Correlation of HER2 Protein Level With mRNA Level Quantified by RNAscope in Breast Cancer.

Trastuzumab deruxtecan (T-DXd) has been approved by the US Food and Drug Administration (FDA) to treat patients with metastatic HER2-positive and HER2-low breast cancer, and clinical trials are examining its efficacy against early-stage breast cancer. Current HER2 immunohistochemical (IHC) assays are suboptimal in evaluating HER2-low breast cancers and identifying which patients would benefit from T-DXd. HER2 expression in 526 breast cancer tissue microarray (TMA) cores was measured using the FDA-approved PATHWAY and HercepTest IHC assays, and the corresponding RNA levels were evaluated by RNAscope. HER2 protein levels by regression analysis using a quantitative immunofluorescence score against cell line arrays with known HER2 protein levels determined by mass spectrometry were available in 48 of the cores. RNAscope was also performed in 32 metastatic biopsies from 23 patients who were subsequently treated with T-DXd, and the results were correlated with response rate. HER2 RNA levels by RNAscope strongly correlated with HER2 protein levels (P < .0001) and with HER2 IHC H-scores from the PATHWAY and HercepTest assays (P < .0001). However, neither protein levels nor RNA levels significantly differed between cases scored 0, ultralow, and 1+ by PATHWAY and HercepTest. The RNA levels were significantly higher (P = .030) in responders (6.4 ± 8.2 dots/cell, n = 12) than those in nonresponders (2.6 ± 2.2, n = 20) to T-DXd. RNAscope is a simple assay that can be objectively quantified and is a promising alternative to current IHC assays in evaluating HER2 expression in breast cancers, especially HER2-low cases, and may identify patients who would benefit from T-DXd.

Automated RNA In Situ Hybridization for 18 High Risk Human Papilloma Viruses in Squamous Cell Carcinoma of the Head and Neck: Comparison With p16 Immunohistochemistry.

Detection of human papilloma virus (HPV)-related head and neck squamous cell carcinoma (HNSCC) is important, as HPV-associated HNSCCs respond better to therapy. The RNAscope HPV-test is a novel RNA in situ hybridization (ISH) technique which strongly stains transcripts of E6 and E7 mRNA in formalin-fixed, paraffin-embedded tissue, with the potential to replace the indirect immunohistochemical (IHC) marker for p16 protein. A direct clinical comparison between p16 IHC and an automated RNA ISH using 18 probes has not been established. Samples from 27 formalin-fixed, paraffin-embedded HNSCC cases from the Emory University Hospital archives were stained using 18 individual RNA ISH probes for high-risk HPV (RNAscope 2.5 LS Probe ) on a Leica autostainer (Buffalo Grove, IL) and were compared with p16 IHC. Two pathologists reviewed and reached a consensus on all interpretations. The RNAscope technique was positive in 89% (24/27) and the p16 IHC was positive in 78% (21/27). The RNAscope was negative in 11.1% of samples (3/27) and the p16 IHC-negative in 22.2% (6/27). The RNA ISH detected 100% of the p16-positive IHC-stained slides and had a concordance of 88.9% (24/27). This easy to interpret automated staining method for 18 high-risk HPV genotypes is a feasible replacement for the indirect p16 IHC method.

RNA In Situ Hybridization for Epstein-Barr Virus and Cytomegalovirus: Comparison With In Situ Hybridization and Immunohistochemistry.

