Ann Boguniewicz

Clinical next‐generation sequencing successfully applied to fine‐needle aspirations of pulmonary and pancreatic neoplasms

Next‐generation sequencing was performed on pulmonary and pancreatic fine‐needle aspirations (FNAs) and on paired FNAs and resected primary tumors from the same patient.

DNA ploidy and cell cycle analysis in breast cancer.

During the past 10 years there has been considerable interest in the application of new technologies to identify human malignancy and predict disease outcome. Markers of cell proliferation and the technologies of flow cytometry and image analysis for the determination of DNA total content in human tumor cells have been studied in breast cancer for 20 years. In this review, the uses and limitations of these technologies for the determination of ploidy status are discussed. This review also considers the prognostic significance and potential clinical utility of ploidy measurements, S phase calculation, and individual cell cycle regulatory biomarker expression levels.

Overexpression of WWP1 is associated with the estrogen receptor and insulin-like growth factor receptor 1 in breast carcinoma.

WWP1, a HECT type E3 ubiquitin ligase frequently amplified and overexpressed in breast cancer, has the potential to become a useful clinical biomarker and therapeutic target in breast cancer. Here, we performed immunohistochemical staining in formalin‐fixed and paraffin‐embedded tissue sections from 187 cases of primary invasive mammary carcinoma [137 ductal carcinomas (IDC) and 50 lobular carcinomas (ILC)] by using a monoclonal anti‐WWP1 antibody. The normal breast epithelium and adjacent benign epithelium are essentially negative for WWP1. Cytoplasmic WWP1 immunoreactivity was observed in 76/187 (40.6%) tumors and showed a positive correlation with ERα (p = 0.05) and IGF‐1R proteins (p = 0.001) in this cohort. The positive correlations between WWP1 and ER/IGF‐1R were also observed in a panel of 12 breast cancer cell lines by Western blot. Interestingly, the ER levels are decreased when WWP1 is silenced in ER positive MCF7 and T47D breast cancer cell lines. Finally, WWP1 ablation collectively inhibits cell proliferation with tamoxifen in MCF7 and T47D, as measured by 3H‐thymidine incorporation assays. These findings suggest that WWP1 may play an important role in ER positive breast cancer.

Breast Cancer in a Man With Human Immunodeficiency Virus Infection

Non-acquired immunodeficiency syndrome (AIDS)-defining neoplasms are being increasingly recognized in patients infected with the human immunodeficiency virus (HIV). The incidence of Hodgkins disease and seminoma has recently been reported to be increasing in these patients. This article describes the second case of breast cancer in an HIV-infected male patient. A total of 11 cases of coincident breast cancer and HIV infection have previously been reported. It may be prudent to consider breast cancer in the differential diagnosis of an axillary mass in an HIV-infected patient.

Pigmented villonodular synovitis : Report of a case with diagnostic synovial fluid cytologic features

BACKGROUND The identification of neoplastic cells in synovial fluid is an uncommon occurrence and most often is related to extension of extraarticular tumor into the joint space than to primary neoplasm arising in the joint. CASE In this report the cytologic features of synovial fluid obtained from the right knee of an 18-year-old male with biopsy-proven pigmented villonodular synovitis are described and compared with the features of the concurrent surgical specimen. CONCLUSION The cytologic features of pigmented villonodular synovitis in synovial fluid include abundant mononuclear histiocytic cells occurring singly and in papillary clusters, hemosiderin within histiocytes and few multinucleated giant cells.

