The 1000 Mitoses Project: A Consensus-Based International Collaborative Study on Mitotic Figures Classification.

Introduction. The identification of mitotic figures is essential for the diagnosis, grading, and classification of various different tumors. Despite its importance, there is a paucity of literature reporting the consistency in interpreting mitotic figures among pathologists. This study leverages publicly accessible datasets and social media to recruit an international group of pathologists to score an image database of more than 1000 mitotic figures collectively. Materials and Methods. Pathologists were instructed to randomly select a digital slide from The Cancer Genome Atlas (TCGA) datasets and annotate 10-20 mitotic figures within a 2 mm2 area. The first 1010 submitted mitotic figures were used to create an image dataset, with each figure transformed into an individual tile at 40x magnification. The dataset was redistributed to all pathologists to review and determine whether each tile constituted a mitotic figure. Results. Overall pathologists had a median agreement rate of 80.2% (range 42.0%-95.7%). Individual mitotic figure tiles had a median agreement rate of 87.1% and a fair inter-rater agreement across all tiles (kappa = 0.284). Mitotic figures in prometaphase had lower percentage agreement rates compared to other phases of mitosis. Conclusion. This dataset stands as the largest international consensus study for mitotic figures to date and can be utilized as a training set for future studies. The agreement range reflects a spectrum of criteria that pathologists use to decide what constitutes a mitotic figure, which may have potential implications in tumor diagnostics and clinical management.

Ancient Schwannoma of the Thigh: Metaplastic Ossification, Cartilage Formation, and Extensive Calcification.

Benign soft tissue masses that present with atypical features on imaging may erroneously be diagnosed as malignant processes. An ancient schwannoma, a schwannoma variant with pronounced degenerative features, is one possible etiology of an incidental soft tissue tumor. This case report describes a 69-year-old male with a history of lung carcinosarcoma who presented to the orthopedic oncology office following an incidental positron emission tomography-computed tomography (PET-CT) finding of a posterior, lateral thigh mass with extensive calcifications. Subsequent excision and pathological analysis revealed an ancient schwannoma with advanced degenerative features, including metaplastic bone and cartilage formation. Various degenerative changes may typically be identified with pathological analysis. In addition to the degenerative findings on pathological analysis, our case highlights an atypical instance where extensive calcification and ossification are evident radiographically. This case emphasizes the importance of considering ancient schwannoma when a calcified soft tissue mass is encountered, in addition to the list of more common calcified soft tissue masses.

Transformed Testicular Teratoma to Embryonic-Type Neuroectodermal Tumor With Metastasis to Mediastinum.

Testicular teratomas may present in both prepubertal and adult men; however, the prognosis differs greatly between these 2 populations. In children, teratomas (prepubertal type) most often occur before the age of 4, are generally seen in their pure form, and behave in a benign fashion. In adults (postpubertal type), teratomas are usually part of a mixed germ cell tumor, and they have the potential to be found at metastatic sites, especially following chemotherapy for non-teratomatous germ cell tumor. Analyses of metastases from germ cell tumors and teratomas from the same patient have demonstrated a high degree of concordance in the observed genetic abnormalities. In rare cases, testicular teratoma can transform into a malignant germ cell tumor. One such type of transformation is into a primitive neuroectodermal tumor. These tumors are malignant and often metastasize to the retroperitoneum but may also metastasize to other sites. A multimodal treatment approach is needed, including surgery and adjuvant chemotherapy. We describe a rare case of malignant transformation of a testicular teratoma into a primitive neuroectodermal tumor with metastasis to the mediastinum. The patient was treated with radical orchiectomy, retroperitoneal lymph node dissection, and adjuvant vincristine, adriamycin, and cyclofosfamide alternating with ifosfamide and etoposide (VAC/IE therapy).

A Case Report of Progressive Paget Disease Mimicking a Malignancy: A Second Episode 21 Years Later.

