Celia Marginean

PIK3CA, BRAF and PTEN status and benefit from cetuximab in the treatment of advanced colorectal cancer - results from NCIC CTG / AGITG CO.17

Purpose: Cetuximab improves survival in patients with K-ras wild-type advanced colorectal cancer. We examined the predictive and prognostic significance of additional biomarkers in this setting, in particular BRAF, PIK3CA, and PTEN. Experimental Design: Available colorectal tumor samples were analyzed from the CO.17 study. BRAF mutations were identified in tumor-derived DNA by direct sequencing and PIK3CA mutations were identified using a high-resolution melting screen with confirmation by sequencing. PTEN expression by immunohistochemistry (IHC) was performed on tissue microarrays. For each biomarker, prognostic and predictive effects were examined using a Cox model with tests for treatment–biomarker interaction. Results: A total of 572 patients with pretreated colorectal cancer were randomly assigned to receive cetuximab or best supportive care (BSC). Of 401 patients assessed for BRAF status, 13 (3.2%) had mutations. Of 407 patients assessed for PIK3CA status, 61 (15%) had mutations. Of 205 patients assessed for PTEN, 148 (72%) were negative for IHC expression. None of BRAF, PIK3CA, or PTEN was prognostic for overall or progression-free survival in the BSC arm. None was predictive of benefit from cetuximab, either in the whole study population or the K-ras wild-type subset. In the K-ras wild-type subgroup, the overall survival adjusted HR according to BRAF mutation status was 1.39 (interaction P = 0.69), PIK3CA mutation status HR = 0.79 (interaction P = 0.63), and PTEN expression HR = 0.75 (interaction P = 0.61). Conclusions: In chemotherapy-refractory colorectal cancer, neither PIK3CA mutation status nor PTEN expression were prognostic, nor were they predictive of benefit from cetuximab. Evaluation of predictive significance of BRAF mutations requires a larger sample size. Clin Cancer Res; 20(3); 744–53. ©2013 AACR.

D2-40, a novel immunohistochemical marker in differentiating dermatofibroma from dermatofibrosarcoma protuberans

The distinction between dermatofibroma, particularly cellular variant, and dermatofibrosarcoma protuberans in excisional biopsies is usually straightforward. However, a separation between the two may be sometimes challenging, especially in superficial biopsies. Although factor XIIIa and CD34 immunostains are useful in differentiating dermatofibroma and dermatofibrosarcoma protuberans in most instances, focal CD34 positivity may be seen in cellular fibrous histiocytoma. Some cases reveal overlapping immunostain results. D2-40 identifies a 40-kDa O-linked sialoglycoprotein present on a variety of tissues including testicular germ cell tumors as well as lymphatic endothelium. In this study, we investigated the utility of D2-40 in separating dermatofibroma from dermatofibrosarcoma protuberans and compared the results with other commonly used immunostains. Fifty-six cases of dermatofibroma (including six cellular variant) and 29 cases of dermatofibrosarcoma protuberans were retrieved from the archives of Department of Anatomic Pathology at Sunnybrook Health Sciences Center in University of Toronto. We applied factor XIIIa, CD34, and monoclonal mouse anti-D2-40 immunostains to formalin-fixed, paraffin-embedded tissue sections. All 56 (100%) cases of dermatofibroma demonstrated strong and diffuse immunoreactivity to D2-40 in the spindle cells and stroma. Similarly, factor XIIIa showed strong and diffuse positivity in the spindle cells. Nearly all dermatofibromas were negative for CD34 except one case revealing focal positivity. None of dermatofibrosarcoma protuberans cases were labeled by D2-40, although four cases showed weak and patchy background staining in contrary to diffuse, strong, and crisp staining seen in dermatofibromas. Our results indicate that D2-40 seems to be a sensitive immunohistochemical marker for dermatofibromas, including cellular variant. Focal and faint D2-40 staining may be seen in the stroma of dermatofibrosarcoma protuberans. Our findings suggest that D2-40 can be used as a complementary immunostain to factor XIIIa and CD34 in problematic and challenging cases on superficial biopsies.

