title: Microarray gene expression analysis of fixed archival tissue permits molecular classification and identification of potential therapeutic targets in diffuse large B-cell lymphoma.
abstract: Refractory/relapsed diffuse large B-cell lymphoma (DLBCL) has a poor prognosis. Novel drugs targeting the constitutively activated NF-κB pathway characteristic of ABC-DLBCL are promising, but evaluation depends on accurate activated B cell-like (ABC)/germinal center B cell-like (GCB) molecular classification. This is traditionally performed on gene microarray expression profiles of fresh biopsies, which are not routinely collected, or by immunohistochemistry on formalin-fixed, paraffin-embedded (FFPE) tissue, which lacks reproducibility and classification accuracy. We explored the possibility of using routine archival FFPE tissue for gene microarray applications. We examined Affymetrix HG U133 Plus 2.0 gene expression profiles from paired archival FFPE and fresh-frozen tissues of 40 ABC/GCB-classified DLBCL cases to compare classification accuracy and test the potential for this approach to aid the discovery of therapeutic targets and disease classifiers in DLBCL. Unsupervised hierarchical clustering of unselected present probe sets distinguished ABC/GCB in FFPE with remarkable accuracy, and a Bayesian classifier correctly assigned 32 of 36 cases with >90% probability. Enrichment for NF-κB genes was appropriately seen in ABC-DLBCL FFPE tissues. The top discriminatory genes expressed in FFPE separated cases with high statistical significance and contained novel biology with potential therapeutic insights, warranting further investigation. These results support a growing understanding that archival FFPE tissues can be used in microarray experiments aimed at molecular classification, prognostic biomarker discovery, and molecular exploration of rare diseases.
authors.affiliation: The University of Manchester, Manchester Cancer Research Centre, Manchester, United Kingdom. kim.linton@manchester.ac.uk
authors.forename: Kim
authors.initials: K
authors.lastname: Linton
authors.forename: Christopher
authors.initials: C
authors.lastname: Howarth
authors.forename: Mark
authors.initials: M
authors.lastname: Wappett
authors.forename: Gillian
authors.initials: G
authors.lastname: Newton
authors.forename: Cynthia
authors.initials: C
authors.lastname: Lachel
authors.forename: Javeed
authors.initials: J
authors.lastname: Iqbal
authors.forename: Stuart
authors.initials: S
authors.lastname: Pepper
authors.forename: Richard
authors.initials: R
authors.lastname: Byers
authors.forename: Wing John
authors.initials: WJ
authors.lastname: Chan
authors.forename: John
authors.initials: J
authors.lastname: Radford
mesh_terms: D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D016403:Lymphoma, Large B-Cell, Diffuse / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D058990:Molecular Targeted Therapy; D016328:NF-kappa B / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016612:Paraffin Embedding; D016707:Tissue Fixation
chemical_list: D016328:NF-kappa B