2008 Summer Meeting - Leeds - The Pathological Society of Great ...

P159Immunohistochemistry Biomarker Validation for Use inClinical Trials – Case Study with Src Inhibitor AZD0530SR Morgan 1 ,CWomack 1 ,VJacobs 1 , TP Green 1 ,MStuart 11 AstraZeneca, Alderley Park, Macclesfield, Cheshire, UKBiomarkers of Src and small-molecule drug inhibitory activity have beencritical in supporting early clinical development of AZD0530. Wedescribe preclinical development and clinical implementation of tumour tissuebiomarker assays of Src activity utilizing immunohistochemistry (IHC)endpoints.Immunohistochemical assays utilizing phosphospecific antibodies to integrinlinkedSrc substrates focal adhesion kinase (FAK) and paxillin were developedin the laboratory using AZD0530-treated cell lines and xenograft models.Biopsies were used to develop assay protocols in a clinical setting beforeimplementation in a Phase I study in patients with advanced solid malignancies.IHC was performed on formalin fixed, paraffin embedded tissue samples frompatients where pre- and post-treatment tumour biopsies were collected.In this Phase I study, AZD0530 reduced the levels of p-FAK and p-paxillin insome but not all patients. Biomarker changes were seen at all doses and in alltumour types. Paired pre- and post-treatment biopsies were evaluable forblinded IHC analysis in 26 and 22 out of 51 patients for p-FAK and p-paxillin,respectively. Concordance was significant for both biomarkers: 21/26 samples(81%) were correctly assigned based on p-FAK staining (P=0.001, 80% exactCI: 0.672–0.899) and 15/22 (68%) based on p-paxillin staining (P=0.067, 80%exact CI: 0.523–0.813).These data describe the validation of IHC biomarkers to demonstrate in earlyclinical development the drug inhibition of Src pathway signalling, as proof ofmechanism for AZD0530 in target tissues of cancer patients.P161Differential expression of microRNA92 and one of its genetargets, ER beta, in normal and malignant breast tissues andcell linesH Al-Nakhle 1 , PA Burns 1 ,MCummings 1 , TA Hughes 1 ,AM Shaaban 1 ,VSpeirs 11 Leeds Institute of Molecular Medicine, University of LeedsER expression in breast cancer is lower than in normal breast, suggesting ERis downregulated during carcinogenesis. Silencing of ER in breast tumoursmay be mediated by one or more specific microRNAs. The aim of this studywas to identify expression profiles of ER mRNA and to correlate this withmiR92, a predicted ER target, in normal and malignant breast tissues and celllines. Interrogation of the Sanger miRBase database revealed a conservedtarget-site for miR92 within the ER 3'-UTR at 955– 976 nt, confirming itspotential role as a regulator of ER. Total RNA was extracted from breasttumours (n=10), paired normal/ tumour samples (n=11) and breast cancer celllines. TaqMan® MicroRNA Assay hsamiR92 was used to estimate miR92expression and SYBR green QRT-PCR was used to detect ER expression inall samples. We observed that miR92 was differentially expressed in normalbreast tissues and cancers. Breast tumours with abundant miR92 expressiondisplayed a significant decrease in ER expression (n=14; p=0.005). Thereverse was observed in tumours with low miR92 expression (n=7; p=0.01). Inbreast cancer cell lines with low ER expression (BT20 and MDAMB453),miR92 expression was also high. In addition, there was a statistically significantdifference in ER expression between normal and breast tumours (p=0.004) butnot miR92.Our results suggest that our observed downregulation of ERexpression in breast tumours and cell lines may be mediated by miR92. Currentwork is confirming this.P160Thymoma associated with hypergammaglobulinemia: A casereportHR Suzangar 1 ,MSuzangar 2 ,MHaghighi 3 , D Taheri 1 , R Sherkat 41 Dept of Pathology, Isfahan University of Medical Sciences, Isfahan, Iran2 MRCS Manchester Royal Infirmary, Manchester, UK. 3 Dept of Dentistry,Khorasgan Azad University, Isfahan, Iran. 4 Dept of Infectious Diseases,Isfahan University of Medical Sciences, Isfahan, IranThymoma is a term that should be restricted to neoplasms of thymic epithelialcells, irrespective of the presence or the number of lymphocytes.The usual location of thymoma is the antrosuperior mediastinum; however ,this tumour can also occur in other mediastinal compartments, in the neck ,within the thyroid gland, in the pericardial cavity , the pulmonary hilum , withinthe lung parenchyma, or the pleura itself.The Association of thymoma withhypogammaglobulinemia,Mucocutaneous candidiasis and SLE is well known.Here we present a patient admitted in our hospital with buttock rashes regardedto be zoster of the buttock.In this past medical history he had recurrent cutaneous infections, lupuserythematosus , hypergammaglobulinemia, recurrent oral candidiasis and B-cell dysfunction.During the patient evaluation , a mass was found in the mediastinum andpercutaneous niddle biopsy confirmed the diagnosis of thymoma(type B3).This is the second case reported having thymoma in association withhypergammaglobulinema.Key word: Thymoma- hypergammaglobulinema- systemic lupus erythematosusP162EGFR, Her-2/neu and Cyclin D1 Gene Copy Number andProtein Expression in Head and Neck Squamous CellCarcinomaN Gaber 1 ,AS Jones 1 , TR Helliwell 11 University of LiverpoolThis study has examined the associations between, and prognostic value of,changes in chromosome number, gene copy number and protein expression ofEGFR, HER2/neu and cyclin-D1 by fluorescence in situ hybridization (FISH)and immunocytochemistry on tissue microarray sections of 200 cases of headand neck squamous cell carcinoma (HNSCC). Gene amplification was regardedas a ratio of 2 or more between the signals for the genes and the chromosomalcentromeres.Amplification of the cyclin-D1 gene was present in 31.3% cases withchromosome 11 trisomy in 41.9% cases and polysomy in 22.2% cases.Amplification correlated with high (>median) protein expression (p