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Prediction of patient survival by multiplexed quantitative (AQUA™) analysis of tissue biomarkers in primary cutaneous melanoma
Aaron J Berger, Robert L. Camp, Gretchen Graff, Harriet Kluger, Stephan Ariyan, David L. Rimm
Yale University School of Medicine, New Haven, CT
Objective: Develop an assay that will predict disease-specific survival by multiplexed analysis of in situ protein expression in primary melanoma.
Design / Setting: A tissue microarray (TMA) was constructed with specimens from the Yale Department of Pathology: 214 primary and 293 metastatic melanomas. AQUA (determines exact expression levels of protein within overall and subcellular compartments) was used to assess association of 48 biomarkers with melanoma-specific survival. Genetic algorithms were instituted to determine the optimal number of biomarkers.
Patients: Serial collection of melanoma specimens resected between 1959 and 1994 from the Yale University Department of Pathology Archives resulted in 507 patients.
Main Outcome Measure: Melanoma-specific survival.
Results: Sixty-six variables (derived from 48 protein assays) were analyzed; thirteen are significantly (p<0.1) associated with survival by Cox univariate analysis. Genetic algorithms were run to convergence (>20X106 generations), resulting in the same five-member solution: ATF2_cytoplasm by nuclear; nuclear p21; p16_cytoplasm by nuclear; Beta catenin; Fibronectin. Addition of the genetic algorithm to Breslow depth significantly aids in the stratification of patients.
Conclusion: It is possible to improve upon the best prognostic index in primary melanoma (Breslow depth) by using in situ measurement of protein expression in primary melanoma. We aim to identify those patients who are likely to develop metastatic disease, regardless of Breslow depth.
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