Paget osteosarcoma is a rare but serious complication of Paget disease requiring immediate management before further malignant transformation. This case report examines the progression of a previously reported case of Paget disease with atypical pseudotumor manifestation, mimicking osteosarcoma over a 21-year time lapse. After presenting with substantial pain and elevated alkaline phosphatase levels, imaging proved extensive bony expansion of the lesion with high-grade trabecular and cortical thickening and extraosseous soft-tissue extension, prompting the need for biopsy to rule out Paget sarcoma. The atypical features of the pseudotumor's development helps distinguish key radiographic and clinical criteria for malignant development.

Testicular Juvenile Granulosa Cell Tumors Demonstrate Recurrent Loss of Chromosome 10 and Absence of Molecular Alterations Described in Ovarian Counterparts.

Testicular juvenile granulosa cell tumors (JGCTs) are a rare type of sex cord-stromal tumor, accounting for <5% of all neoplasms of the prepubertal testis. Previous reports have demonstrated sex chromosome anomalies in a small subset of cases, but the molecular alterations associated with JGCTs remain largely undescribed. We evaluated 18 JGCTs using massive parallel DNA and RNA sequencing panels. The median patient age was <1 month (range, newborn to 5 months). The patients presented with scrotal or intra-abdominal masses/enlargement, and all underwent radical orchiectomy (17 unilateral and 1 bilateral). The median tumor size was 1.8 cm (range, 1.3-10.5 cm). Histologically, the tumors were purely cystic/follicular or mixed (ie, solid and cystic/follicular). All cases were predominantly epithelioid, with 2 exhibiting prominent spindle cell components. Nuclear atypia was mild or absent, and the median number of mitoses was 0.4/mm2 (range, 0-10/mm2). Tumors frequently expressed SF-1 (11/12 cases, 92%), inhibin (6/7 cases, 86%), calretinin (3/4 cases, 75%), and keratins (2/4 cases, 50%). Single-nucleotide variant analysis demonstrated the absence of recurrent mutations. RNA sequencing did not detect gene fusions in 3 cases that were sequenced successfully. Recurrent monosomy 10 was identified in 8 of 14 cases (57%) with interpretable copy number variant data, and multiple whole-chromosome gains were present in the 2 cases with significant spindle cell components. This study demonstrated that testicular JGCTs harbor recurrent loss of chromosome 10 and lack the GNAS and AKT1 variants described in their ovarian counterparts.

Clear cell mesotheliomas with inactivating VHL mutations and near-haploid genomic features.

Clear cell mesothelioma is uncommon and shows predominance of clear cells with resemblance to clear cell carcinomas. Clinicopathologic and molecular descriptions of clear cell mesothelioma remained limited. In this study, we identified an index patient with clear cell mesothelioma, confirmed by immunohistochemical and ultrastructural studies. Targeted next-generation sequencing revealed the presence of an inactivating VHL mutation. We then systematically searched for VHL-mutant mesotheliomas in a comprehensive genomic profiling database of 1532 mesotheliomas. Collectively, we identified a cohort of four VHL-mutant clear cell mesotheliomas, including three peritoneal and one pleural tumors from three females and one male, with age range of 47-68 (median 63) years. Histologically, each tumor showed a microcystic to tubulopapillary architecture with prominent clear cells. By next-generation DNA sequencing, each of the four clear cell mesotheliomas harbored inactivating VHL mutations, while lacking other alterations typical of mesotheliomas such as BAP1, NF2, SETD2, CDKN2A, CDKN2B, TP53, and PTEN. By using low-pass whole genome sequencing on the index case and targeted next-generation sequencing on the remaining three cases, we identified extensive loss of heterozygosity throughout the genome but consistently sparing chromosomes 5, 7, and 20, characteristic of genomic near-haploidization. In summary, clear cell mesotheliomas were characterized by inactivating VHL mutations and genomic near-haploidization and appeared to represent a distinct clinicopathologic and molecular category of mesotheliomas. Our findings implicate VHL in the pathogenesis of a subset of mesotheliomas, particularly those with clear cell morphology.

Low-Grade Oncocytic Tumor of Kidney (CK7-Positive, CD117-Negative): Incidence in a single institutional experience with clinicopathological and molecular characteristics.

