Amanda Jagdis

Prospective Evaluation of Tle1 as a Diagnostic Immunohistochemical Marker in Synovial Sarcoma

Synovial sarcoma is a high-grade soft tissue sarcoma that can be challenging to diagnose on the basis of histology alone. It is defined by a characteristic translocation t(X;18) that produces the fusion oncogene SYT-SSX. The current diagnostic gold standard for synovial sarcoma is the demonstration of the translocation by fluorescence in situ hybridization, reverse transcriptase polymerase chain reaction, or cytogenetics, in an appropriate histologic context. TLE1 encodes a transcriptional corepressor that is overexpressed in synovial sarcomas. Gene and tissue microarray studies have identified TLE1 as an excellent bio-marker for distinguishing the synovial sarcoma from other soft tissue malignancies. We prospectively evaluated incoming soft tissue tumor cases where the histology and clinical setting made synovial sarcoma a real consideration in the differential diagnosis. TLE1, Bcl2, epithelial membrane antigen, and cytokeratin expression were assessed using commercially available antibodies. TLE1 gave intense, diffuse nuclear staining in 35 of 35 molecularly confirmed synovial sarcoma cases, and was rare to absent in the 73 other soft tissue tumors examined (positive staining was found only in 1 of 43 malignant peripheral nerve sheath tumors, the 1 tested fibrosarcoma, and 1 pleomorphic sarcoma). TLE1 was more sensitive and specific for synovial sarcoma than other currently available immunohistochemical markers including Bcl2, epithelial membrane antigen and cytokeratins, and had a positive predictive value of 92% and a negative predictive value of 100% in this clinical setting. Our findings confirm, in a prospective diagnostic context, that TLE1 is more sensitive and specific for synovial sarcoma than any other currently available immunohistochemical stains, and in some cases may preclude the need for molecular testing.

Platelets in the immune response: Revisiting platelet-activating factor in anaphylaxis

Anaphylaxis is an acute, severe, life-threatening multisystem allergic reaction resulting from the sudden systemic release of biochemical mediators and chemotactic substances. Release of both preformed granule-associated mediators and newly generated lipid-derived mediators contributes to the amplification and prolongation of anaphylaxis. Platelet-activating factor (PAF) is a potent phospholipid-derived mediator the central role of which has been well established in experimental models of both immune-mediated and non-immune mediated anaphylaxis. It is produced and secreted by several types of cells, including mast cells, monocytes, tissue macrophages, platelets, eosinophils, endothelial cells, and neutrophils. PAF is implicated in platelet aggregation and activation through release of vasoactive amines in the inflammatory response, resulting in increased vascular permeability, circulatory collapse, decreased cardiac output, and various other biological effects. PAF is rapidly hydrolyzed and degraded to an inactive metabolite, lysoPAF, by the enzyme PAF acetylhydrolase, the activity of which has shown to correlate inversely with PAF levels and predispose to severe anaphylaxis. In addition to its role in anaphylaxis, PAF has also been implicated as a mediator in both allergic and nonallergic inflammatory diseases, including allergic rhinitis, sepsis, atherosclerotic disease, and malignancy, in which PAF signaling has an established role. The therapeutic role of PAF antagonism has been investigated for several diseases, with variable results thus far. Further investigation of its role in pathology and therapeutic modulation is highly anticipated because of the pressing need for more selective and targeted therapy for the management of severe anaphylaxis.

