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Purpose. To determine the presence of genetic heterogeneity (for chromosomes 3 and 8) in uveal tract melanoma and correlate this with tumour size and morphology. Methods. Ninety-five patients who underwent enucleation for primary uveal melanoma in the period of 2009-2011 were included in the study. Tumour dimensions were preoperatively measured (height, maximal diameter). Corresponding areas and volumes were calculated. Tumour morphology was classified in separate subcategories (collar-stud, dome-shaped, bilobed, multilobed, diffuse and other) through B-mode ultrasonography. Double blind analysis of uveal tract melanoma by fine needle aspiration biopsy (FNAB) (from the centre of the tumour) vs punch biopsy (from the base of the tumour) to assess the presence of genetic heterogeneity of chromosomes 3 and 8 using the appropriate probes. Results were correlated with tumour dimensions, tumour size according to COMS classification and tumour morphology. In addition the yield of FNAB was determined and similar correlations were done. Results. Fine needle aspiration biopsy yield i.e. sufficient cells for diagnosis was seen in 77% (73/ 95) of tumours. Yield was higher in large tumours according to the COMS classification (83% of large melanomas). The yield was worst for bilobed tumours (36%, 4/11). Genetic heterogeneity (GH) between FNAB and punch biopsies was assessed in the remaining 72 patients. GH was 4% (3 out of 72 tumours) for chromosome 3 and 21% (15 out of 72 tumours) for chromosome 8. Genetic heterogeneity strongly correlated with collar-stud morphology for chromosome 3 (100%, 3 out of 3 tumours) and for chromosome 8 (53%, 8 out of 15 tumours). In addition heterogeneity for chromosome 8 strongly correlated with large melanomas, according to COMS classification (73%, 11 out of 15 tumours) Conclusions. Genetic heterogeneity for chromosomes 3 and 8 is strongly associated with collar-stud lesion morphology. Heterogeneity for chromosome 8 is correlated with large uveal melanoma. Financial disclosure. None 49 UM8 DEVELOPMENTS IN PREDICTING METASTASIS FROM CHOROIDAL MELANOMA B.E. Damato, A. Eleuteri, A.F. Taktak, S.E. Coupland (bertil.damato@ gmail.com) Royal Liverpool University Hospital; University of Liverpool Purpose. We previously developed prognostic models using neural networks but these were inaccurate when baseline data were incomplete. We therefore devoped new models based on Accelerated Failure Time. The aim of this presentation is to evaluate our improved tool in terms of its ability to estimate survival probability after treatment of choroidal melanoma. Methods. Data from 3653 patients were used to generate models for predicting all-cause mortality. The program produced all-cause mortality curves for patients and controls, thereby allowing risk of metastatic mortality to be estimated. Results. The predicted morality correlated well with the observed mortality (Kolmogorov-Smirnov statistic, 0.79. p value 0.80). The C-index for discrimination (correlating predictions with risk factors) was 0.75 for the clinical model and 0.79 for the laboratory model (with fewer data). Conclusions. Our AFT model is a significant improvement over neural networks and provides reasonably reliable prognosis relevant to individual patients with choroidal melanoma. Financial disclosure. None UVEAL MELANOMA Abstracts 98 444 UM9 FISH-BASED PROGNOSTICATION OF UVEAL MELA- NOMA M. Turell 1, R. Tubbs 2 , C. Biscotti 3 , L. Schoenfield 3, Y. Sun 2 , G. Bebek 4 , Y. Saunthararajah 5, P. Triozzi 6, A. Singh1 (turellm2@ccf.org) 1. Cole Eye Institute, Cleveland Clinic Foundation 2. Department of Molecular Pathology, Cleveland Clinic Foundation 3. Department of Anatomic Pathology, Cleveland Clinic Foundation 4. Center for Proteomics and Bioinformatics, Case Western Reserve University 5. Hematologic Oncology & Blood Disorders, Cleveland Clinic Foundation 6. Solid Tumor Oncology, Cleveland Clinic Foundation, Cleveland, OH Purpose. To compare detection of monosomy 3 in uveal melanoma using single nucleotide polymorphism array (SNP-A) and fluorescence in situ hybridization (FISH) Methods. Consecutive cases of uveal melanoma treated by enucleation from 2004 to 2010 were analyzed. DNA was isolated from fresh frozen tissue and was analyzed by SNP-A (Illumina, San Diego, CA) and by FISH using both centromeric (CEP3) and locus-specific (3p26) probes (Abbott Molecular, Des Plains, IL). A total of 200 interphase cells were scored and a cut-point of 20% was used to determine monosomy 3 status. Results. In this series, there were 50 Caucasian patients including 28 males (56%) and 22 females (44%) with an average age at diagnosis of 64 years. Tumor location was choroidal in 24 (48%), ciliochoroidal in 18 (36%), and iridociliochoroidal in 8 cases (16%). Monosomy 3 was detected by either SNP-A or FISH (CEP3) in 33 cases (66%). SNP-A detected monosomy 3 in 31 tumors (94%). CEP3 probe detected 25 cases of monosomy 3 (76%). Locus-specific FISH probe (3p26) detected deletion in 28 of 31 cases (90%) of monosomy 3 positive tumors by SNP-A and every case of of monosomy 3 by CEP3 probe. Conclusions. There is a strong correlation between monosomy 3 detected by SNP-A and by 3p26 deletion detected by locus-specific FISH probe. Financial disclosure. This work was supported by a Falk Trust grant and a Research to Prevent Blindness Challenge Grant, Department of Ophthalmology, Cleveland Clinic Lerner College of Medicine. 