the contributing pathologist as atypical yet did not resemble prostate cancer either architecturally or cytologically upon expert review. RESULTS Of 4,046 prostate needle biopsies received in consultation in a period of 7 months, 345 cases (8.5%) contained 567 lesions of concern by the submitting pathologists that upon expert review were identified as benign. q 2005 Lippincott Williams & Wilkins FIGURE 1. Partial atrophy. Group of cystically dilated partially atrophic glands of various sizes with mildly enlarged separated nuclei. Scattered basal cells are noticeable. Partial atrophy (Figs. 1–3) was the most common mimicker of cancer identified in our study population seen in 35.8% of lesions flagged as atypical by the outside pathologist (Table 1). The results of immunohistochemical staining performed at the referring institutions are depicted in Table 2. Of note, 87% of these cases showed either patchy or patchy/negative staining for HMWCK with 13% of the lesions totally negative, and no case with diffuse positivity. In contrast, complete atrophy (Fig. 4), the fourth most common benign mimicker of cancer, had in 37.5% of the lesions diffuse positivity for HMWCK (Table 2). AMACR labeled 79% of partial atrophy lesions. ‘‘Crowded glands’’ (Figs. 5–7) was the second most common benign mimicker of cancer identified in 25.7% of the lesions with similar immunohistochemical results to partial atrophy (Table 2). ‘‘Benign glands’’ (Fig. 8), the third most common lesion sent in for consultation, which appeared totally normal upon expert review, had patchy, patchy/negative, or negative staining for HMWCK in 62.5% of the lesions. Radiation atypia, inflammatory atypia, adenosis (Fig. 9), and basal cell hyperplasia (Fig. 10) comprised the fifth through eighth next most commonly seen benign mimickers (Table 1). Inflammatory atypia consisted of glands showing atypia due to acute (14 of 26), chronic (4 of 26), or mixed acute/chronic (8 of 26) inflammation; immunohistochemistry done at outside institutions was only performed on cases with acute inflammation. The incidence of the other less frequent benign lesions sent in for consultation and their immunohistochemical results are listed in Tables 1 and 2. Miscellaneous lesions consisted of cribriform hyperplasia (n = 2), urothelial metaplasia (n = 2), mucinous metaplasia (n = 2), nonspecific granulomatous prostatitis (n = 2), small benign glands with pale cytoplasm (n = 2), infarct (n = 1) (Fig. 11), paraganglioma (n = 1), colonic mucosa (n = 1), and a tangential cut through basal cells (n = 1).3,11,14,23,25–27 875 Am J Surg Pathol Volume 29, Number 7, July 2005 Herawi et al FIGURE 2. Partial atrophy. A, Disorganized partially atrophic glands with scant cytoplasm and enlarged nuclei containing small nucleoli. B, Staining with p63 is patchy and negative. DISCUSSION The differential diagnosis of small acinar proliferations in needle biopsies of the prostate is broad and includes prostate cancer and various benign mimickers of prostate cancer.15,17,31,32 Lesions cited as mimicking prostate cancer have changed over the years. Whereas seminal vesicle tissue had in the distant past been considered one of the common mimickers of prostate cancer,18,30–32 in the current study there was not a single case of seminal vesicle tissue that was sent in for consultation as a result of diagnostic difficulties. In a study published from our institution in 1995, complete atrophy and adenosis were the most common benign lesions sent in for consultation.12 That basal cell hyperplasia, cribriform hyperplasia, and infarcts were only infrequently seen in the current study is not surprising as these lesions preferentially affect the transition zone and are more frequently encountered on transurethral resections.3,9,11,23 Radiation atypia, although seen in only 5.6% of cases, was the fifth most common referred lesion.5,7,22 As radiation continues to grow in popularity as a treatment of early prostate cancer and patients treated in the 876 FIGURE 3. Partial atrophy. A, Haphazardly arranged small to medium-sized glands with partial atrophy. B, Glands stained with a cocktail containing antibodies to p63, HMWCK and AMACR showing positive staining with AMACR and a variably fragmented and absent basal cell layer with p63 