Anaphylaxis from bee pollen supplement

A 30-year-old woman was referred to the allergy clinic after having an anaphylactic reaction. Before the reaction, the patient had recently started taking three health supplements: bee pollen, omega 3-6-9 oil and vitamin D3. Ten minutes after she took her second dose of supplements, the patient’s

Assessment of ERCC1 and XPF Protein Expression Using Quantitative Immunohistochemistry in Nasopharyngeal Carcinoma Patients Undergoing Curative Intent Treatment

PURPOSE We sought to evaluate the prognostic/predictive value of ERCC1 and XPF in patients with nonmetastatic nasopharyngeal carcinoma (NPC) treated with curative intent. METHODS AND MATERIALS ERCC1 and XPF protein expression was evaluated by immunofluorescence combined with automated quantitative analysis (AQUA) using the FL297 and 3F2 antibodies, respectively. ERCC1 and XPF protein expression levels were correlated with clinical outcomes. RESULTS Patient characteristics were as follows: mean age 52 years (range, 18-85 years), 67% male, 72% Karnofsky performance status (KPS) ≥ 90%, World Health Organization (WHO) type 1/2/3 = 12%/28%/60%, stage III/IV 65%. With a median follow-up time of 50 months (range, 2.9 to 120 months), the 5-year overall survival (OS) was 70.8%. Median standardized nuclear AQUA scores were used as cutpoints for ERCC1 (n=138) and XPF (n=130) protein expression. Agreement between dichotomized ERCC1 and XPF scores was high at 79.4% (kappa = 0.587, P<.001). Neither biomarker predicted locoregional recurrence, DFS, or OS after adjustment for age and KPS, irrespective of stratification by stage, WHO type, or treatment. CONCLUSIONS Neither ERCC1 nor XPF, analyzed by quantitative immunohistochemistry using the FL297 and 3F2 antibodies, was prognostic or predictive in this cohort of NPC patients.

Dose delivery analysis of weekly versus 3-weekly cisplatin concurrent with radiation therapy for locally advanced nasopharyngeal carcinoma (NPC).

Objective:The purpose of this study was to analyze the dose delivery and toxicity of weekly cisplatin versus high-dose cisplatin given every 3 weeks in a tertiary oncology clinic. Methods:From January 2000 to July 2009, patients with biopsy-proven nasopharyngeal carcinoma receiving concurrent cisplatin with curative-intent radiation therapy (RT) were included. Before 2005, most patients received cisplatin (Q3) (100 mg/m2 intravenously days 1, 22, and 43 of RT) and 3-dimensional conformal RT (66 Gy, 33 fractions). After 2005, most patients received weekly cisplatin (Q1) (40 mg/m2 intravenously weekly for 7 wk of RT) and intensity-modulated radiotherapy (70 Gy, 35 fractions). Results:Seventy-three patients were analyzed: 45 for Q1 and 28 for Q3. Cumulative doses ≥200 mg/m2 were achieved in 80% of Q1 and 86% of Q3 patients, respectively. Dose reduction due to toxicity was required in 2/45 (4%) of Q1 patients compared with 11/28 (39%) of Q3 patients (P=0.0003). Toxicities in Q1 and Q3 patients included: hospitalization for acute toxicity in 20% and 35.7%; mean weight loss 10.85% and 8.75%; percutaneous endoscopic gastrostomy tube placement in 25.6% and 29.6%; and grade 3 dehydration in 11.1% and 17.9%, respectively. Median follow-up time was 3 years for Q1 and 6 years for Q3 patients. Median disease-free survival was 46 months for the Q1 group and 53 months for the Q3 group (P=0.667). There was no difference in overall survival between Q1 and Q3. Conclusions:In this series, weekly 40 mg/m2 cisplatin and 3-weekly 100 mg/m2 cisplatin showed similar deliverability, toxicity profiles, and outcomes. At our center, weekly cisplatin is standard of care for patients with locally advanced nasopharyngeal carcinoma undergoing chemoradiotherapy.

Oral allergy syndrome and risk of food-related anaphylaxis: a cross-sectional survey analysis

Background Oral Allergy Syndrome (OAS) is an IgE-mediated allergic response to fresh fruits, nuts and vegetables caused by cross-reactivity between pollen allergens and structurally similar food proteins. Alder pollen is a prominent allergen in coastal British Columbia, present at high levels from FebruaryApril. We hypothesized that this exposure may lead to increased prevalence of Alder pollen allergy and OAS. We sought to determine our population-based prevalence, cross-reactivity patterns, and incidence of food-related anaphylaxis.