1819 UM10 FLUORESCENCE IN-SITU HYBRIDIZATION VS MULTI- PLEX LIGATION-DEPENDENT PROBE AMPLIFICATION FOR UVEAL MELANOMA PROGNOSTICATION: IN-VI- VO COMPARATIVE RESULTS Raffaele Parrozzani1, Elisabetta Pilotto2, Alessia Dario2, Giacomo Miglionico2, Edoardo Midena1,2 (parrozzani@libero.it) 1. GB Bietti Eye Foundation, IRCCS, Roma, Italy. 2. Department of Ophthalmology, University of Padova, Padova, Italy. Purpose. To compare fluorescence in-situ hybridization (FISH) and multiplex ligation-dependent probe amplification (MLPA) for uveal melanoma prognostication. Methods. Twenty-four patients affected by posterior uveal melanoma and scheduled for I125 brachytherapy were included in this prospective study. Patients underwent in-vivo 25-G transcleral FNAB (two passes) just before applying the radioactive plaque. Sampled material underwent both FISH and MLPA analysis using standard procedures. Follow-up was longer than 24 months.
Results. Follow-up was 31±8 months (range, 25-42 months). FISH analysis revealed monosomy 3 in twelve cases (50%). MLPA revealed monosomy 3 in thirteen cases (54%) and a 3p14-q29 deletion in one case (4%) (classified as monosomy 3 by FISH). Nine patients (41%) developed metastatic disease during follow-up, including the case showing monosomy 3 only by MLPA. Patient with partial chromosome 3 deletion is still alive without metastases. Conclusions. MLPA allows to obtain more information than standard FISH in uveal melanoma prognostication. The biological and prognostic value of partial chromosome 3 deletion, as well as others subtle chromosomes alterations or complex MLPA results, remains unclear. Financial disclosure. None 2331 UM11 UVEAL MELANOMA: AN ANALYSIS OF CELLULAR FEATURES AND COMPARISON TO MONOSOMY 3 STATUS C. V. Biscotti1, A Lott-Limbach1, R. R. Tubbs2, M. E. Turell3, Y Sun2, A. D. Singh3 (biscotc@ccf.org) 1. Departments of Anatomic Pathology, 2. Molecular Pathology and 3. Ophthalmology, Cleveland Clinic, Cleveland, Ohio USA Purpose. Uveal melanoma patients segregate into two prognostic groups. Approximately half will eventually manifest metastases. Unfortunately, most experts agree that the die is cast by the time of diagnosis. We analyzed a series of uveal melanoma FNA samples to describe the cellular features and correlate these with the results of FISH analysis for monosomy 3. Methods. All patients had a clinical diagnosis of uveal melanoma based on history and physical examination including ophthalmoscopic examination performed by one of the authors (ADS).The patients consented to this study which was funded by the Falk Trust. FNA using a 25G needle was performed at the time of radiation therapy plaque placement or enucleation/excision by two of the authors (ADS and MET). In vivo samples were obtained by transvitreal or transscleral approach depending on tumour location. Aspirate samples were entirely rinsed into 2-3 ml of normal saline, visually examined for adequacy, and then entirely transferred to a cell preservative, CytoLyt (Hologic Corp, Marborough MA). ThinPrep® processing yielded 4 alcohol fixed Papanicolaou stained slides per case. One of the authors (CVB) analyzed the slides for cellular features including cell type (pure epithelioid, pure spindle, or mixed), nuclear grade (1, 2, or 3), and the presence or absence of necrosis, tumour infiltrating lymphocytes (Tils), and melanin. Slides with well preserved melanoma cells in sufficient numbers were destained and analyzed for monosomy 3 by FISH. An adequate FISH study required 200 cells. A positive result required monosomy 3 in at least 20% of cells. Results. Ninety patients with a clinical diagnosis of uveal melanoma consented to the study. This included 45 men and 45 women with ages ranging from 26 to 96 years (mean 62). Tumours involved the choroid 61(68%) cases, ciliochoroid 21(23%) cases, iridociliary 5(6%) cases and iris 3(3%) cases and ranged from 3 x 3 mm to 20 x 24 mm in basal dimensions and 1.2 mm to 15 mm in height. Fifty-eight (64%) patients had in vivo FNA either transvitreal 33 (57%) or transscleral 25 (43%).While thirty-two (36%) patients had FNA of enucleation/excision specimens. FNA samples had diagnostic melanoma cells in 80 (89%) cases including 68 (76%) with a 200 cell FISH count. Six patients did not have data recorded for the specific cellular features. This yielded 62 patients for the cytology-monosomy 3 correlation. Tumour cell type UVEAL MELANOMA Abstracts 99 