and HMWCK. past are followed for longer intervals where they will be at risk for recurrence, radiation atypia can be expected in the future to account for a greater proportion of benign cases referred in for consultation. TABLE 1. Incidence of Lesions Diagnosis Partial atrophy Crowded glands ‘‘Benign glands’’ Complete atrophy Radiation atypia Inflammatory atypia Adenosis (atypical adenomatous hyperplasia) Basal cell hyperplasia Miscellaneous No. % of Lesions 203/567 146/567 56/567 55/567 32/567 26/567 (35.8) (25.7) (9.8) (9.7) (5.6) (4.6) 21/567 (3.7) 14/567 (2.5) 14/567 (2.5) q 2005 Lippincott Williams & Wilkins Am J Surg Pathol Volume 29, Number 7, July 2005 Small Glandular Proliferations TABLE 2. Immunohistochemical Results. No. (%) HMWCK Diagnosis Partial atrophy Crowded glands ‘‘Benign glands’’ Complete atrophy Radiation atypia Inflammatory atypia Adenosis (AAH) Basal cell Hyperplasia Patchy 45/117 30/80 6/16 6/16 (38) (37) (37.5) (37.5) 2/3 (66.7) 5/11 (45.4) Patchy Negative 57/117 36/80 2/16 2/16 (49) (450) (12.5) (12.5) 1/3 (33.3) 4/11 (36.4) The most common benign lesion that caused difficulty for pathologists was partial atrophy.24,29 In part, difficulty with this entity can be attributed to its relatively recent description. Although partial atrophy was first described in 1992,10 it was FIGURE 4. Complete atrophy. A, Haphazardly arranged small glands with complete atrophy with a pseudo-infiltrative growth pattern. B, High-power of atrophic glands with shrunken cytoplasm resulting in high nuclear to cytoplasmic ratio and basophilic appearance. q 2005 Lippincott Williams & Wilkins Negative 15/117 14/80 2/16 2/16 (13) (17) (12.5) (12.5) Positive 0/117 3/80 6/16 6/16 5/5/ (0) (4) (37.5) (37.5) (100) 2/11 (18.2) AMACR Positive 15/19 (79) 7/11 (67) 1/4 (25) 3/3 (100) first reported in the periodic literature only in 1998.24 Partial atrophy mimics cancer in several aspects. Architecturally, partial atrophy consists of crowded glands often with a disorganized growth pattern. In contrast to fully developed atrophy, which can typically be diagnosed at scanning magnification owing to the presence of well-formed glands with a very basophilic appearance, partial atrophy has pale cytoplasm lateral to the nuclei giving rise to pale staining glands that more closely mimic cancer. The nuclei in partial atrophy are slightly enlarged and irregular with occasional visible nucleoli, and basal cells may be difficult to identify on routine stained sections, all contributing to the difficulty in distinguishing this lesion from cancer. Clues for its correct diagnosis include scant apical cytoplasm with nuclei extending to the full cell height, pale lateral cytoplasm similar to the cytoplasm seen in adjacent more recognizable benign glands, luminal undulations as contrasted to the straight luminal borders often seen with malignant glands, and association with regular atrophy. An additional factor that contributes to the difficulty in distinguishing cancer from partial atrophy is that this mimicker may stain negative with markers for basal cells and stain positive with AMACR. In our study population, HMWCK performed in 117 cases of partial atrophy showed FIGURE 5. Crowded glands. Focus of crowded benign appearing glands insufficient to establish a diagnosis of adenosis. 877 Herawi et al Am J Surg Pathol Volume 29, Number 7, July 2005 FIGURE 6. Collection of benign crowded glands. patchy or patchy/negative staining in 87% of cases, with the remaining 13% cases completely negative. Foci of partial atrophy were positive with AMACR in 79% of cases where stains were performed by the outside institution. The current study is the only one that we are aware of that reports on the basal cell and AMACR immunohistochemical staining pattern in a series of partial atrophy. However, our cases of partial atrophy are selected, in that only cases of partial atrophy sent for consultation were studied and not all cases of partial atrophy sent for review had immunohistochemical stains. The second most common benign lesion misinterpreted as cancer was ‘‘crowded glands.’’ Foci of crowded glands are defined as benign glands insufficiently crowded or numerous to warrant a diagnosis of adenosis.11 As with partial atrophy, crowded glands in our study showed on immunostains a patchy or patchy/negative labeling for basal cells in 81% of cases, FIGURE 7. Crowded glands. Focus of minimally crowded small glands. The small glands show identical nuclear and cytoplasmic features as the adjacent larger benign glands with papillary infolding. 878 FIGURE 8. Benign glands. Benign-appearing glands without cytologic atypia. FIGURE 9. Adenosis. A, Adenosis consisting of a collection of closed-packed glands of different sizes with numerous corpora amylacea. B, HMWCK labels the basal cells focally in a patchy fashion; the majority of glands show negative staining. q 2005 Lippincott Williams & Wilkins Am J Surg Pathol Volume 29, Number 7, July 2005 FIGURE 10. Basal cell hyperplasia. Small nucleoli may be visible in cases of basal cell hyperplasia. with the remaining 19% of cases completely negative. A majority of cases of crowded glands (64%) were positive for AMACR. In a similar vein, ‘‘benign glands’’ was the third most common benign lesion confused with carcinoma. Although these foci on the routinely stained sections did not resemble cancer, 62.5% of these cases demonstrated patchy, patchy/negative, or negative staining with HMWCK. This immunohistochemical profile causes confusion with carcinoma and was undoubtedly one of the factors leading to these cases being referred for consultation. That these lesions are not well-defined entities where a pathologist can assign them a name, look them up in a book, and read about them, also most likely contributed to the difficulty in recognizing them as benign. While the use of immunohistochemistry for basal cells and more recently AMACR has aided in the diagnosis of prostate cancer, there have also been pitfalls with their use. Small Glandular Proliferations Problems with the use of these markers were highlighted with the three most common benign mimickers of cancer seen on needle biopsy in the current study. Our work emphasizes that pathologists must ultimately establish their diagnosis on the hematoxylin and eosin-stained sections using immunohistochemistry as an adjunct. Pathologists must be educated on the subtle features of partial atrophy. Patchy staining in a lesion is diagnostic of a benign process, as long as the negatively stained glands have the same hematoxylin and eosin morphology as those glands where basal cells are documented. Even entirely negative staining for basal cells in a small focus of glands can be entirely compatible with a benign process. AMACR has been touted as being almost entirely specific for cancer, with the exception of labeling HGPIN and some cases of adenosis.21,28 The percentage of positive staining with AMACR in benign glands has been reported in previous studies to be 8% to 12%.19,20 In the current study, both partial atrophy and crowded benign glands sent in for consultation were positive in 79% and 67% of cases, respectively. The percentage of these cases staining for AMACR is most likely not representative of what would be seen in these lesions as a whole. Rather, some of these cases were most likely sent in for consultation as a result of the AMACR positivity. Furthermore, we did not distinguish between weak, moderate, and strong staining and accepted any staining stronger than the background as positive. Another potential explanation for the differences between our study and those citing a lower incidence of AMACR positivity in benign glands is that our cases were stained in a variety of institutions with varying techniques and antibodies. Nonetheless, our study demonstrates how AMACR staining can give misleading results. In addition to highlighting the potential pitfalls with immunohistochemical staining for HMWCK and AMACR, the current study directs where greater educational efforts are needed to improve the accuracy in diagnosing prostate cancer. REFERENCES FIGURE 11. Infarct. High power view of prostate glands immediately adjacent to an infarct showing reactive immature squamous metaplasia with enlarged hyperchromatic nuclei. q 2005 Lippincott Williams & Wilkins 1. Allen EA, Kahane H, Epstein JI. Repeat biopsy strategies for men with atypical diagnoses on initial prostate needle biopsy. Urology. 1998;52:803–807. 2. Amin MB, Tamboli P, Varma M, et al. 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