Low-grade oncocytic tumor of the kidney (LOT) is characterized by cytoplasmic eosinophilia and a CK7-positive/CD117-negative immunophenotype. Morphologically, they exhibit overlapping features with oncocytoma and chromophobe renal cell carcinoma. Our aim was to obtain long-term clinical follow-up data, clinicopathological and molecular characteristics, and incidence of LOT. Tissue microarrays were constructed from 574 tumors historically diagnosed as oncocytoma and surgically treated at Mayo Clinic between 1970 and 2012, and immunostained for CK7 and CD117. An extended immunophenotype was obtained on whole slide sections, along with FISH for CCND1 rearrangement status and chromosomal microarray for copy number status. In addition, two cases were retrospectively identified in a set of tuberous sclerosis complex (TSC)-associated neoplasms and three more cases diagnosed on needle core biopsies were obtained during routine clinical practice. Twenty-four cases of LOT were identified among 574 consecutive tumors diagnosed as oncocytoma and treated with partial or radical nephrectomy, corresponding to an incidence of 4.18% of tumors historically diagnosed as oncocytomas, and 0.35% of 6944 nephrectomies performed between 1970 and 2012. Overall, 29 cases of LOT were identified in three clinical settings: sporadic, TSC-associated, and end-stage renal disease (ESRD). Multifocality was seen only in the setting of TSC and ESRD. No metastases attributable to LOT were identified (median follow-up 9.6 years). There were no recurrent arm level copy number changes detected by chromosomal microarray and all tested cases were negative for CCND1 rearrangement by FISH. LOT is an uncommon eosinophilic renal neoplasm with an indolent prognosis that constitutes ∼4% of tumors historically diagnosed as oncocytoma. The morphologic, immunophenotypic, and molecular features of this neoplasm suggest it is a distinct entity of renal neoplasia.

Growth and differentiation factor 15 and NF-κB expression in benign prostatic biopsies and risk of subsequent prostate cancer detection.

Growth and differentiation factor 15 (GDF-15), also known as macrophage inhibitory cytokine 1 (MIC-1), may act as both a tumor suppressor and promotor and, by regulating NF-κB and macrophage signaling, promote early prostate carcinogenesis. To determine whether expression of these two inflammation-related proteins affect prostate cancer susceptibility, dual immunostaining of benign prostate biopsies for GDF-15 and NF-κB was done in a study of 503 case-control pairs matched on date, age, and race, nested within a historical cohort of 10,478 men. GDF-15 and NF-κB expression levels were positively correlated (r = 0.39; p < 0.0001), and both were significantly lower in African American (AA) compared with White men. In adjusted models that included both markers, the odds ratio (OR) for NF-κB expression was statistically significant, OR =0.87; p = 0.03; 95% confidence interval (CI) =0.77-0.99, while GDF-15 expression was associated with a nominally increased risk, OR =1.06; p = 0.27; 95% CI =0.96-1.17. When modeling expression levels by quartiles, the highest quartile of NF-κB expression was associated with almost a fifty percent reduction in prostate cancer risk (OR =0.51; p = 0.03; 95% CI =0.29-0.92). In stratified models, NF-κB had the strongest negative association with prostate cancer in non-aggressive cases (p = 0.03), older men (p = 0.03), and in case-control pairs with longer follow-up (p = 0.02). Risk associated with GDF-15 expression was best fit using nonlinear regression modeling where both first (p = 0.02) and second (p = 0.03) order GDF-15 risk terms were associated with significantly increased risk. This modeling approach also revealed significantly increased risk associated with GDF-15 expression for subsamples defined by AA race, aggressive disease, younger age, and in case-control pairs with longer follow-up. Therefore, although positively correlated in benign prostatic biopsies, NF-κB and GDF-15 expression appear to exert opposite effects on risk of prostate tumor development.

Microglandular Adenosis and Associated Invasive Carcinoma.