Diagnostic use of cytokeratins, CD34, and neuronal cell adhesion molecule staining in focal nodular hyperplasia and hepatic adenoma

Cytokeratins 7 and 19 and neuronal cell adhesion molecule (CD56) are differentially expressed in the hepatocytes and biliary epithelium. CD34 is an endothelial marker that is expressed in hepatic sinusoids in conditions associated with altered vascular flow and neoplasms. Distinct staining patterns using these markers have been shown in resected specimens of focal nodular hyperplasia, telangiectatic focal nodular hyperplasia, and hepatic adenoma. The purpose of this study was to examine the diagnostic use of these markers in needle biopsies. Needle biopsies from focal nodular hyperplasia (n = 21), telangiectatic focal nodular hyperplasia (n = 2), and hepatic adenoma (n = 14) were included in the study. These cases represent typical examples of each entity that have been diagnosed on the basis of clinical, imaging, and histologic features. Corresponding resection specimens available in 9 cases were also included in the study for comparison. Immunohistochemical analysis was performed on 4-mum-thick formalin-fixed and paraffin-embedded sections using antibodies against cytokeratin 7, cytokeratin 19, neuronal cell adhesion molecule, and CD34. The staining patterns and intensity for each marker were analyzed in a blinded fashion, and the patterns were recorded as focal nodular hyperplasia-like, hepatic adenoma-like, or indeterminate for each case. Presence of normal tissue was also recorded in each case. The hepatic adenoma-like pattern is characterized by strong cytokeratin 7 positivity in hepatocytes in patches with a gradual decrease in the staining intensity as the cells differentiate toward mature hepatocytes. Hepatic adenomas lack bile ducts and ductules as highlighted by cytokeratin 7, cytokeratin 19, and neuronal cell adhesion molecule stains. The focal nodular hyperplasia-like pattern is characterized by milder and focal cytokeratin 7 staining of hepatocytes. Cytokeratin 7, cytokeratin 19, and neuronal cell adhesion molecule show a strong staining of bile ductules in the fibrous septa. Normal liver shows cytokeratin 7, cytokeratin 19, and neuronal cell adhesion molecule staining of bile ducts, whereas the hepatocytes are generally negative. Of the 21 focal nodular hyperplasia cases, 20 cases (95.2%) showed a focal nodular hyperplasia-like pattern, whereas 13 (92.2%) of 14 hepatic adenoma cases showed a hepatic adenoma-like pattern. Both cases of telangiectatic focal nodular hyperplasia showed a hepatic adenoma-like pattern. CD34 stain showed areas of diffuse endothelial staining in 2 cases of hepatic adenoma, 3 cases of focal nodular hyperplasia, and both cases of telangiectatic focal nodular hyperplasia, whereas the remaining cases showed staining of endothelial cells only in the inflow areas of the sinusoids. A mixed (diffuse and inflow) pattern of CD34 staining was seen in 1 focal nodular hyperplasia, 1 hepatic adenoma, and 2 telangiectatic focal nodular hyperplasia cases. For statistical analysis, the telangiectatic focal nodular hyperplasia were considered as variants of hepatic adenoma. The findings were found to be highly statistically significant (P < .05) for cytokeratin 7, cytokeratin 19, and neuronal cell adhesion molecule stains. An inflow staining pattern favors a diagnosis of focal nodular hyperplasia; however, overall, CD34 stain was not helpful in differentiating focal nodular hyperplasia and hepatic adenoma. Corresponding resection specimens (hepatic adenoma = 5, focal nodular hyperplasia = 2) showed staining patterns that were identical to the biopsy, whereas resections of the telangiectatic focal nodular hyperplasia cases showed both focal nodular hyperplasia and hepatic adenoma-like areas. Considering that telangiectatic focal nodular hyperplasia is now thought to be a variant of hepatic adenoma, the staining patterns correctly identified all cases, except one case each of focal nodular hyperplasia and hepatic adenoma. In summary, a combination of cytokeratin 7, cytokeratin 19, and neuronal cell adhesion molecule immunostains performed on needle biopsies of liver shows distinctive patterns similar to that of resection specimen. The stains, especially cytokeratins 7 and 19, are very helpful in distinguishing normal from lesional tissue, as well as hepatic adenoma from focal nodular hyperplasia, and could be diagnostically helpful in challenging cases. Prospective studies to evaluate use of these stains in challenging cases are needed to validate these findings.