ATM, THMS, and RRM1 protein expression in nasopharyngeal carcinomas treated with curative intent

In advanced nasopharyngeal carcinoma (NPC), biomarkers may help predict survival.

Acute generalized exanthematous pustulosis from imatinib

Background Acute generalized exanthematous pustulosis (AGEP) is rare cutaneous drug reaction involving drug-specific T lymphocytes and neutrophilic inflammation. Imatinib is a tyrosine kinase inhibitor used widely in treatment of chronic myeloid leukemia (CML). Rash is a common side effect of imatinib, occurring in up to 30% of patients. However, there are only two case reports to date describing AGEP secondary to imatinib.

Abstract 3545: ATM, TS and RRM1 protein expression in nasopharyngeal carcinomas (NPC) treated with curative intent.

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC Background: Although NPC is a radiation and chemotherapy sensitive tumour, some patients still relapse with distant metastatic disease. Biomarkers are needed to help identify which patients are at higher risk of relapse and/or who will respond to therapy. Ataxia telangiectasia mutated (ATM) is a putative marker for radiation sensitivity while, thymidylate synthetase (TS), and ribonucleotide reductase subunit M1 (RRM1) levels have been associated with response to 5FU and gemcitabine, two chemotherapy agents often used in NPC. We characterized the protein expression of ATM, TS, and RRM1 in tumours from 146 patients with non-metastatic NPC treated at two Canadian institutions between Jan 2000-Dec 2007 and explored the relationship of each marker with clinical outcomes. Methods: Pre-treatment, formalin-fixed, paraffin-embedded NPC tumour specimens were assembled in a tissue microarray. ATM, TS and RRM1 protein expression were evaluated by quantitative fluorescence immunohistochemistry and with automated quantitative digital image analysis (AQUA) using the Y170, TS106 and 60073-2-Ig monoclonal antibodies respectively. Protein expression levels were assessed in tumour, tumour cytoplasm, tumour nuclear, and non-malignant tumour stroma compartments. The effect of protein expression levels on overall survival (OS) and disease-free survival (DFS) was examined using Cox regression. For all biomarkers, AQUA scores were analyzed in quartiles. Results: Patient characteristics were: mean age=52 years (SD=12.2; range 18 to 85), 67% male, 73% KPS ≥ 90%, WHO type 1/2/3=11%/28%/61%, stage III/IV=65%. Fifty-eight (40%) patients received platinum-based CRT; the remainder was treated with RT alone. With a median follow-up of 50 months (range 3 to 120), the 5 year survival rates were 71% (95% CI=62%-78%) for OS and 48% (95% CI=39%-57%) for DFS. After adjusting for KPS, stage, and WHO type, OS was worse for the group with tumour:stromal (T/S) ATM ratios in the top quartile versus those in the lowest quartile (p=0.049). OS was also worse for tumours with TS levels in the lowest quartile versus the top quartile (p=0.033). RRM1 was not associated with outcomes (p=0.748). There was no significant effect of any of the biomarkers on disease-free survival (ATM T/S p=0.708; TS p=0.978; RRM1 p=0.918). Conclusions: In our cohort of non-metastatic NPC patients, relative overexpression of ATM and low TS protein levels were associated with worse outcomes suggesting further investigation of these potential biomarkers is warranted. Citation Format: Alexander C. Klimowicz, Amanda Jagdis, Tien Phan, Janessa J. Laskin, Harold Y. Lau, Jodi E. Siever, Stephanie K. Petrillo, Thomas A. Thomson, M. Sarah Rose, Anthony M. Magliocco, Desiree Hao. ATM, TS and RRM1 protein expression in nasopharyngeal carcinomas (NPC) treated with curative intent. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3545. doi:10.1158/1538-7445.AM2013-3545