The RNAscope utilizes in situ hybridization (RISH) technology to detect single RNA molecules in a variety of tissue samples, including formalin fixed paraffin embedded (FFPE) tissues. Epstein-Barr virus (EBV) and cytomegalovirus (CMV) are found in association with neoplastic tissues and inflammatory lesions, and immunohistochemistry (IHC) or other techniques (ISH) are utilized to identify them. We compared the RNAscope RISH to ISH and IHC in the detection of EBV and CMV respectively to determine RNAscope utility in a clinical setting. Thirty-one FFPE tissues were stained by RISH to detect EBV and 24 samples of tissue for CMV. The RISH used the RNAscope (Leica BioSystems, Buffalo Grove, IL), the Bond III autostainer (Leica), and probes V-EBV and V-CMV (Advanced Cell Diagnostics, Newark, CA) as well as negative (DapB) and positive probe (PPIB) for RNA. Results were compared with those by ISH (Leica, EBV RNA probe), and IHC (CMV Dako, 1/160), respectively. RISH and ISH were concordant in 100% of cases positive for EBV by ISH (19/19). Of the cases negative for EBV by ISH, RISH showed positivity in an additional 25% of the samples (3/12). Overall concordance was 90.3% (28/31). RISH and IHC were concordant in 100% of cases positive for CMV by IHC (8/8). Of the cases negative for CMV by IHC, RISH detected positivity in an additional 50% of the samples (8/16). Overall concordance was 66.7% (16/24). RISH demonstrates increased sensitivity in the clinical setting, especially for CMV, detecting positive cells not stained by EBV ISH and CMV IHC.

Renal Medullary and Cortical Correlates in Fibrosis, Epithelial Mass, Microvascularity, and Microanatomy Using Whole Slide Image Analysis Morphometry.

Renal tubulointerstitial injury often leads to interstitial fibrosis and tubular atrophy (IF/TA). IF/TA is typically assessed in the renal cortex and can be objectively quantitated with computerized image analysis (IA). However, the human medulla accounts for a substantial proportion of the nephron; therefore, medullary scarring will have important cortical consequences and may parallel overall chronic renal injury. Trichrome, periodic acid-Schiff (PAS), and collagen III immunohistochemistry (IHC) were visually examined and quantitated on scanned whole slide images (WSIs) (N = 67 cases). When tuned to measure fibrosis, IA of trichrome and Trichrome-PAS (T-P) WSIs correlated for all anatomic compartments (among cortex, medulla, and entire tissue, r = 0.84 to 0.89, P all <0.0001); and collagen III deposition correlated between compartments (r = 0.69 to 0.89, P <0.0001 to 0.0002); however, trichrome and T-P measures did not correlate with collagen deposition, suggesting heterogeneous contributions to extracellular matrix deposition. Epithelial cell mass (EPCM) correlated between cortex and medulla when measured with cytokeratin IHC and with the trichrome red portion (r = 0.85 and 0.66, respectively, all P < 0.0001). Visual assessment also correlated between compartments for fibrosis and EPCM. Correlations were found between increasing medullary inner stripe (IS) width and fibrosis in all of the tissue and the medulla by trichrome morphometry (r = 0.56, P < 0.0001, and r = 0.48, P = 0.00008, respectively). Weak correlations were found between increasing IS width and decreasing visual assessment of all tissue EPCM. Microvessel density (MVD) and microvessel area (MVA) measured using a MVD algorithm applied to CD34 IHC correlated significantly between all compartments (r = 0.76 to 0.87 for MVD and 0.71 to 0.87 for MVA, P all < 0.0001). Overall, these findings demonstrate the interrelatedness of the cortex and medulla and the importance of considering the renal parenchyma as a whole.

Automated in situ hybridization for human papilloma virus.

With the increased number of requests for high-risk human papilloma virus in situ hybridization (HPV ISH), not only for uterine cervical squamous cell carcinoma (SCC) but also for biopsies and resections of oropharyngeal SCC, and fine needle aspiration cell blocks of metastatic SCC in cervical lymph nodes, we optimized an automated method to replace the manual one we had used. The new technique used the Leica BOND-maX or III immunostainer, already in use for immunohistochemical analysis, with the Enzo HPV type 16/18 and 6/11 probes. We stained 55 surgical biopsies and 41 cell blocks of oropharyngeal SCC. Sensitivity was 90% and 73% for biopsies and cell blocks, respectively, with a specificity of 100% in both. Stain is strong and crisp with no background. Turnaround time is short as runs are performed daily, with up to 30 slides per run. Technologists become trained in a few days, and the repeat rate is low. The only disadvantage is that ISH and IHC cannot be performed simultaneously. We highly recommend this automated technique for high-risk and low-risk HPV ISH and probably with other probes.

Galectin-3 and PTEN expression in pancreatic ductal adenocarcinoma, pancreatic neuroendocrine neoplasms and gastrointestinal tumors on fine-needle aspiration cytology.