Adrenal pseudocysts: Evidence of their posthemorrhagic nature

Hemorrhagic adrenal pseudocysts are uncommon nonneoplastic lesions that have been reported as secondary to intraparenchymal hemorrhage or alternatively related to endothelial (vascular) cysts. Ultrastructural and immunohistochemical evidence in support of the latter has been presented, but the exact nature of hemorrhagic adrenal pseudocysts remains poorly defined. We evaluated six surgical specimens of hemorrhagic adrenal pseudocysts using immunohistochemical staining for CD31 and CD34, as well as conventional histochemistry. All six cases had hemorrhagic contents within a wall of variable thickness possessing focal areas of linear, disrupted elastin, and smooth muscle. Three cases demonstrated extensive thrombosis with organization, including papillary endothelial hyperplasia, simulating angiosarcoma. In these cases, CD31 and CD34 staining decorated areas of papillary endothelial hyperplasia as well as foci of the internal cyst lining, whereas the other cases were negative for both antibodies. Of interest is the history of FNA prior to surgical resection in three cases of hemorrhagic adrenal pseudocysts, two of which showed papillary endothelial hyperplasia. The presence of papillary endothelial hyperplasia and our immunohistochemical findings support, the conclusion that adrenal pseudocysts are posthemorrhagic and derive from vascular disruption. Furthermore, FNA or other interventional studies may be associated with papillary endothelial hyperplasia in hemorrhagic adrenal pseudocysts.

PD-L1 protein expression in tumour cells and immune cells in mismatch repair protein-deficient and -proficient colorectal cancer: the foundation study using the SP142 antibody and whole section immunohistochemistry

Aims Routine application of PD-L1 immunohistochemistry (IHC) in colorectal cancer (CRC) is limited due to lack of standardized scoring criteria, antibody clones, and intratumoral staining heterogeneity. We assessed PD-L1 protein expression on full face CRC tissue sections and applied two algorithms based on the published clinical trials that support the recent FDA approval for immune checkpoint inhibitors (ICPI) therapy in non-small cell lung cancer (NSCLC). Methods PD-L1/CD274 IHC (Roche/Ventana, clone SP142) was performed on representative tumour blocks from 52 mismatch repair-deficient (MMR-D) and 52 MMR-proficient (MMR-P) CRCs. Membranous PD-L1 expression was scored for the tumour cell (TC) and tumour-infiltrating immune cell (IC) components. PD-L1 positivity status was determined based on the published NSCLC clinical trials that utilized the Ventana SP142 assay. Hybrid capture-based comprehensive genomic profiling (CGP) was performed on a separate set of 2268 clinically advanced CRCs and the frequency of PD-L1/PD-L2 amplification was determined. Results PD-L1 expression in the TC and IC correlated with MMR-D (p=0.013, p<0.0001), T stage (p=0.036, p=0.0036) and clinical stage (p=0.022, p=0.0037). PD-L1 positivity status correlated with MMR-D by two algorithms. Five of 2268 (<1%) advansced CRCs demonstrated amplification of either the PD-L1 or PD-L2 genes by CGP. Conclusions PD-L1 expression in TC and IC is associated with advanced stage and MMR-D. PD-L1 positivity status by the published algorithm is associated with MMR-D. PD-L1 amplification is extremely uncommon in CRC. Evaluation of whole tissue section and incorporation of IC staining enhance the sensitivity to screen patients who may benefit from ICPI therapy.

Solid Tumors and HIV-Infected Patients Other Than AIDS-Defining Neoplasms

The first reported cases of Kaposi’s sarcoma (KS) in homosexual men appeared in 1981 and heralded the onset of the AIDS epidemic (1,2). A year later the first four cases of non-Hodgkin lymphoma (NHL) were reported (3). In 1983 the first cases of primary central nervous system lymphoma (PCL) were described (4). By 1985, NHL was added to KS and PCL as index AIDS-defining neoplasms (5). The incidence of PCL and NHL in particular is sharply on the rise (6,7). All three AIDS-defining neoplasms are characterized by higher grade lesions, more advanced stage, and shorter survival when compared to similar tumors in non-HIV infected patients. The epidemiology and clinical characteristics of these neoplasms is well described. (8–10). Today epidemic KS, PCL and NHL represent major causes of morbidity and mortality in AIDS patients.

