title: Whole genome expression profiling based on paraffin embedded tissue can be used to classify diffuse large B-cell lymphoma and predict clinical outcome.
abstract: This study tested the validity of whole-genome expression profiling (GEP) using RNA from formalin-fixed, paraffin-embedded (FFPE) tissue to sub-classify Diffuse Large B-cell Lymphoma (DLBCL), in a population based cohort of 172 patients. GEP was performed using Illumina Whole Genome cDNA-mediated Annealing, Selection, extension & Ligation, and tumours were classified into germinal centre (GCB), activated B-cell (ABC) and Type-III subtypes. The method was highly reproducible and reliably classified cell lines of known phenotype. GCB and ABC subtypes were each characterized by unique gene expression signatures consistent with previously published data. A significant relationship between subtype and survival was observed, with ABC having the worst clinical outcome and in a multivariate survival model only age and GEP class remained significant. This effect was not seen when tumours were classified by immunohistochemistry. There was a significant association between age and subtype (mean ages ABC - 72·8 years, GC - 68·4 years, Type-III - 64·5 years). Older patients with ABC subtype were also over-represented in patients who died soon after diagnosis. The relationship between prognosis and subtype improved when only patients assigned to the three categories with the highest level of confidence were analysed. This study demonstrates that GEP-based classification of DLBCL can be applied to RNA extracted from routine FFPE samples and has potential for use in stratified medicine trials and clinical practice.
authors.affiliation: Haematological Malignancy Diagnostic Service, St. James's Institute of Oncology, Leeds, UK. sharonb@hmds.org.uk
authors.forename: Sharon L
authors.initials: SL
authors.lastname: Barrans
authors.forename: Simon
authors.initials: S
authors.lastname: Crouch
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Care
authors.forename: Lisa
authors.initials: L
authors.lastname: Worrillow
authors.forename: Alex
authors.initials: A
authors.lastname: Smith
authors.forename: Russell
authors.initials: R
authors.lastname: Patmore
authors.forename: David R
authors.initials: DR
authors.lastname: Westhead
authors.forename: Reuben
authors.initials: R
authors.lastname: Tooze
authors.forename: Eve
authors.initials: E
authors.lastname: Roman
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Jack
mesh_terms: D000368:Aged; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D015894:Genome, Human; D055106:Genome-Wide Association Study; D006801:Humans; D007150:Immunohistochemistry; D016403:Lymphoma, Large B-Cell, Diffuse / Q000145:classification* / Q000235:genetics / Q000473:pathology; D008297:Male; D008875:Middle Aged; D016612:Paraffin Embedding; D011379:Prognosis; D016896:Treatment Outcome