EGFR expression variance in paired colorectal cancer primary and metastatic tumors

Background: Previous studies indicate that drugs targeting the Epidermal Growth Factor Receptor (EGFR) signaling pathways can induce objective responses, prolong time to progression and improve survival of patients with metastatic colorectal cancer (mCRC). EGFR expression in the primary tumour may not predict response to these agents and data is conflicting regarding the correlation of EGFR expression in the primary tumour with the metastatic site. In other tumour sites, the presence of EGFR mutations was associated with efficacy in a subset of patients. Objectives: The goal of this study is to correlate tumour EGFR expression between primary and liver metastatic sites, and to assess the mutational status in the EGFR kinase domain. Methods: This is a single center retrospective study of patients who underwent surgical resection of CRC, for whom paired paraffin-embedded tissue blocks of primary tumours and resected liver metastases were available. EGFR immunostaining and mutation analyses were preformed. Results: Fifty eight paired colorectal primaries and metastases were available for analysis. EGFR was detectable in 96.6% of the primary samples and in 89.7% of the metastatic samples. Perfect concordance in the intensity score between the primary and the metastases was found in 46.5% of the cases. While individual pairs were poorly concordant for intensity, the proportion of primaries with intense staining was similar to the proportion with intense staining in the metastatic samples. Overall survival did not correlate with either EGFR expression in the primary tumour, or with EGFR expression in the metastasis. There were 2 cases with mutations in the EGFR kinase domain. Both mutations were found in exon21 C>T. Conclusions: In this analysis, EGFR expression in the primary tumor site was not predictive of its level in the metastasis. EGFR expression levels in the primaries and in the metastases do not appear to be useful prognostic markers.

The management of hepatobiliary cystadenomas: lessons learned

BACKGROUND Mucinous cystic neoplasms of the liver (hepatobiliary cystadenomas) are rare neoplastic lesions. Such cysts are often incorrectly diagnosed and managed, and carry a risk of malignancy. The objective of this study was to review the surgical experience with these lesions over 15 years. METHODS A retrospective chart review identified consecutive patients undergoing surgery for liver cystadenomas from 1997-2011. Clinical data were collected and summarized. RESULTS Thirteen patients (mean age 51 years, 12/13 females) with cysts 4.6-18.1 cm were identified. Most cysts were located in the left lobe/centrally (11/12) and had septations (8/13). Mural nodularity was infrequent (3/13). Nine patients had liver resection/enucleation, whereas four had unroofing. Frozen section analysis had a high false-negative rate (4/6). All patients had cystadenomas, of which two had foci of invasive carcinoma (cystadenocarcinoma) within mural nodules. There was no 90-day mortality. All but one patient (myocardial infarction) were alive at a median follow-up of 23.1 months. No patient with unroofing has developed malignancy to date. CONCLUSIONS Non-invasive hepatobiliary cystadenomas present as large central/left-sided cysts in young or middle-aged women. Associated malignancy was relatively uncommon and found within mural nodules. Intra-operative frozen section analysis was ineffective at ruling out cystadenomas. Complete excision is recommended, but close follow-up might be considered in patients with a prohibitive technical or medical risk, in the absence of nodularity on high-quality imaging.

Interobserver variability in assessing dysplasia and architecture in colorectal adenomas: a multicentre Canadian study

Aims Following the introduction of colorectal cancer screening programmes throughout Canada, it became necessary to standardise the diagnosis of colorectal adenomas. Canadian guidelines for standardised reporting of adenomas were developed in 2011. The aims of the present study were (a) to assess interobserver variability in the classification of dysplasia and architecture in adenomas and (b) to determine if interobserver variability could be improved by the adoption of criteria specified in the national guidelines. Methods An a priori power analysis was used to determine an adequate number of cases and participants. Twelve pathologists independently classified 40 whole-slide images of adenomas according to architecture and dysplasia grade. Following a wash-out period, participants were provided with the national guidelines and asked to reclassify the study set. Results At baseline, there was moderate interobserver agreement for architecture (K=0.4700; 95% CI 0.4427 to 0.4972) and dysplasia grade (K=0.5680; 95% CI 0.5299 to 0.6062). Following distribution of the guidelines, there was improved interobserver agreement in assessing architecture (K=0.5403; 95% CI 0.5133 to 0.5674)). For dysplasia grade, overall interobserver agreement remained moderate but decreased significantly (K=0.4833; 95% CI 0.4452 to 0.5215). Half of the cases contained high-grade dysplasia (HGD). Two pathologists diagnosed HGD in ≥75% of cases. Conclusions The improvement in interobserver agreement in classifying adenoma architecture suggests that national guidelines can be useful in disseminating knowledge, however, the variability in the diagnosis of HGD, even following guideline review suggests the need for ongoing knowledge-transfer exercises.