Rectal ulcer and pseudomalignant epithelial changes after prostate seed brachytherapy: A rare complication with a diagnostic pitfall

BACKGROUND Implant brachytherapy (IBT) is a well-recognized treatment modality for early stage prostate cancer. Rectal ulcer and rectourethral fistula complicating IBT may cause an alteration of the normal anatomic landmarks. In this context, pseudomalignant radiation-induced changes within prostatic epithelium may be misinterpreted as a primary rectal malignancy. Such challenging and misleading findings have not been described, and may not be recognized as such. MATERIALS AND METHODS We present the clinical and pathologic aspects of two patients who underwent IBT for low stage prostate cancer that was complicated by deep rectal ulcer. Both patients underwent extensive palliative surgical resection for disease control. RESULTS The histologic changes in both cases were noteworthy for extensive necrosis and inflammation of the prostate, associated with loss of recto-prostatic anatomical landmarks. Prostatic glands showed striking radiation-induced atypia and pseudomalignant epithelial changes extending to the rectal ulcer bed, with no residual viable tumor. The first patient had undergone a biopsy of the rectal ulcer bed that was misinterpreted as a rectal adenocarcinoma prior to surgery. The similarity between atypical glands of the biopsy and the benign prostatic tissue with radiation-induced atypia in resection specimen confirmed their benign nature. CONCLUSIONS Deep rectal ulcer complicating IBT may lead to distortion of the normal recto-prostatic anatomical landmarks, resulting in detection of pseudo-malignant prostatic glands at the ulcer base. Such findings may be mistaken for a primary rectal malignancy in limited biopsy material if not familiar to the pathologist.

Abstract 4355: Prognostic significance of RUNX2 protein expression in carcinomas of the prostate (PAC), colon (CRC), and breast (BC)

Background: RUNX2, a member of the RUNX family of transcription factors, regulates normal bone, cartilage and skeletal muscle morphogenesis. Aberrant RUNX2 expression has correlated with malignant cell migration, invasion and bone metastases. In the following study, we evaluated the prognostic significance of RUNX2 protein expression in a large cohort of tumors from each of three sites of origin - prostate, colon and breast. Design: Formalin-fixed paraffin-embedded tissue sections from 121 PAC, 104 CRC and 82 invasive BC [64 ductal (IDC); 18 lobular (ILC)] were immunostained by a manual method (DAKO LSAB+ System-HRP) using mouse monoclonal RUNX2 (Santa Cruz). Nuclear and/or cytoplasmic immunoreactivity was scored based on intensity and percentage of positive cells in both the tumor (T) and adjacent benign (B) epithelium in each case. Cases were assessed as tumor>benign (T>B), tumor = benign (T = B), tumor Results: RUNX2 immunoreactivity was predominately nuclear in PAC, CRC and BC and noted as follows: in PACs: T>B 58%, T = B 31%, T = Gleason grade 7] vs low [ B 61%, T = B 22%, T B 65% and T = B 35%; and correlated overall with early [stage I, II] vs advanced [stage III, IV] tumor stage (p = 0.05); ER positive status (p = 0.028); and on Cox univariate analysis, correlated with both lengthened recurrence free (p = 0.033) and overall (p = 0.042) survival; and showed a trend toward association with PR positive status (p = 0.075). Within the ER negative BC subgroup, RUNX2 immunoreactivity correlated with early tumor stage (p = 0.008) and PR positive status (p = 0.033). Conclusion: While RUNX2 protein expression correlates with adverse prognostic variables in prostate and colon cancers, RUNX2 immunoreactivity portends favorable prognosis in breast cancer. Further study of RUNX2 expression and its potential role as a therapeutic target particularly in PAC and CRC appear warranted. Citation Format: Bhaskar V.S. Kallakury, Albert Huho, Ann B. Boguniewicz, David M. Jones, Tipu Nazeer, Hwa Jeong Lee, Christine E. Sheehan, Jeffrey S. Ross. Prognostic significance of RUNX2 protein expression in carcinomas of the prostate (PAC), colon (CRC), and breast (BC). [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4355. doi:10.1158/1538-7445.AM2015-4355