Aims The cell of origin (COO) based molecular characterisation into germinal centre B-cell-like (GCB) and activated B-cell-like (ABC) subtypes are central to the pathogenesis and clinical course in diffuse large B-cell lymphoma (DLBCL). Globally, clinical laboratories employ pragmatic but less than ideal immunohistochemical (IHC) assay for COO classification. Novel RNA-based platforms using routine pathology samples are emerging as new gold standard and offer unique opportunities for assay standardisation for laboratories across the world. We evaluated our IHC protocols against RNA-based technologies to determine concordance; additionally, we gauged the impact of preanalytical variation on the performance of Lymph2Cx assay.
Methods Diagnostic biopsies (n=104) were examined for COO classification, employing automated RNA digital quantification assay (Lymph2Cx). Results were equated against IHC-based COO categorisation. Assay performance was assessed through its impact on overall survival (OS).
Results 96 (92%) informative samples were labelled as GCB (38/96; 40%) and non-GCB (58/96; 60%) by IHC evaluation. Lymph2Cx catalogued 36/96 (37%) samples as GCB, 45/96 (47%) as ABC and 15/96 (16%) as unclassified. Lymph2Cx being reference, IHC protocol revealed sensitivity of 81% for ABC and 75% for GCB categorisation and positive predictive value of 81% versus 82%, respectively. Lymph2Cx-based COO classification performed superior to Hans algorithm in predicting OS (log rank test, p=0.017 vs p=0.212).
Conclusions Our report show that current IHC-based protocols for COO classification of DLBCL at UKM Malaysia are in line with previously reported results and marked variation in preanalytical factors do not critically impact Lymph2Cx assay quality.
- Diffuse large B-cell lymphoma
- Cell of origin
- DLBCL -GCB subtype
- DLBCL -ACB subtype
- Hans algorithm.
Statistics from Altmetric.com
Handling editor Mary Frances McMullin.
Contributors KCP, NM, AA and AM conceived and performed the study, compile/analyse the data and wrote the manuscript. FAR, NMST, M-TS-R, RZA and GE collected samples, performed analysis and reviewed the manuscript.
Funding K-CP, NMST, FAR and NM were supported by UKM grant no (UKM220.127.116.11/244/DIP-2012- 10/3) while AA was supported by research grant from Alberta Cancer Foundation (grant # 25999).
Competing interests None declared.
Ethics approval UKM Ethics Committee (UKM18.104.22.168/244/DIP-2012-10/3 dated 31 Oct 2012).
Provenance and peer review Not commissioned; externally peer reviewed.