Gamut of focal fatty lesions in the liver: imaging manifestations with emphasis on magnetic resonance imaging.

A wide variety of hepatic lesions contain fat within them in a proportion good enough to be demonstrable on imaging. With the biological behaviors quite different from each other, they form a clinico-radio-pathologic spectrum worth remembering. We present a review of common and uncommon focal hepatic lesions containing fat (micro- and macroscopic) within them and emphasize the role of imaging in their diagnosis.

Oncocytic renal cell carcinoma with immunohistochemical properties of renal oncocytoma

Renal oncocytoma (RO) is a characteristic benign renal tumor. The existence of malignant RO is controversial and anecdotal, partly due to a lack of specific markers for RO. With recent advances in immunohistochemistry, RO can be distinguished from other renal neoplasms with routine stains and with the aid of immunostaining. We report two cases of renal neoplasms with similar histopathological appearances. They were characterized by oncocytic cytoplasm, numerous intra-cytoplasmic vacuoles, uniform round to oval hyperchromatic nuclei with remarkably thick nuclear membranes and prominent nucleoli. The tumor cells were closely packed and disposed in an alveolar pattern. The neoplastic cells were diffusely reactive for CD117 and progesterone receptor, and diffusely or focally reactive for cytokeratin AE1/AE3, and focally reactive for cytokeratin 7, CD10, and racemase. The cells were non-reactive for renal cell carcinoma (RCC) antigen, vimentin, S100, and neuroendocrine markers. One tumor showed lymph node metastasis. Due to the remarkable cytological atypia, lymph node metastasis, and similar immunological features of RO, these two tumors likely represent a distinct subtype of RCC related to RO.

Hepatic small vessel neoplasm, a rare infiltrative vascular neoplasm of uncertain malignant potential

Characteristic but rare vascular neoplasms in the adult liver composed of small vessels with an infiltrative border were collected from an international group of collaborators over a 5-year period (N=17). These tumors were termed hepatic small vessel neoplasm (HSVN), and the histologic differential diagnosis was angiosarcoma (AS). The average age of patients was 54years (range, 24-83years). HSVN was more common in men. The average size was 2.1cm (range, 0.2-5.5cm). Diagnosis was aided by immunohistochemical stains for vascular lineage (CD31, CD34, FLI-1), which were uniformly positive in HSVN. Immunohistochemical stains (p53, c-Myc, GLUT-1, and Ki-67) for possible malignant potential are suggestive of a benign/low-grade tumor. Capture-based next-generation sequencing (using an assay that targets the coding regions of more than 500 cancer genes) identified an activating hotspot GNAQ mutation in 2 of 3 (67%) tested samples, and one of these cases also had a hotspot mutation in PIK3CA. When compared with hepatic AS (n=10) and cavernous hemangioma (n=6), the Ki-67 proliferative index is the most helpful tool in excluding AS, which demonstrated a tumor cell proliferative index greater than 10% in all cases. Strong p53 and diffuse c-Myc staining was also significantly associated with AS but not with HSVN or cavernous hemangioma. There have been no cases with rupture/hemorrhage, disseminated intravascular coagulation, or Kasabach-Merritt syndrome. Thus far, there has been no metastasis or recurrence of HSVN, but complete resection and close clinical follow-up are recommended because the outcome remains unknown.

Eastern Canadian Colorectal Cancer Consensus Conference: standards of care for the treatment of patients with rectal, pancreatic, and gastrointestinal stromal tumours and pancreatic neuroendocrine tumours

The annual Eastern Canadian Colorectal Cancer Consensus Conference was held in Halifax, Nova Scotia, October 20-22, 2011. Health care professionals involved in the care of patients with colorectal cancer participated in presentation and discussion sessions for the purposes of developing the recommendations presented here. This consensus statement addresses current issues in the management of rectal cancer, including pathology reporting, neoadjuvant systemic and radiation therapy, surgical techniques, and palliative care of rectal cancer patients. Other topics discussed include multidisciplinary cancer conferences, treatment of gastrointestinal stromal tumours and pancreatic neuroendocrine tumours, the use of folfirinox in pancreatic cancer, and treatment of stage ii colon cancer.