CONTEXT.­: Microglandular adenosis is a rare borderline neoplastic lesion of the breast composed of haphazardly located small, round tubules with a single cell layer interspersed within breast stroma and/or adipose tissue. Microglandular adenosis is devoid of a myoepithelial cell layer, and has a characteristic immunophenotype, being positive for S100 and negative for estrogen receptor, progesterone receptor, and HER2/neu. When associated with cancer, microglandular adenosis and associated invasive carcinoma share the same molecular alterations, including TP53 mutation; therefore, microglandular adenosis is considered a nonobligate precursor of triple (HER2/neu, estrogen and progesterone receptors)-negative breast carcinoma. Microglandular adenosis is an important diagnostic pitfall as it can be easily mistaken for a low-grade invasive carcinoma. OBJECTIVE.­: To provide a review of the clinicopathologic features of microglandular adenosis and associated invasive carcinoma, with emphasis on key features separating entities in the differential diagnosis. DATA SOURCES.­: Review of current literature on microglandular adenosis and associated invasive carcinoma and personal experience of authors. CONCLUSIONS.­: Microglandular adenosis can mimic breast carcinoma; attention to key features, including morphologic-immunophenotypic correlation, is essential in establishing the diagnosis.

KIF3B protein expression loss correlates with metastatic ability of prostate cancer.

Kinesin family member 3B (KIF3B) is a microtubule motor kinesin involved in mitotic progression and vasculotropism. A novel therapeutic target, it is overexpressed in several cancers [PMID 29904055]. Its significance in prostate cancer (PC) was uncertain. METHODS: 89 cases, including tissue microarrays from 70 prostatectomies comprising matched cancer and benign spots, 19 additional prostatectomy tissues, plus 16 prostate cancer metastases (7 nodal and 9 distant sites; 8 had matched primary PC) were stained with rabbit polyclonal KIF3B antibody. Cytoplasmic immunoreactivity was scored: 0 (negative) to 3+ (strong and diffuse). 39 patients had no nodal metastases, 31 had positive lymph nodes, and 19 had nodes not sampled. Gleason grade groups were 1 (9), 2 (28), 3 (39), 4 (1), and 5 (12). 15 cases had cribriform pattern. AJCC stages were 2 (48), 3 (29), unknown (12). RESULTS: KIF3B in PC (mean 1.0) was higher than in benign prostate (mean 0.1, P<0.01, Student t-test). All 7 available nodal metastases of PC were negative. One-third of primary PCs with nodal metastases lost all expression, compared to retained expression in all but one PC without nodal metastasis (P<0.01, chi-square). The former group also had stronger staining (mean 1.0) than metastases (mean 0.3) (P<0.01, Student t-test) and had fewer cases with any positive (>0) expression compared to cases without metastases or with unsampled lymph nodes (P<0.01, chi-square test). Reactivity of paired metastatic tissue and primary PC correlated strongly (Pearson coefficient: +0.7). No significant trends were found by grade group, cribriform status, or stage. CONCLUSIONS: KIF3B is a PC marker. Metastatic cancers showed less KIF3B expression than their primary PC counterparts, and primary cases with positive nodes demonstrated reduced positivity, suggesting use as a prognostic marker. It is possible that KIF3B protein becomes altered prior to metastases, preventing immunohistochemical detection.

Proteinaceous Lymphadenopathy in a Young Patient With History of Classical Hodgkin Lymphoma: A Case Report With Literature Review.

Proteinaceous lymphadenopathy (PLD) is a poorly defined, underreported pathological entity of uncertain etiology characterized by massive deposition of amorphous, eosinophilic, and periodic acid-Schiff-positive material involving lymph nodes, which is distinct from amyloid and clonal immunoglobulin deposition. PLD can resemble collagen sclerosis and needs to be differentiated from lymphomas with sclerosis, particularly classical Hodgkin lymphoma, nodular sclerosis type, and therefore is an important pitfall in the diagnosis of lymphoma with sclerosis. We are reporting a young patient with history of classical Hodgkin lymphoma who eventually developed PLD and review the literature on this subject.

Secretory carcinoma of the parotid with adenoid cystic carcinoma cytological pattern: A cytological-pathological correlation with literature review.