OBJECTIVE: Galectin-3 has been implicated in the carcinogenesis of pancreatic ductal adenocarcinoma (PDAC). Its applicability in pancreatic fine-needle aspiration (FNA) in separating malignant from benign lesions has never been addressed. In addition, a correlation between Galectin-3 and tumor suppressor phosphatase and tensin homolog (PTEN) and their potential diagnostic value has never been tested. STUDY DESIGN: This study analyzed Galectin-3 immunohistochemical expression in FNA cell blocks of PDAC, pancreatic neuroendocrine neoplasms (PNEN), gastrointestinal stromal tumors (GIST) and non-tumor pancreatic tissue. In parallel, Galectin-3 and PTEN levels were evaluated in a tumor tissue microarray (TMA). RESULTS: Forty-four of 46 PDAC FNA and 32 of 33 PDAC TMA demonstrated tumor-specific Galectin-3 positivity. In contrast, Galectin-3 was not detected in PNEN and GIST. Total loss of PTEN was displayed by 26 of 33 PDAC, while non-neoplastic tissues all retained PTEN expression. CONCLUSION: Galectin-3 could be a valuable marker to help diagnose PDAC and rule out PNEN and GIST. In addition, PTEN positivity strongly argues against a diagnosis of PDAC. These data also advocate their potential diagnostic roles in the work up of challenging cytologic cases requiring ancillary test confirmation.

Tumor stem cells (CD271, c-kit, SOX10) in Melanomas: prognostic and outcome implications.

Melanoma cells that express stem cell marker CD271 are shown to form tumors when transplanted into nude or immunodeficient mice. These tumors have a higher metastatic potential and worse prognosis than melanomas resulting from transplantation of CD271-negative cells. We studied stem cell markers (CD271, c-kit, SOX1O) in melanomas, correlating their presence with prognostic factors and outcome. A total of 82 melanomas in tissue microarrays were immunostained for CD271, c-kit, and SOX10. Results were correlated with clinicopathologic prognostic parameters (Breslow depth of invasion, Clark level, sentinel lymph node status, and pathologic stage) and outcome (recurrence, metastases, and death). Of the 82 melanomas, CD271 was expressed in 18 (21%), c-kit in 47 (57%), and SOX10 in all (100%). CD271 does show correlation with metastases (P=0.05). c-kit is associated with favorable prognostic parameters [Breslow depth (P<0.001) and pathologic stage (P=0.02)] and with improved outcome [recurrence (P=0.03) and metastases (P=0.004)]. Although SOX10 is a good diagnostic marker, it cannot be used for prognosis because it is expressed in all the melanomas studied. In conclusion, CD271 expression in melanomas is associated with increased frequency of metastases, and c-kit immunoreactivity is associated with favorable prognostic parameters and improved outcome.

Automated and manual human papilloma virus in situ hybridization and p16 immunohistochemistry: comparison in metastatic oropharyngeal carcinoma.

OBJECTIVES: We compared the efficacy of automated and manual human papilloma virus (HPV) in situ hybridization (ISH) and p16 immunohistochemical staining (IHC) in fine needle aspiration (FNA) of metastatic oropharyngeal carcinoma. STUDY DESIGN: A total of 41 FNA cell blocks (CB) were evaluated. HPV ISH was interpreted as positive if a minimum of one tumor cell showed punctate dot-like nuclear positivity. p16 was interpreted as positive if ≥70% of tumor cells showed brown nuclear and cytoplasmic staining. RESULTS: Thirty of 41 CB (73%) were positive by automated HPV ISH, 25 of 41 CB (60%) with manual HPV ISH. Eighteen of 41 CB (43%) were positive for p16 IHC. Twelve of 41 CB (29%) with automated HPV ISH and 2 of 41 CB (4%) with the manual method were positive at 10× magnification. Three of 41 CB (7%) with automated HPV ISH and 14 of 41 CB (34%) with the manual method were positive at 20× magnification. Fifteen of 41 CB (36%) with automated HPV ISH and 9 of 41 CB (21%) with the manual method were positive at 40-60× magnification. CONCLUSION: Automated HPV ISH plays a more significant role in determining the HPV status in CB. However, the failure to use high magnification in the evaluation can give false-negative results.