included 40 (65%) mixed and 22(35%) spindle. There were no pure epithelioid tumours. Most (40 cases, 65%) tumours had grade 2 nuclei. Grade 3 and grade 1 nuclei occurred in only 13(21%) and 9(15%) tumors, respectively. Tils occurred in 18 (29%) tumours. FISH analysis identified monosomy 3 in 28(45%) tumours. Monosomy 3 occurred significantly more often in tumours with grade 3 nuclei (69% versus 39% in tumours with grade 1 or 2 nuclei, p=0.0498). Monosomy 3 occurred more often in tumours with TILs (61% versus 39% in tumours lacking TILs); this difference is not significant (p=0.11). Conclusions In our series most uveal melanomas had a relatively consistent cellular appearance characterized by at least some spindle cells, no more than moderate nuclear atypia, and melanin. In our experience monosomy 3 occurred more often in tumours with grade 3 nuclei or Tils; however, the positive predictive value of these cellular features is low, limiting clinical utility. Financial disclosure. None 2321 UM12 RETROSPECTIVE ANALYSIS OF 700 FINE-NEEDLE ASPIRATION BIOPSIES OF SOLID INTRAOCULAR TUMORS: CHANGING INDICATIONS DURING LONG- TERM EXPERIENCE Zélia M. Corrêa, James J. Augsburger correazm@uc.edu Department of Ophthalmology, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA Purpose. Describe the changing indications for and techniques of FNAB of intraocular tumours in a single group’s experience during a period of over 30 years. Methods. Retrospective records review and data analysis on 700 cases of FNAB of a solid intraocular tumour performed by the authors during the period 9/1980 through 6/2011. Results. During the first 5 years, most FNABs were investigational (including post-enucleation & post-resection to evaluated different calibers and lengths needles, different methods of aspiration, routes of needle passage, cellular yield, specimen processing, cytologichistologic correlation, and provide experience with cytology of the various types of tumours to the pathologists reading these samples). All clinical FNABs (diagnostic or confirmatory) during this period were performed under an IRB approved protocol in which clinical diagnostic accuracy, cytopathologic diagnostic accuracy, complications of FNAB, and percentage of cases in which the results of FNAB changed patient management were evaluated. In 1985, the IRB determined that FNAB of intraocular tumours by us no longer needed to be regarded as investigational. During the ensuing 10 to 15 years, most FNABs we performed were clinical diagnostic or confirmatory biopsies. Initially, amelanotic uveal melanoma versus metastatic cancer to uvea was the most common differential diagnosis, and later on, it became large uveal nevus versus small uveal melanoma. Our practice relocated to a different institution in 1999 prompting a new round of post-enucleation FNABs to familiarize our new pathologists with the cytopathologic features of intraocular tumours. In 2006, retrospective analysis of 25-year experience with FNAB of clinically diagnosed melanocytic uveal tumours showed that cytologic classification to be an independently significant prognostic factor for metastasis and metastatic death. From that point on, all of our FNABs on clinically diagnosed melanocytic uveal tumours have been regarded as prognostic. In 2007, we began to collaborate in a prospective IRB-approved multicenter (COOG) study of the prognostic significance of
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List of Congress Meetings of the In
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Table of Contents Welcome address K
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Keynote Lectures Lymphoma of the oc
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The venue for the Biannual Meeting
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Buenos Aires Downtown MAP 12 Venue
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MORNING 8.30-10.10 Papers (Moderato
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50 RES 26 USING THE GLYCOLYTIC INHI
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a reduction in the number of living
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1909 Rb14 RESULTS OF PATIENTS WITH
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Posters Retinoblastoma 2006 RBp100
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120 RBp135 MANAGEMENT OF AN ECTOPIC
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After treatment, 10 patients lived
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Methods. Six cycles of carboplatin
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1940 RB15 PROTON THERAPY FOR RECURR
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4. Fluoroscopy for cannula placemen
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Methods. A retrospective chart revi
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Ruth A. Kleinerman1, Chu-ling Yu1,
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