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 Blood, 15 October 2008, Vol. 112, No. 8, pp. 3425-3433.
Prepublished online as a Blood First Edition Paper on June 10, 2008; DOI 10.1182/blood-2008-02-137372.

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NEOPLASIA

Gene expression predicts overall survival in paraffin-embedded tissues of diffuse large B-cell lymphoma treated with R-CHOP

Lisa M. Rimsza1, Michael L. LeBlanc2, Joseph M. Unger2, Thomas P. Miller3, Thomas M. Grogan1, Daniel O. Persky3, Ralph R. Martel4, Constantine M. Sabalos4, Bruce Seligmann4, Rita M. Braziel5, Elias Campo6, Andreas Rosenwald7, Joseph M. Connors8, Laurie H. Sehn8, Nathalie Johnson9, and Randy D. Gascoyne9

1 Department of Pathology, University of Arizona, Tucson; 2 Fred Hutchison Cancer Research Center, Seattle, WA; 3 Arizona Cancer Center, Tucson; 4 High Throughput Genomics, Tucson, AZ; 5 Department of Pathology, Oregon Health Sciences Center, Portland; 6 Department of Pathology, University of Barcelona, Barcelona, Spain; 7 Department of Pathology, University of Würzburg, Würzburg, Germany; and Departments of8 Medical Oncology and 9 Pathology, British Columbia Cancer Agency, Vancouver, BC

Gene expression profiling (GEP) on frozen tissues has identified genes predicting outcome in patients with diffuse large B-cell lymphoma (DLBCL). Confirmation of results in current patients is limited by availability of frozen samples and addition of monoclonal antibodies to treatment regimens. We used a quantitative nuclease protection assay (qNPA) to analyze formalin-fixed, paraffin-embedded tissue blocks for 36 previously identified genes (N = 209, 93 chemotherapy; 116 rituximab + chemotherapy). By qNPA, 208 cases were successfully analyzed (99.5%). In addition, 15 of 36 and 11 of 36 genes, representing each functional group previously identified by GEP, were associated with survival (P < .05) in the 2 treatment groups, respectively. In addition, 30 of 36 hazard ratios of death trended in the same direction versus the original studies. Multivariate and variable cut-off point analysis identified low levels of HLA-DRB (< 20%) and high levels of MYC (> 80%) as independent indicators of survival, together distinguishing cases with the worst prognosis. Our results solve a clinical research problem by demonstrating that prognostic genes can be meaningfully quantified using qNPA technology on formalin-fixed, paraffin-embedded tissues; previous GEP findings in DLBCL are relevant with current treatments; and 2 genes, representing immune escape and proliferation, are the common features of the most aggressive DLBCL.


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