Phosphohistone H3 and Ki-67 labeling indices in cytologic specimens from well-differentiated neuroendocrine tumors of the gastrointestinal tract and pancreas: a comparative analysis using automated image cytometry.

BACKGROUND: Ki-67 proliferation index was recently incorporated in the grading of neuroendocrine neoplasms (NENs) of the gastrointestinal tract (GIT) and pancreas. These are now divided into well-differentiated neuroendocrine tumors (WDNETs, grades 1 and 2) and poorly differentiated neuroendocrine carcinomas (grade 3). While Ki-67 is an established proliferation marker in NENs, phosphohistone H3 (PHH3), a newer marker of mitotic activity, is not. METHODS: We determined Ki-67 and PHH3 indices on cytologic samples from WDNETs of the GIT and pancreas using an automated cellular imaging system (ACIS®). RESULTS: There was a strong correlation between Ki-67 and PHH3 indices generated by ACIS on cytologic samples. However, in some cases the two stains caused conflicting grades within the same tumor. CONCLUSION: Both antibodies stain cells in different phases of the cell cycle which may cause discordant grades, thus affecting patient management and prognostication. Ki-67 staining is stronger than PHH3, making 'hot spots' easier to identify on ACIS. Ki-67 is more ideal than PHH3 for staining NENs, especially in tumors with borderline grades. Because PHH3 generates lower mitotic indices it should not be used as a proliferation marker in NENs until its expression has been further characterized.

Image cytometric HER2 in gastric carcinoma: is a new algorithm needed?

BACKGROUND: Algorithms for quantitation of HER2 immunohistochemistry were developed for breast carcinoma, where the membranous stain must be entirely around the cell membrane. For gastric carcinoma, although assessment of intensity of immunostain (0 to 3) is similar, the site and percentage of stain differs by lacking the requirement of entire cell membrane positivity (complete, basolateral, or lateral membranous reactivity is sufficient for a positive result). We quantitated HER2 in gastric cancer specimens visually and by image cytometry, comparing results, and where available, with fluorescence in situ hybridization (FISH). The goal was to assess whether lack of concordance among results, might suggest a requirement for changing the image cytometric algorithm. DESIGN: All gastric carcinoma biopsies, resections, and cell blocks studied for HER2 expression/amplification in the past 2 years were included. Immunostain intensity, percentage, and score 0 to 3+ (0, 1+ negative, 2+ equivocal, 3+ positive), were evaluated visually, and by image cytometry with the ACIS score 0 to 3 (0, 1 negative, 2 equivocal, 3 positive). FISH (<1.8 negative, 1.8 to 2.2 equivocal, >2.2 amplified) was performed on all specimens with scores 2 and 3 by image cytometry. Results were compared. RESULTS: Sixty-eight specimens were studied, including 43 (63.2%) biopsies, 17 (25%) resections, and 8 (11.8%) cell blocks. Forty-seven (69.1%) were primary gastric, esophageal, or gastroesophageal junction adenocarcinoma; 19 (27.9%) were metastatic; 3 (4.4%) were well, 14 (20.6%) moderately (17, 25% tubular), and 51 (75%) poorly differentiated (poorly cohesive). Fourteen (20.6%) of cases were HER2 IHC positive with no significant difference in frequency based on type of specimen, site of carcinoma, or differentiation. Of the 14 visually HER2 IHC positive, 13 were positive by image cytometry (93% concordance), all 13 were amplified by HER2 FISH (100% concordance). Of the 3 cases equivocal both visually and by image cytometry, only 1 was FISH amplified. Fifty-one were negative by IHC visually and 52 by image cytometry (98% concordance). None of the 5 HER2 IHC negative were amplified by FISH. CONCLUSIONS: Despite different recommendations for interpretation of HER2 in gastric versus breast cancer, equivocal and positive/amplified results visually, and by image cytometry, and where FISH was performed, are similar. This concordance is noted for biopsy, resection, and cell block specimens, for primary versus metastatic, and for moderately versus poorly differentiated carcinoma; HER2 positivity/amplification is most frequent with poor differentiation, but not significantly so. There seems to be no need for the HER2 image cytometric algorithm used for breast cancer, to be changed when used for assessment of gastric cancers.