Secretory carcinoma (SC) is a rare low-grade malignant tumor, defined by ETV6-NTRK3 fusion, identifiable by FISH. We describe a case in a 58-year-old male with a painless slowly growing 16mm palpable mass within left superficial parotid. FNA of the mass showed highly cellular specimen with moderate to large pleomorphic cells with round to ovoid nuclei with vesicular chromatin and distinct nucleoli. Cells had moderate to large amounts of vacuolated cytoplasm. Abundant globular metachromatic material, resembling that of adenoid cystic carcinoma, was noted. This material was seen extracellularly and intracytoplasmic, and stained magenta on Diff-Quik and blue-green on Papanicolaou-stained slides. The tumor cells on a cell block preparation were positive for Mammaglobin and S-100. PAS stain highlighted extracellular and intracytoplasmic secretions. FNA diagnosis was "Positive for Malignancy. Morphologic features most compatible with Mammary Analogue Secretory Carcinoma". ETV6 FISH studies as well as histologic examination of excised tumor confirmed the diagnosis. Finding the globular metachromatic material in SC, that is generally seen in adenoid cystic carcinoma, broadens a cytological differential diagnosis of both entities. Cytological differential diagnosis, clinical, histological, immunohistochemical, and molecular features of secretory carcinomas are discussed in this study.

PTEN loss and p27 loss differ among morphologic patterns of prostate cancer, including cribriform.

The presence and extent of cribriform pattern of prostate cancer portend recurrence and cancer death. The relative expressions within this morphology of the prognostically adverse loss of PTEN, and the downstream inactivation of cell cycle inhibitor p27/Kip1 had been uncertain. In this study, we examined 52 cases of cribriform cancer by immunohistochemistry for PTEN, p27, and CD44 variant (v)7/8, and a subset of 17 cases by chromogenic in situ hybridization (ISH) using probes for PTEN or CDKN1B (gene for p27). The fractions of epithelial pixels positive by immunohistochemistry and ISH were digitally assessed for benign acini, high-grade prostatic intraepithelial neoplasia, and 8 morphologic patterns of cancer. Immunostaining results demonstrated that (1) PTEN loss was significant for fused small acini, cribriform-central cells, small cribriform acini, and Gleason grade 5 cells in comparison with other acini; (2) p27 loss was significant only for cribriform-peripheral cells and borderline significant for fused small acini in comparison with benign acini; and (3) CD44v7/8 showed expression loss in cribriform-peripheral cells; other comparisons were not significant. ISH showed that cribriform cancer had significant PTEN loss normalized to benign acini (P<.02), whereas Gleason 3 cancer or fused small acini did not. With CDKN1B, the degree of signal loss among various cancer morphologies was insignificant. In conclusion, molecular disparities emerged between the fused small acini and cribriform patterns of Gleason 4 cancer. PTEN or p27 loss as prognostic factors demands distinct assessment in the varieties of Gleason 4 cancer, and in the biphenotypic peripheral versus central populations in cribriform structures.

Myoepithelioma of soft tissue: A cytological-pathological correlation with literature review.

Myoepitheliomas of soft tissue are rare tumors with variable morphologic, immunohistochemical and molecular profiles and therefore are diagnostically challenging for pathologists. We report a case in a 60-year-old male with a painless slowly growing 3cm mass on left medial forefoot. Core biopsy of the mass showed a neoplastic proliferation of plasmacytoid tumor cells, consistent with myoepithelioma of soft tissue. Immunohistochemical stains demonstrated positivity of the tumor cells for cytokeratin AE1/AE3, CK18, S-100 protein and myosin heavy chain (SMMS-1), supporting the diagnosis. Fine-needle aspiration was performed intraoperatively before the resection of the mass. Air-dried Diff-Quik stained cytology slides showed singly scattered and loosely cohesive clusters of plasmacytoid and spindle cells with dense basophilic cytoplasm, distinct cytoplasmic borders, and round to oval mildly pleomorphic nuclei with smooth nuclear membrane. Scattered naked nuclei, binucleated tumor cells, as well as tumor cells with wispy elongated cytoplasm were also seen. Occasional clusters of cells were intimately associated with metachromatic fibrillary stromal material. Histologic examination of the resected tumor confirmed the diagnosis of myoepithelioma. Molecular studies showed absence of EWSR1 rearrangement. Cytological differential diagnosis, clinical, histological, immunohistochemical, and molecular features of soft tissue myoepitheliomas are discussed in this study.