SOX10 expression in malignant melanoma, carcinoma, and normal tissues.

Sry-related HMg-Box gene 10 (SOX10) is a nuclear transcription factor that plays an important role in melanocytic cell differentiation. It has been shown to be a sensitive marker of melanoma including spindle and desmoplastic subtypes. We assessed its frequency of expression in melanoma, carcinoma, benign nevi, and non-neoplastic tissues with routine immunohistochemistry for SOX10. The 109 primary melanoma included 49 epithelioid, 19 spindle cell, 22 desmoplastic, and 19 mixed spindle cell/desmoplastic melanoma. All primary, except 8 desmoplastic melanoma, and 11 metastatic melanoma were strongly and diffusely nuclear SOX10-positive. Six desmoplastic melanoma had ≤10% cells positive, and 2 were <50% positive, all of 3+ intensity. Eighteen of 149 (12%) breast carcinoma were SOX10-positive. All 24 ovarian, 23 endometrial, 26 lung, and 25 colon carcinoma were SOX10-negative. All 43 benign nevi, 18 dysplastic nevi, 68 non-neoplastic and benign skins, and all 56 non-neoplastic breast tissue were SOX10-positive. The sensitivity and specificity for SOX10 in the diagnosis of melanoma are 1.0 and 0.93, respectively; the positive and negative predictive values are 0.87 and 1.0, respectively. SOX10 is a sensitive, specific marker for melanoma. As benign nevi also express SOX10, it cannot be used to differentiate between benign and malignant pigmented skin lesions. Only a small number of breast carcinoma (12%), and breast lobules, express SOX10; no carcinoma of the ovary, endometrium, lung, or colon expressed SOX10.

Inference of the Basal epithelial phenotype in breast carcinoma from differential marker expression, using tissue microarrays in triple negative breast cancer and women younger than 35.

Basal-cell phenotype breast carcinoma has been associated with high-grade and metaplastic morphology, expression of basal-type cytokeratins, uniform negativity for ER and HER2, and decreased overall survival. Breast cancers occurring in young women are usually T2 disease at presentation, high-grade and of poor prognosis. We compared two groups of breast cancers, (a) ER-, PR-, HER2- (triple negative) [TNBrCa] and (b) non-triple negative breast cancers (non-TNBrCa) occurring in women under 35, using tissue microarray technology to characterize expression of the basal/myoepithelial cytokeratins (CK5/6, CK7, and CK14), luminal cytokeratins (CK8, CK18, and CK19), EGFR, p-cadherin, c-kit, p63, and p53. We also sought to identify characteristic histomorphologic features indicative of basal-like phenotype. The triple negative group showed preferential staining versus the age <35 group for CK5/6 (22% versus 4% p = 0.05), CK14 (44% versus 15%, p = 0.013), EGFR (83% versus 24%, p < 0.0001) and c-kit (19% versus 0% p = 0.026). Conversely, non-TNBrCa in women younger than 35 demonstrated increased expression of the luminal CK8 (92% versus 60%) compared with the triple negative patients (p = 0.006). The TNBrCa have characteristic histologic features including higher tumor grade, pushing tumor border, geographic necrosis, syncytial growth pattern, brisk mitotic activity, lack of/minimal in situ component, medullary-like and metaplastic differentiation. Invasive carcinomas in women younger than 35 usually have an associated in situ component, prominent nucleoli, central acellular fibrotic zone, and infiltrative tumor border. Triple negativity for ER/PR/HER2 coupled with EGFR, c-kit, and basal/myoepithelial cytokeratins (CK5/6, CK14) expression, and distinctive histomorphologic features predict morphology consistent with basal-cell phenotype.

Combined double CK5/P63 stain: useful adjunct test for diagnosing pulmonary squamous cell carcinoma.

Increasing demand for accurate differentiation of pulmonary squamous cell carcinoma (SQCC) from other subtypes can be challenging for pathologists. This is more so in fine-needle aspirations (FNA) since the sample is small and SQCC may show degenerative changes and necrosis that distort the cellular features. Immunohistochemistry (IHC) is a valuable adjunct, and CK5/6 and P63 immunoreactivity is found to be basically restricted to SQCC. In our study, we evaluated the efficiency of CK5/P63 double staining in the diagnosis of pulmonary SQCC in cell blocks (CB) of lung FNA. We used a cohort including 24 CB of lung SQCC and 34 CB of lung adenocarcinomas (ADC). IHC was performed for CK5/P63 double stain. Seventeen of 24 (70%) lung SQCC were positive for the double stain CK5/P63. Two (8%) were positive for CK5 alone and two (8%) were positive for P63 alone. Thus, a total 19 of 24(79%) SQCC of the lung were positive for CK5 and P63 each. In ADC, no immunoreactivity was detected for CK5 alone or combined CK5/P63. Three of 34(8%) ADC were positive for P63. This first study of double staining of CK5/P63 in FNA CB shows a sensitivity of 70% and specificity of 100% for SQCC of the lung. When each marker staining alone is included, the sensitivity for CK5 and P63 increases to 79% each. This double stain can help in the diagnosis of pulmonary SQCC with an accuracy of 88% and a positive predictive value of 100%.

TTF-1 and Napsin A double stain: a useful marker for diagnosing lung adenocarcinoma on fine-needle aspiration cell blocks.

BACKGROUND: Immunohistochemistry (IHC) for thyroid transcription factor-1 (TTF-1) is used to confirm the diagnosis of lung adenocarcinoma. Napsin A also has shown positivity in lung adenocarcinoma. A combined double stain for TTF-1 and napsin A has been proposed to achieve higher sensitivity and specificity. In this study, the authors evaluated the utility of this double stain in the diagnosis of lung adenocarcinoma in cell blocks of fine-needle aspirates (FNA). METHODS: The authors used a cohort comprising 35 FNA cell blocks of lung adenocarcinoma and 24 FNA cell blocks of lung squamous cell carcinoma (SqCCA). IHC was performed; expressions of TTF-1 as brown nuclear stain and of napsin A as red cytoplasmic stain were identified. RESULTS: Twenty-six of 35 (74%) lung adenocarcinomas were positive for double staining with TTF-1/napsin A. Of 35 lung adenocarcinomas, only 2 (5%) were positive for TTF-1 alone and 3 (8%) were positive for napsin A alone. For the double stain TTF-1/napsin A, 3 of 24 (12%) lung SqCCAs were positive for both. Six of 24 (25%) cases were positive for TTF-1 alone, and none were positive for napsin A alone. For lung adenocarcinoma, TTF-1/napsin A has a sensitivity of 74%, specificity of 87%, accuracy of 79%, and a positive predictive value of 89%. CONCLUSIONS: The double IHC stain, TTF-1/napsin A, for the identification of pulmonary adenocarcinoma in FNA cell block materials was diagnostically useful. The use of napsin A alone demonstrated a greater degree of accuracy and appeared diagnostically useful as a single IHC stain.

Image cytometric validation of breast carcinoma markers (ER, HER2 and MIB-1) using tissue microarrays: rabbit monoclonal vs. FDA-approved antibodies.

OBJECTIVE: To use the ACIS III (Dako, Carpinteria, California, U.S.A.) with tissue microarrays (TMAs) to compare rabbit monoclonal antibodies (RMab) for ER, HER2, and MIB-1 with FDA-approved monoclonal (FMab) and polyclonal (FPab) antibodies. STUDY DESIGN: TMAs of 43 breast cancers were used. Immunohistochemistry was performed using RMab (LabVision, Fremont, California, U.S.A.): ER (SP1; 1/100), HER2 (SP3; 1/100), and MIB-1 (SP6; 1/200). FMPab (Dako) used: ER (1D5; 1/50), HercepTest kit and MIB-1 (MIB-1; 1/160). The stained TMAs were quantitated visually and by image cytometry (ACIS III). RESULTS: The overall agreement between RMab and FMab for ER using visual (98.45%) and image analysis (97.56%) was excellent, with a kappa level of 0.89 and 0.94, respectively. For HER2, the overall agreement between RMab and FPab was fair for visual (67.44%) and substantial (87.50%) for image analysis, with a kappa level of 0.32 and 0.72, respectively. For MIB-1, there was fair (64.29%) to poor (43.33%) agreement between MIB-1 RMab and FMab using visual and image analysis, with a kappa level of 0.47 and 0.16, respectively. CONCLUSION: RMabs for ER (SP1) and HER2 (SP3) are almost comparable to their counterpart, FDA antibodies; however, MIB-1 RMab (SP6) shows poor concordance with FMab in TMA. Image analysis shows a better concordance than visual quantitation assessment specifically for ER and HER2.

Collaboration to partnerships.

Partnerships are at the center of the Hospital of the University of Pennsylvania Nursing Excellence Professional Practice (HUP-NEPP) model. Through the use of collaboration, skilled communication, and respectful workplace, partnerships can be formed, leading ultimately to world-class patient care. At HUP, interdisciplinary partnerships are evidenced by the clinical nurses through shared governance. This article describes the components necessary to form successful partnerships.

Triple-negative breast carcinoma in women from Vietnam and the United States: characterization of differential marker expression by tissue microarray.

Triple-negative breast carcinoma accounts for approximately 15% of all breast cancers. It is characterized by an aggressive clinical history, high rate of local relapse, and association with the basal epithelial-like subtype. Variations in breast cancer subtype and clinical outcome often exist across racial and ethnic lines. Therefore, the aim of this study was to compare the immunohistochemical and clinicopathologic characteristics of triple-negative breast carcinoma in women living in Vietnam with those from the United States. Invasive triple-negative breast carcinoma of patients from the 2 populations was characterized by tissue microarray for the expression of basal cytokeratins (CK5/6, CK7, CK14), luminal cytokeratins (CK8, CK18, CK19), and markers associated with the basal phenotype (cKit, epithelial growth factor receptor, P-cadherin, p53, and p63). Significant differences in expression between the 2 populations were not observed for the basal cytokeratins. However, epithelial growth factor receptor and P-cadherin, markers associated with the basal phenotype, were underexpressed in Vietnamese patients. Of the luminal cytokeratins, CK8 was overexpressed and CK18 was underexpressed in the Vietnamese women. Significant differences were also observed regarding the clinicopathologic characteristics. Triple-negative breast carcinoma in Vietnamese women was smaller and less likely to be grade III. In addition, it was more frequently of ductal histologic type and less often medullary or metaplastic. These differences in histology and marker expression suggest that triple-negative breast carcinoma has unique biological characteristics in women from Vietnam and the United States, and may follow a unique clinical course in each of the 2 populations.

Claudin-7 and claudin-8: immunohistochemical markers for the differential diagnosis of chromophobe renal cell carcinoma and renal oncocytoma.

Claudin-7 and claudin-8 code for tight junction proteins expressed in distal nephron epithelium. In a recent oligonucleotide microarray study, we identified claudin-7 and claudin-8 as candidate markers to distinguish chromophobe renal cell carcinoma from other renal tumors, including oncocytoma. Distinction of these lesions can be difficult by light microscopy but is clinically important because chromophobe renal cell carcinoma has malignant biological potential, whereas renal oncocytoma is benign. Claudin-7 and claudin-8 expression was studied by immunohistochemistry in 11 chromophobe renal cell carcinomas and 17 oncocytomas using formalin-fixed paraffin-embedded tissue sections of tumor with adjacent nonneoplastic kidney. Steam antigen retrieval was performed before immunohistochemistry. Specificity was verified by negative control reactions without primary antibody and appropriate membranous staining patterns in positive control tissues (colon carcinoma and adjacent nonneoplastic kidney). Claudin-7 protein was expressed in a membranous pattern in 10 of 11 chromophobe renal cell carcinomas and 4 of 17 oncocytomas (P < .01). Claudin-8 was expressed in multiple patterns: In oncocytoma, 11 of 17 cases showed cytoplasmic, 4 of 17 membranous, and 2 of 17 negative reactions. In chromophobe renal cell carcinoma, 0 of 11 cases showed cytoplasmic, 3 of 11 membranous, and 8 of 11 negative reactions (P < .01). The immunohistochemical pattern of membranous claudin-7 and negative claudin-8 was seen in 7 of 11 chromophobe renal cell carcinomas and 1 of 17 oncocytomas (63% sensitivity, 84% specificity, 88% positive predictive value for chromophobe renal cell carcinoma). Negative claudin-7 and cytoplasmic claudin-8 were observed in 10 of 17 oncocytomas and 0 of 11 chromophobe renal cell carcinomas (59% sensitivity, 100% specificity and positive predictive value for oncocytoma). The distal nephron proteins claudin-7 and claudin-8 have potential use as immunohistochemical biomarkers in the differential diagnosis of chromophobe renal cell carcinoma and oncocytoma. Expression of claudin-7 and claudin-8 may reflect the relationship of chromophobe renal cell carcinoma and oncocytoma to intercalated cells of the cortical collecting duct. It may be necessary to identify additional biomarkers to include with claudin-7 and claudin-8 in a larger immunohistochemical panel to improve diagnostic sensitivity and specificity.

GATA-4 and GATA-6 expression in human ovarian surface epithelial carcinoma.

GATA-4 and GATA-6 are zinc finger transcription factors named for their recognition motif and involved in ovarian development and function. GATA factors are strongly expressed and primarily localized within the nuclei of ovarian surface epithelial cells. GATA factors have been previously shown to be expressed in sex-cord stromal ovarian tumors and may contribute to the tumor phenotype. Differential expression of GATA-4 within serous and mucinous ovarian carcinomas has been reported. Using immunohistochemistry, we studied GATA-4 and GATA-6 expression in 50 ovarian surface epithelial carcinomas and examined the relationship to clinicopathologic parameters and outcome. We found that the majority of the carcinomas retained GATA-4 expression, whereas approximately two-thirds of the carcinomas had mislocalization or loss of GATA-6 expression. No statistically significant correlations were found between histologic type, histologic grade, or patient outcome and GATA-4. Cytoplasmic GATA-6 expression tended to correlate with overall survival (P=0.0756). These findings suggest that although GATA factors play a role in ovarian surface epithelial carcinoma oncogenesis, they do not seem to affect clinicopathologic parameters.

Survivin and caspase-3 expression in breast cancer: correlation with prognostic parameters, proliferation, angiogenesis, and outcome.

BACKGROUND: Survivin is an inhibitor of apoptosis protein that is overexpressed in most human cancers, including breast, but is not expressed in normal tissue. Survivin is associated with more aggressive behavior and decreased survival in a variety of tumor types. It regulates the G2/M phase of the cell cycle by associating with mitotic spindle microtubules, and it directly inhibits caspase-3 and caspase-7 activity. We used a breast cancer tissue microarray to assess survivin and caspase-3 expression in breast cancer and to correlate both markers with proliferation (MIB-1), angiogenesis (CD31), and prognosis. DESIGN: A breast cancer tissue microarray with a total of 190 1-mm tissue samples (2 from each specimen) were immunostained for survivin, caspase-3, MIB-1, and CD31. The microarray contains 91 cases of breast carcinoma diagnosed at Emory University Hospital between 1992 and 2000, and 4 normal breast tissue controls. Follow-up information was obtained from hospital records and the Winship Cancer Center database. RESULTS: Eighty-four percent of breast carcinoma showed nuclear survivin expression. Normal breast tissue was immunonegative. Fifty-seven percent and 43% of breast cancer showed reduced and absent caspase-3 expression, respectively. Survivin (nuclear) and caspase (nuclear/cytoplasmic) expression showed significant correlation with histologic grade (P=0.008 and 0.041) and MIB-1 expression (P=0.033 and 0.012). Survivin nuclear expression also correlated significantly with tumor stage (P=0.012) and tended to correlate with estrogen receptor (P=0.050). There was no significant correlation between survivin and caspase expression. Furthermore, there was no correlation of both markers with other clinicopathologic parameters (age, tumor size, histologic type, progesterone receptor, Her-2 neu status, lymph node status), angiogenesis (CD31), or outcome (overall and disease-free survival). CONCLUSIONS: Survivin and caspase-3 expression correlate with poor prognostic parameters (higher histologic grade and high proliferation), but not with outcome, in breast carcinoma patients.